I

m

T ('

v v

A MANUAL
of the

ASPERGILLI

By

CHARLES THOM

Collaborator, Northern Regional Research Laboratory,

Formerly Principal Mycologist, Bureau of

Plant Industry, U. S. Department of

Agriculture, Washington, D. C.

and
KENNETH B. RAPER

Senior Microbiologist. Fermentation Division, Northern Regional

Research Laboratory, Bureau of Agricultural and

Industrial Chemistry, U. S. Department of

Agriculture, Peoria, Illinois

BALTIMORE

THE WILLIAMS & WILKINS COMPANY

1945

Copyright, 1945
The Williams & Wilkins Company

Made in the United Stales of America

Published May, 1945
Reprinted January, 1951

Composed and Printed at the

WAVERLY PRESS, INC.

for

THE WILLIAMS & WILKINS COMPANY

Baltimore, Md., U. S. A.

CONTENTS

Preface vii

Part I. General Discussion 1

Chapter I. Historical Introduction 3

Chapter II. Classification, Generic Diagnosis, and Synonymy 6

Chapter III. Morphology and Description 10

Chapter IV. Cultivation and Examination 31

Chapter V. Preservation of Cultures 50

Chapter VI. Variation 63

Part II. The Manual Proper 79

Chapter VII. The Use of the Manual 81

Chapter VIII. The Aspergillus clavatus Group 92

Chapter IX. The Aspergillus glaucus Group 100

Chapter X. The Aspergillus fumigatus Group 148

Chapter XI. The Aspergillus nidulans Group 155

Chapter XII. The Aspergillus ustus Group 171

Chapter XIII. The Aspergillus flavipes Group 179

Chapter XIV. The Aspergillus versicolor Group 183

Chapter XV. The Aspergillus terreus Group 195

Chapter XVI. The Aspergillus candidus Group 206

Chapter XVII. The Aspergillus niger Group 214

Chapter XVIII. The Aspergillus wentii Group 241

Chapter XIX. The Aspergillus tamarii Group 250

Chapter XX. The Aspergillus fiavus-oryzae Group 259

Chapter XXI. The Aspergillus ochraceus Group 273

Part III. Reference Material 287

Chapter XXIT. Topical Bibliography 289

Chapter XXIII. General Bibliography 319

Chapter XXIV. Check List of Species and Genera 331

Chapter XXV. Accepted Species, Varieties, and Mutations 360

Index 363

PREFACE

Aspergillus as the name for a genus of molds dates back to Micheli
(1729), but it was not until the middle of the 19th Century that the Asper-
gilli began to be recognized as active agents in many decay processes, as
occasional causes of human and animal disease, and as fermenting agents
capable of producing valuable biochemical products. Various taxonomic
efforts were made. DeBary, Fresenius, van Tieghem, and others described
the particular species that they used. Bainier described in brief inadequate
terms all the Aspergilli he found. Wilhelm and Wehmer reviewed the
literature, described and figured the forms they knew; these were few in
number, but the work was done so well that it fixed group types. In
1926, Thorn and Churchbrought all of this material together in amonograph.
The increased study devoted to the Aspergilli in recent years shows some
of their groupings to be inadequate. In addition, a large amount of new
material has accumulated. The Aspergilli have become increasingly
important as responsible agents in a number of industrial fermentations.
Many of them are being found capable of producing antibiotic substances
and their possible use in this field will undoubtedly be exhaustively ex-
plored. For these reasons, the need for a manual for those who wish to
identify Aspergilli under observation, without regard to the historical
aspects of the group, has become increasingly apparent.

This book is definitely a manual, not a monograph. It is based upon
comparative study of thousands of strains of Aspergilli in culture. Repre-
sentative strains giving the range of morphology and biochemical activity
in each species are maintained in the permanent collection of the Northern
Regional Research Laboratory. Consistent efforts have been made to
obtain the organisms actually used by authors who have put forward new
nomenclature. The manual thus seeks to present under species names only
living cultures known to the authors, although it seemed advisable to make
a few additions based upon literature. The species included are arranged
as far as possible into natural groups which bring together aggregates of
strains or species agreeing in important morphological characters. For
the most part, physiological or biochemical information, if available, in-
dicates related activities within these groups. The names selected for use
appear to be taxonomically correct. A large number of names are neces-
sarily rejected. If known to belong to some unidentifiable member of a
group, or believed from literature to be correctly placed there, each of
these names is accounted for in the discussion of the group. If the in-
formation available does not justify allocation to some species or species

vii

Vlll A MANUAL OF THE ASPERGILLI

aggregate, the name will be found in the check list with any information
at hand. Large gaps in our information about the Aspergilli still exist.
Some of these are pointed out in the text. The great activity of the
present day will undoubtedly render any arrangement of the Aspergilli
obsolete sooner or later, but it is believed that the classification put forward
here is at least temporarily practical.

Recognizing that any species name for an Aspergillus appearing at any
time in the literature may at some future time become important for
some unanticipated reason, an alphabetical check list giving each of the
names found, the author, the date, and the place of publication has been
included. An index reference to page in the manual, or the method of
disposal of the name, is added to bring the material to its greatest usefulness
as a ready reference.

Two types of bibliography are presented: a general bibliography,
alphabetical to author's name and sub-indexed as to date of publication
when necessary, includes authors of species and other investigators whose
work is cited in the text. In addition, a topical bibliography is presented.
Although incomplete, it is hoped that the latter will assist greatly in the
search for special literature on particular subjects. Believing that the
more recent literature on these subjects will generally be of the greatest
interest and value, the material is presented chronologically. Duplication
between the two bibliographies may or may not occur.

A manual, if it is to facilitate the identification of these molds by the
actual worker in the laboratory, must present descriptive and illustrative
material in as simple form as seems consistent with sound scholarship.
From our present point of view, the describer of a mold must know that
mold in fruiting form under the microscope as known to the early my-
cologists, and. know it also in the culture tube. It must be isolated as a
pure culture and its life history and reactions followed out upon laboratory
media. Its definite place in some one of the aggregate species or groups of
Aspergilli should be thoroughly established; then, by careful study and
comparison proper nomenclature should not prove difficult.

This manual, then, seeks to serve two purposes: (1) to provide the worker
encountering an Aspergillus with means for its identification, and hence to
open to him the whole literature of the group, as well as the particular
species; and (2) by enumerating all forms found in the literature, and
indicating their proper allocation, to guide the user of that literature in
the interpretation of names found in his reading but not known to him in
nature, in culture, or in exsiccati.

The authors acknowledge the cooperation of Dr. Johanna Westerdijk,
Dr. F. H. van Beyma, and their colleagues at the Centraalbureau voor
Schimmelcultures, at Baarn, Holland, in the free exchange of cultures and

PREFACE IX

information. Professor Ph. Biourge and Dr. Paul Simonart at Louvani
put their entire collection at our service after preparing and demonstrating
their interpretations in their own laboratory. Dr. Raoul Mosseray sent
his extensive series of variants in the Aspergillus niger group as accumulated
from the Belgian Congo. Dr. Adalbert Blochwitz made many comments
and criticisms in his numerous letters. Professor Harold Raistrick and
Mr. George Smith submitted all strains reaching their laboratory with
full notes on their own interpretations. Cultures and photographs, some
of which are used in this manual, have been furnished by Messrs. John and
Edward Yuill. Series of cultures have been received from Drs. Marie
B. Morrow and J. J. Taubenhaus in Texas, Drs. Roberta Ma and Y. K.
Shih in China, Drs. G. Kita, R. Xakazawa, J. Hanzawa, and K. Oshima in
Japan, Drs. G. R. Bisby and G. A. Ledingham in Canada, and Dr. H. Macy
in Minneapolis, as well as individual strains from many correspondents.
The laboratory collection owed much of its completeness to the punctilious
workmanship and painstaking scholarship of Dr. Margaret B. Church.

In the preparation of the manuscript outstanding contributions have
been made by Dorothy F. Alexander who prepared the line drawings and
assisted generously in the checking and proofreading of the textual material;
by Mr. Roland W. Haines, Photographer of the Northern Regional Re-
search Laboratory, who made all the color pictures, as well as many of the
black and white photographs; and by Miss Nancy Brant who typed the
manuscript in its final form.

The authors are indebted to the Chas. Pfizer and Company Inc., Brooklyn
New York, for underwriting the cost of reproducing the natural color
photographs.

Administratively, the preparation of this manual was made possible by
the vision of Dr. O. E. May, Chief of the Bureau of Agricultural and
Industrial Chemistry, Mr. H. T. Herrick, Director of the Northern Re-
gional Research Laboratory, and Dr. Robert D. Coghill, Chief of the
Fermentation Division of the Northern Regional Research Laboratory,
who have developed the industrial exploration of the biochemical utili-
zation of the fungi over many years.

The Authors

PART I

GENERAL DISCUSSION

Chapter I
HISTORICAL INTRODUCTION

Historically, the Aspergilli, as a part of moldiness of things, have always
been a factor in man's environment, but for ages were brushed away as
white, yellow, green, red, or black mold, with or without any attempt at
interpretation. After the development of the microscope, men began to
see structure. Micheli (1729) distinguished conidiophores and heads.
He noted that the heads were rough, the spore chains or columns pro-
ducing an uneven surface, hence he gave the name Aspergillus (rough
head). He then marked with Latin phrases his sketches of differently
colored moldy substances, for example, Aspergillus capitatus ochroleucus,
probably some strain of Aspergillus ochraceus; Aspergillus capilulo pulla
for a black form, etc. Other authors followed, using much the same ter-
minology, but without illustrations definite enough to give knowledge of
the structure of the heads. Thus, Haller, in 1742, put what appears to
have been Sporodinia into the genus as A. ramosissimus, etc. There is
just about enough certainty in the use of A. albus, A. niveus, A. capilulo
pulla, A . purpureus, etc., to justify the continued use of the name Aspergillus
after taking out of the aggregate the extraneous material thrown into it
by the very scanty microscopic examination given by the early mycologists.

Persoon (in the 1790's) threw the Aspergilli into his polyglot concept,
Monilia, based upon the production of spores in chains resembling strings
of beads. He made no record concerning their origin. Then Link in
1809 went back to Micheli and based his rejection of Monilia upon the
specification that these chains of spores must have their origin in a "head"
(capitulum)

Link failed to examine that head closely enough to keep out questionable
forms although we know that in describing A. glaucus he had under his
microscope one of that group as it was found, then and now, upon partly
dried herbarium specimens. Correct interpretation of the structure of
this head appears first in the work of Corda who began, about 1828, to
publish his studies of fresh material, as seen under his microscope. Up
to about 1850 each worker was prone to look at his predecessor's descrip-
tions and figures, and either assign whatever he had to another man's
species, or conclude that each specimen he had was new and add another
group of names. Montagne complained (1856) that none of the descrip-
tions written before Corda were identifiable, while some of us are equally
uncertain of our ability to interpretMontagne.

3

4 A MANUAL OF THE ASPERGILLI

DeBary's laboratory in the early 1850's seems to have introduced suffi-
cient culture of the molds found to form the beginning of a permanent
literature. This started with the recognition that the yellow perithecia,
called Eurotium herbariorum by Link, which developed among the heads
of Aspergillus glaucus upon his herbarium specimens, were actually borne
upon the same mycelium (fig. 7). Fresenius, Cramer, Wilhelm, and Brefeld
in Germany followed. Raulin and van Tieghem in France developed the
fermentation of the tannins in gall nuts to gallic acid in the 1860's with
comparative study of other molds as a corollary. In 1880, in Paris,
Bainier began publishing his studies of molds as they appeared in phar-
maceutical products. He was followed by Gueguen, the Sartorys, and others
in France, and somewhat later by Biourge in Louvain, Belgium.

Wehmer, in Hanover, began publishing his biochemical studies in 1891,
which led him to develop his more pretentious monograph published
in 1901. Blochwitz undertook to develop his "system" early in the new
century, but the World War delayed its publication until 1929. Meanwhile,
Thorn and Church, beginning about 1910, had published The Aspergilli
as a taxonomic monograph in 1926. Aspergilli were listed in cryptogamic
floras, lists, manuals, and special papers of many kinds over the whole period,
but critical discussions were few.

In somewhat over 200 years, an enormous mass of Aspergillus literature
has accumulated. Justice to the writers at each stage in the development
of our information calls for an analysis of the conditions which surrounded
its development. Practically all of the early literature was microscopical:
the worker confined his study to specimens brought in from natural sources,
each of which was often assumed to be typical of some species. Each
worker used the microscope that he had at hand and the technique of
study already known to him. Life histories and comparative examination
of material from many sources were disregarded. Publications appeared
as parts of floristic studies of particular regions, as reports of organisms
found in particular lesions of man or animals, or as observed in special
industrial connections. After DeBary's group began to study organisms
in comparative culture, the number of publications began to increase
rapidly. By 1929-1930 Tamiya and Morita were able to cite 2,424 titles of
papers which, in some way, concerned the Aspergilli, in their published
Bibliographie von Aspergillus, 1729 bis 1928. A mathematical analysis
of this literature was published by Tamiya in 1931. Referring to his
table 1, 71 titles appeared in the 125 years before DeBary's 1854 paper;
73 appeared in the next 18 years preceding Brefeld 's 1872 papers; 236 in
the next 19 years just preceding Wehmer's oxalic acid reports in 1891.
All of this may be called the period of physiological morphology. The
remaining two thousand, published between 1891 and 1928, represent the

HISTORICAL INTRODUCTION

pure culture period. This may equally well be called the biochemical

period.

The taxonomic part of this literature was scattered through several
languages and represented many schools of nomenclatorial thought. The
man who had seen only three or four Aspergilli found no difficulty in sep-
arating them. Each used his own descriptive terms— adequate for his
purpose but useless to the next man with different species. Saccardo
just published them all. Critical analyses were not available.

In The Aspergilli (1926) as a monograph, Thorn and Church sought to
bring together all of this taxonomic literature, as published before that
date, and to present a critical opinion as to the proper relationship of the
species described, whether retained in the genus or placed elsewhere.
Some 350 names were thus accounted for, but the actual number of species
accepted as known in culture or probably determinable from existing
literature was given as 69 (p. 252). These were more or less arbitrarily
considered in 11 groups. In undertaking to account for all the described
forms, it was deemed advisable to include, in the various groups discussed,
many forms whose published descriptions were inadequate for positive
identification, but complete enough to indicate their affinities with known
sections of the genus. Citation of these species in the older literature might,
therefore, be traced to group relationship, and in that way, correlated with
more recent studies of the same or related organisms. In addition, certain
names were listed as entirely unidentifiable and certain other forms as
belonging to other genera.

Various other proposals for this purpose have been made. Blochwitz
in 1929 published his long-delayed "System und Phylogenie," with inter-
pretations and proposals for grouping quite different from those of Thorn
and Church. Neill (1939) reduced the species recognized to the larger
aggregates, paying little attention to details of head and spore formation.
George Smith (1938), seeking industrial utility, simplified his descriptions
and introduced many photomicrographs. He discarded the literature
for the most part and undertook to guide the worker to the larger groups
which could be located principally by color and shape of head, as shown by
his figures. Dodge (1935) keyed all species whose names appear in medical
literature, from their descriptions but without studying them in culture.
In 1939, Biourge prepared a manuscript analysis of the genus for the
Third International Microbiological Congress in New York. His associate,
Dr. Simonart, came to represent him, but left because of the war. The
paper was not presented but was transmitted to us because return to the
author was impossible. Biourge died somewhat later. His scheme of
classification prepared in his last years is not presented because it contains
many things too bizarre to do justice to a man who for many years was a
master workman, as well as a valued friend.

Chapter II

CLASSIFICATION, GENERIC DIAGNOSIS, AND

SYNONYMY

Class: Ascomycetes

Order: Plectascineae

Family: Aspergillaceae
Genus: Aspergillus

Class: Fungi Imperfecti

Subclass: Hyphomycetes
Order: Mucedineae

Family: Mucedinaceae

Subfamily : Aspergilleae
Genus: Aspergillus

The above classification follows Engler and Prantl. Changes in the
names of class, order, and family appear in various proposals without
essential differences in placement. G. W. Martin would replace the names
Plectascineae with Eurotiales, Aspergillaceae with Eurotiaceae, and Asper-
gillus with Eurotium in the plea that the first name applied to the ascosporic
form determines the generic usage. Since the group has too many common
characters to be split to advantage, and since the non-ascosporic forms
vastly outnumber the ascosporic, it is better to forget Eurotium along
with the technicality. In this arrangement, the name Aspergillus appears
in its proper place among the ascosporic fungi. It also appears among the
Hyphomycetes properly keyed to facilitate the identification of organisms
obviously related but which do not produce ascospores as far as known.

GENERIC DIAGNOSIS

<

There is progressive need for broadening the application of the name
Aspergillus to include organisms whose structures, as determined in
culture by microscopic study, point to membership in specific natural
groups. There is need for analysis of the question whether the whole
group shall be retained as Aspergillus or further divided into more closely
related entities, such as Eurotium of Link, Aspergillopsis of Spegazzini,
Diplostephanusoi Langeron, Sterigmatocystis of Cramer, and perhaps others.
There are so many arguments for keeping them in a single group that the
characterization of the genus Aspergillus used by Thorn and Church in

6

CLASSIFICATION, GENERIC DIAGNOSIS, AND SYNONYMY 7

1926 has been emended and introduced here. This is followed by brief
considerations of the other more significant synonyms.

Aspergillus Micheli, in Nova Plantarum Genera, p. 212, Plate 91. 1729.

Compare Link, in Obs. p. 16. 1809; Corda, in Icones Fungorum

4:31, Tab. VII, fig. 94. 1840; and Thorn and Church, in

The Aspergilli, p. 4. 1926.

Vegetative mycelium consisting of septate branching hyphae, colorless,
bright colored, or in a few forms slowly becoming brown in localized
submerged areas, or producing brown crusts, or sclerotia; conidial apparatus
developed as conidiophores and heads from specialized, enlarged, thick-
walled hyphal cells (the foot-cells) producing conidiophores (stalks) as
branches approximately perpendicular to the long axis of the foot-cell
and usually to the surface of the substrata in or upon which they are borne ;
conidiophores unseptate or septate, usually enlarging upward and broaden-
ing into elliptical, hemispherical, or globose fertile vesicles bearing fertile
cells or sterigmata either parallel and clustered in terminal groups, or
radiating from the entire surface; sterigmata either in one series only, or
as a primary series, each bearing a cluster of two to several secondary
sterigmata at the apex; conidia varying greatly in color, size, shape, and
markings, successively cut off from the tips of the sterigmata by crosswalls
(not produced by budding), and forming unbranched chains arranged into
radiate (globose) heads or packed into columnar masses; perithecia found
in certain groups only, unknown in most species, cleistocarpic, thin-walled,
producing asci and ascospores within a few weeks; sclerotia regularly found
in some strains, occasionally found in other strains, and not found in other
and closely related strains, mostly globose or subglobose, composed of
polyhedral thick-walled cells.

Eurotium Link, in Obs. p. 31, Taf. 2, fig. 44. 1809.

Synonym : Mucor herbariorum Wiggers, in Primitiae Florae Holasticae
as No. 1158. 1780. See also DeBary, in Bot. Ztg. 12: 425.
1854.

The yellow perithecia suspended in networks of hyphae above or at the
surface of his badly dried herbarium specimens were taken by Wiggers
(1780) as the basis of Mucor herbariorum. Link (1809) recognized the
bodies as ascosporic, hence segregated them under the generic name
Eurotium. Then in 1854 DeBary published proof that these perithecia
were borne upon the same mycelium as the asexual A. glaucus fruits among
which they developed. He then called each of his Aspergilli, Eurotium
Aspergillus followed by the specific name, whether ascosporic strains were
known or not. In spite of technicalities invoked by some to bolster the

8 A MANUAL OF THE ASPERGILLI

use of the name Eurotium for all Aspergilli, or failing in that, for all ascosporic
strains, most workers have accepted the numerical predominance of the
non-ascosporic strains as ample reason for the general use of the name
Aspergillus. For practical purposes, Eurotium is not used here.

Sterigmatocystis Cramer, in Vrtljschr. Naturf. Gesell. Zurich Jahrg. 4,
Heft 4, p. 325, Taf. II, figs. 1-15. 1859. *

Cramer, in 1859, published his study of a black Aspergillus from the
human ear. Since the fruiting head differed from that of Fresenius'
A. fumigatus by showing a primary series of sterigmatic cells radiating
from the vesicle, each bearing a crown of several sterigmata, which in
turn each bore a chain of spores, he made this character the basis of his
new genus Sterigmatocystis. Cramer's name has been accepted by many
workers, but was rejected by Wehmer, Thorn, and others on the proof that
such use would separate strains obviously related in such a group as
Aspergillus flavus and its allies, and even among the black Aspergilli
studied by Cramer himself. The additional name serves no useful purpose
as an aid to identification; hence is not recognized here.

Euaspergillus Ludwig, in Lehrbuch des niederen Kryptogamen p. 258.

1892.

The proposal to apply a separate generic designation to all Aspergilli
producing sclerotia would take out the groups typified by A. candidus,
A. niger, A. wentii, A. tamarii, A. flavus, and A. ochraceus. No one has
followed Ludwig.

Aspergillopsis Spegazzini, in An. Mus. Nat. Buenos Aires Ser. 3, 13:

434. 1911.

The black-spored Aspergilli were described as dematiaceous, hence
separated from all the other groups. No practical reason for accepting
this proposal has been offered.

Diplostephanus Langeron, in Compt. Rend. Soc. Biol. Paris 87: 343-345.

1922.

Under this proposal ascosporic Aspergilli with the double series of
sterigmata would be separated with A. nididans Eidam as type. No
technical application of nomenclatorial rules justifies the complications
introduced.

In addition to the above names proposed to cover blocks of species
with particular characters in common, a series of names have been used

CLASSIFICATION, GENERIC DIAGNOSIS, AND SYNONYMY 9

by various authors for individual species: Alliospora for a black form;
Ascophora nigrans for A. niger; Aspergillopsis Sopp for an unidentified
organism; Cladosarum for Yuill's mutant of A. niger; Dimargaris for some
white forms; Emericella and Inzengaea for A. variecolor; Mucor as a place to
assign A. herbariorum; Sartorya for a possible ascosporic A. fumigatus.
These names are cited in the check list but contribute nothing to this
study of the group as a whole.

Chapter III
MORPHOLOGY AND DESCRIPTION

INTERPRETATION OF PUBLISHED DESCRIPTIONS

Basic Assumptions: In interpreting the descriptions of Aspergilli in a
literature covering a long period of time, certain assumptions, although
not always justified, form a working hypothesis for presumptive identi-
fication. Conidiophore walls and conidial walls are assumed to be color-
less and smooth, unless color or markings are either figured or described.
Perithecia and sclerotia are assumed to be lacking unless the presence of
such structures is specifically noted. Colors are assumed to apply to the
general color scheme of the colony, unless specifically applied to the
conidia, ascospores, or other details by the describer. Whereas colony
coloration may arise from an admixture of conidial structures and varying
amounts of vegetative hyphae, colored or uncolored, together with perithecia
or sclerotia in greater or lesser numbers, it is assumed to result from the
massing of conidial heads unless otherwise stated.

Difficulties encountered in interpreting descriptions based upon color
are less for the worker with a growing culture before him than for the one
handling descriptive literature alone, since the presence of white, green,
yellow-green, brown, or black heads is readily distinguished with a handlens,
even though sparingly produced upon a colony in which another color
predominates as in many members of the A. glaucus group, or in A.fiavipcs.
The color of the conidial heads is often made the primary basis of species
description.

Extent of Study: Interpretation of descriptive literature accompanied
and supplemented rather than preceded the study of great numbers of
cultures so that the groups established are based upon the actual handling
of thousands of cultures representing hundreds of forms of Aspergillus
handled during a period of more than thirty years. Many of these were
studied on natural substrata before their isolation. In addition, exami-
nation of exsiccati from several large herbaria, while more or less unsatisfac-
tory as to detail in identification of species, furnish confirmatory evidence
of the soundness of the groupings proposed.

Types: For a few of the specific names in use today, the type strain has
been definitely maintained in culture. For most series, selection of a
morphological entity to give a concrete concept back of the use of a name
becomes a matter of critical judgment. For the purposes of this manual,
an attempt has been made to base the use of the individual name upon the

10

MORPHOLOGY AND DESCRIPTION 11

morphological picture most frequently encountered, rather than upon a
selected strain assumed to be, but not known to be, the one first described.
Descriptive terms: For purposes of description, a standardized use of
terms has been adopted. Great diversity is encountered in the literature
in various languages. Even translated into Latin, Saccardo never homo-
genized the terms so that succeeding descriptions upon the same page use
descriptive terms in the same sense. To make comparison with existing
literature more convenient, the usages defined in the following pages
will cover the morphology as definitely as possible and indicate the usages
found in the older literature. To serve as a basis for the collection,
interpretation, and presentation of pertinent information regarding
Aspergilli to be studied, a guide sheet of the type used by the authors is
presented in the introductory portion of the manual proper (p. 82).

THE ASPERGILLUS COLONY

Since few of the Aspergilli regularly produce perithecia and ascospores,
a basis for identifying the majority of the molds of this group as they are
actually encountered in nature and in culture must be found in the de-
scription of the colonies and in the details of morphology found in the spore-
bearing structures available.

The vegetative mass of most Aspergilli consists of submerged mycelium
from which only fruiting hyphae rise above the surface. Such colonies
suggest a field of ripening grain, in which conidiophores and ripening heads
predominate, and have been described as velvety (the German term used
is rase) from their appearance in many species. Some species produce a
more or less aerial felt (floccosity) of branching and interlacing hyphae
bearing conidophores. This is characteristic of certain strains of the A.
versicolor and A. fumigatus groups upon Czapek's solution agar and other
culture media commonly employed, and it is normally one of the most
striking characters of A. wentii under laboratory cultivation (fig. 1 B).
Many strains of the A . glaucus group form long streamers of hyphae hanging
from meat stored in cool, damp rooms and certain of these retain this
character in laboratory culture. The character of the surface growth is
a diagnostic characteristic which is usually fairly reliable under reasonably
uniform conditions of culture.

In describing the Aspergillus colony, cognizance should be taken of such
factors as age, rate of growth, temperature of incubation, and the composi-
tion of the substratum. Provided with this information, subsequent
investigators can intelligently interpret their cultures in terms of species
previously described.

Many species and strains of Aspergillus often produce conspicuously
zonate colonies. Most commonly, these take the form of fairly regular

Fig. 1. Colony types in the Aspergilli. A, Aspergillus parasiticus NRRL No.
465, heavy sporing, non-floccose colony on Czapek's solution agar, room temperature,
10 days, X 2. B, Aspergillus wentii NRRL No. 375, light sporing, floccose colony
grown under the same conditions, X 2. C, Aspergillus variecolor NRRL No. 1954
characterized by the production of abundant, large perithecia, X 3. D, Aspergillus
alliaceus NRRL No. 315 characterized by abundant black sclerotia, X 2. E, Asper-
gillus ochraceus NRRL No. 408 on Czapek's solution agar, showing heavy sporing,
essentially azonate colony, X 2. F, The same on hay infusion agar, strongly zonate,
X2.

12

MORPHOLOGY AND DESCRIPTION 13

concentric zones and result from an increased production of conidial
structures periodically during the development of the colony (fig. 1 F).
The exact conditions and factors responsible for their appearance has not
been determined, but they obviously develop, in some way, as a response
of the fungus to its environment. Particular strains cannot generally be
described as either zonate or azonate, since the same organism exhibits
both characters at different times and under different conditions. In
contrast to concentric zones, certain species in laboratory culture char-
acteristically develop conidial heads in localized areas only. This is
particularly true of members of the A. glaucus group, and the A. sulphur eus
series, with conidial heads normally more concentrated at the margins of
slant tube cultures.

Coremia, in the broader sense of ropes of hyphae anastomosing and
trailing upon or near the surface of a substratum, occur more or less
commonly in Aspergilli of certain groups; but as specialized erect aggrega-
tions of conidiophores (StilbumAike) , such structures are described and
figured only for A. vitelline/, of Ridley. Since Ridley apparently de-
scribed his form only as collected upon a natural substratum, the actual
development of a specialized structure in this form remains doubtful.
Ridley's figure could be repeated many times by roughly drawing masses
of conidiophores bursting through the otherwise unbroken surface of a
rich nutrient substratum, such as seeds of cereals, producing a dense
cluster of conidiophores.

COLOR

The most striking character of an Aspergillus colony is usually its color
production. This takes two general forms: (1) color in the aerial parts,
including hyphae, conidiophores, heads, and conidia; (2) colors appearing
in the substratum, and representing the specific response and effect of the
organism upon particular media. The former is universally used in the
characterization of species, while the latter usually furnishes additional
pertinent data. In our study of the Aspergilli, citations of color have been
made according to the terminology employed by Ridgway since the
publication of his "Color Standard and Nomenclature" in 1912. This
terminology is used consistently in the present manual.

Group Color

There is a characteristic range of colors for each group (or collective
species) of Aspergilli, with a much narrower range in successive cultures
of the particular species or strains. The coloring substance may be de-
posited in the conidia only, as in A.flavus; in the conidial wall, and more or
less present in the sterigmata, vesicle, and upper part of the conidiophore

14 A MANUAL OF THE ASPERGILLI

as in A. niger; or the heads may be uncolored or nearly so, while the outer
layers of the conidiophore wall may be colored as in A. flavipes and A.
ochraceus. In some species of the A. glaucus group, the colony color is
at first green with the development of conidia, then predominantly yellow
to ferrugineous from ripening penthecia. Again, in other species of the
same group, the walls of the conidiophores and, more particularly, aerial
hyphae become encrusted with granules that are characteristically yellow
in the young colony, but become reddish or ferrugineous in age. Such
colonies are at first predominantly yellow with green heads inconspicuous
on a yellow background and latei become rusty red or brown.

In the A.flavus group, Saito (ly07) followed by Thorn and Church (1921,
p. 115) found that cultures with brighter shades of green when subjected
to a vapor of ammonia would lose the green color and assume the somber
yellow shades of variant members of the same group, and that this reaction
was reversible, since the vapor of acetic acid would restore or even intensify
an original green shade. The experiments pointed to the hypothesis that
the wide range of shades produced by mixtures of yellow and green in the
A. flavus-oryzae group' may be attributed to racial limitations, strain by
strain, in the range of hydrogen-ion concentration produced by metabolism.
When a carbohydrate fermentable by the particular species is present, an
acid reaction is promptly produced in the growing colony; as growth pro-
gresses alkaline products are also produced. The colony color in this series,
therefore, reflects first the intensity of the initial acidity as shown by the
intensity of the green color reached. The persistence of this shade or its
subsequent reduction or entire disappearance to leave a somber yellow or
finally brown colony, represents the balancing of the two activities. As
a result, certain strains of this group, if grown on Czapek's solution agar,
are quickly and very persistently deep green, others become particular
shades of green, which fade to yellow and finally some of them to brown,
and a few forms produce no true green color but assume a somber yellow
with the first development of conidia.

Color changes in the conidia have not been satisfactorily worked out.
In the A. niger group, the shade of yellow to purple-brown or black seems
to be a strain or race character little influenced by handling which is not
destructive of the racial entity. Each race seems to reach a fixed quanti-
tative limit in the secretion of the coloring substance, thus reducing the
most conspicuous diagnostic character among closely related forms to a
quantitative rather than a qualitative basis of separation.

Color in Conidial Walls

The conidial walls may be smooth but carry sufficient coloring matter
in diffused form to give the characteristic colony color. Within such

MORPHOLOGY AND DESCRIPTION 15

series as .4. fumigatus, A. nidulans, and A. ochraceus, however, strains or
races may be found which vary from conidial walls smooth or nearly so,
to walls bearing echinulations or even traceries apparently produced by
aggregation of color substance into spinules, or bars between the outer
and inner walls of the cells. Although smoothness and echinulation have
received much weight in descriptive literature, observations such as the
above would indicate that this character should only be used to separate
nearly related strains in the same group, rather than as a group character.
Literature on the nature of color in Aspergilli seems to begin with
Linossier's study of aspergilline as produced by A. niger in 1891; this was
followed more recently by Quilico, and Quilico and Di Capua in 1933.
Disregarding the record of observations only, more pretentious work
appeared when Bainier and Sartory undertook to use color production to
separate members of the A. glaucus group about 1910 to 1912. Their
experiments supplemented observations of colonies checked against a
color chart, with a routine series of solubility and precipitation tests.
Blochwitz (1929-1935) followed by using a series of routine solubility tests
against all species producing bright colors but failed to coordinate his
tests to show the relation of test to culture medium and conditions and to
age of the culture studied. Later Gould and Raistrick (1934) and Raistrick,
Robinson, and Todd (1937) studying the A. glaucus group, extracted and
defined the colors found, but again failed to follow the transformations
in the color of the particular species during the course of colony development.
Until someone correlates color determination and composition more
closely, color observations will continue to be useful accessory data which
must be related to the age of the colony and to the composition of the
medium as closely as possible to have value.

Colors in the Substratum

Production of bright colors in the substratum is frequent among the
Aspergilli. The color produced by any species or race in any medium is
dependent first on the ability of the mold to elaborate the particular
product, and second on the presence of the necessary building material
in the substratum. A mold may grow well upon a particular medium
without discoloring it; a transfer from this colony to another substratum
may turn the second medium red or yellow.

Color in the substratum is the result of the particular Aspergillus acting
upon the particular medium under a certain range of temperature. Most
of the species of Aspergillus, if they produce any color, produce from a
trace to abundant yellow in the early stages. This may persist, or give
place to shades of orange, red, or purple. In some cases, the color fades
out as the colony becomes older. In descriptive work, progressive changes

16 A MANUAL OF THE ASPERGILLI

in intensity of color, or the presence or absence of color in different sub-
strata, make the use of closely defined shades or intensities of color in the
substratum an unreliable means of characterizing cultures.

Observations of colors produced in the substratum remain, however,
very conspicuous and exceedingly useful accessory characters which aid in
the placing of species. The describer must bear in mind that such color
reactions are confirmative, not absolute characters in separating species.

The final difficulty in dealing with color as a separating character rests
in the loose use of color names, which is only partially corrected by the
use of color standards. Comparison of the same culture by different
individuals introduces very considerable discrepancies which become
serious when a specific descriptive name or number from one of these
standards is introduced into a technical description. In general, a series
of observations giving a range of colors for a species is less liable to introduce
errors of subsequent identification.

MORPHOLOGY

From the time of Micheli, the name Aspergillus (literally, rough head)
has been used for molds with a conidiophore or stalk and spore-bearing
head (capitulum).

The Head

The first structure observed in a detailed study of the colony is the
spore-bearing head. The color, shape, size, and arrangement of such
heads are characteristic of the species and to a lesser extent of the groups
to which they belong (figs. 4 and 5). Wehmer (1901) roughly grouped
his Aspergilli into Microaspergilli and Macroaspergilli on the basis of
the size of the fruiting parts. Thus, A. fumigatus, A. nidulans, A. sydowi,
and A. versicolor would represent Microaspergilli, while A. niger, A.
clavatus, A. ochraceus, A. wentii, and A. tamarii would be readily classed
as Macroaspergilli. The distinction breaks down when great numbers
of forms are studied, but the comparative size of heads and conidiophores
remains a useful adjunct in description. The heads in certain species
show a consistent range of measurements and form, as in A. fumigatus
and A. nidulans which have heads of small diameter forming columnar
masses (fig. 4). Similarly, characteristic heads of A. niger, A. ochraceus,
or A. wentii are globose and large (fig. 5). In other species, notably in A.
flavus or A. candidus (fig. 60), several sizes and shapes of heads are regularly
found in the same colony. The range of size and shape, however, remains
characteristic. The observation of many heads in the colony, and pref-
erably in many separate cultures, forms a better basis for description of
sizes and measurements than limited observation. Further description

Platk I

A-D, Aspergillus nidulans (Eidam) Wint., XRRL No. 193: .4 (upper left), portion of colony showing
developing peritheeia and abundant conidial heads; B (upper right), conidial heads showing typical color
and columnar form, X 60; C (center left), photomicrograph of conidial heads showing brown-walled conidio-
phores and green conidia, X 500; D (center right), ascospores, red in color and showing two equatorial ridges
when seen in profile, X 1200. E and F, Aspergillus janus Raper and Thorn, NRRL No. 1787: E (lower left),
single colony showing crowded short -stalked green heads in central area and long-stalked white heads in
localized marginal areas, the yellow-white, floccose areas being composed largely of hiille cells; F (Jower right >,
portion of same colony somewhat enlarged, X 8. (Color photographs by Haines, Northern Regional Research
Laboratory. Reproduced through co-operation of Chas. Pfizer & Co., Inc.)

MORPHOLOGY AND DESCRIPTION 17

of the structure of the head is given in the more logical order, following
the discussion of the stalk or conidiophore.

The Foot-Cell

The first step toward conidium formation in the Aspergilli is the differ-
entiation of certain cells (the foot-cells) in the mycelium for propagative
purposes. These cells become larger, thick-walled, and each usually bears
a single conidiophore as a branch, perpendicular to the long axis of the
cell (fig. 2 A) and usually about midway between the ends of the cell. In
age these cells frequently become fantastically curved and twisted with
their connection to vegetative hyphae inconspicuous but usually still
determinable. These foot-cells are commonly submerged in the sub-
stratum, although there are a number of strains of the A. glaucus, A.
fumigatus, A. versicolor, and especially of the A. flavus-oryzae groups in
which the conidiophores arise in this way from aerial hyphae. In A . effusus
the foot-cells are frequently long and several of them connected together
to form whole hyphae bearing considerable numbers of very short conidio-
phores (fig. 71 B3), hence their differentiation from the sterile or vegetative
cells is less easily determined. Failure to recognize the foot-cells as
present in the Aspergilli led Ferdinandsen and Winge (1920) to describe
S. dipus, using the foot-cell as the principal diagnostic character of the
species. The presence of such a differentiated foot-cell is proposed as an
arbitrary character to be used in separating certain depauperate forms of
Aspergilli, which approach the structure and appearance of the mono verticil-
late Penicillia (Citromyces), from the Penicillia. Organisms which lack
the typical Aspergillus head with its conidiophore and especially its foot-
cell may be best classified elsewhere.

The Conidiophore or Stalk

The erect, perpendicular branch from the foot-cell constituting the

conidiophore usually enlarges upwards toward the apex at which it dilates

more or less definitely to form the vesicle (fig. 2 C-E). The section of

the conidiophores from the foot-cell to the base of the conidial head is

measured and reported in describing species.

The conidiophore in some groups is not only septate, but each cell is
sufficiently distinct to justify the term articulate which is frequently
encountered in the older descriptions. In our experience in examining
specimens, articulate conidiophores are found only in the A. glaucus group.
In most Aspergilli the unity of the whole conidiophore is fairly accentuated.
Septa, if present, are thin, fragile, and inconspicuous; the whole conidiophore
is enclosed by continuous characteristically thickened walls without
conspicuous nodes as an evidence of septation.

18

A MANUAL OF THE ASPERGILLI

Thickening of the conidiophore wall may be uniform or may be greater
at the base, thinning to negligible toward the apex. Two general sections

Fig. 2. Development of the conidial apparatus in Aspergillus niger. A, Foot
cell bearing young conidiophore as a vertical branch. B, Developing conidiophore,
X 172. C and D, Development of the vesicle by swelling of the terminal portion of
the conidiophore, X 265. Ex and E*, Vesicle in optical section and surface view
showing early development of primary sterigmata, X 265. F , Later stage in develop-
ment of primary sterigmata, X 265. _G, Young fruiting head showing secondary
sterigmata bearing chains of conidia, X 265.

based upon the character of the thickening of the conidiophore wall are
fairly readily distinguished, although the careful use of high magnification
is occasionally necessary to separate certain strains. In the first, the

MORPHOLOGY AND DESCRIPTION

19

outer surface of the wall is free from pits, warts, or roughenings, hence is
called smooth; its structure is difficult to differentiate; the mass of the cell-
wall appears homogeneous or nearly so under the microscope and does
not absorb the ordinary protoplasmic stains. In some species the inside

*

■s, %. ^ W g§- — -Comd,a- ■

III gU#

Fig. 3. A, Aspergillus niveo-glaucus, NRRL No. 127, typical head showing only
one series of sterigmata, X 575. B,A. versicolor, NRRL No. 239, typical head show-
ing sterigmata in two series, X 1200.

surface of the wall shows irregular clumps or uneven thickenings. When
broken, many of these conidiophores show uneven or jagged ends like a
broken glass tube; in the A. niger group the broken ends split like bundles
of laths (fig. 64 B), giving a possible clue to the method of their formation.
In the second section, the wall appears dotted or pitted, or as interpreted

20

A MANUAL OF THE ASPERGILLI

Fig. 4. Conidial heads; group types. A, Aspergillus clavatus group: conidial
heads of A. giganteus, NRRL No. 10, showing typical clavate form, X 15. B, Asper-
gillus glaucus group: heads of A. niveo-glaucus, NRRL No. 127, showing character-
istic radiate pattern, X 35. C, Aspergillus nidulans group: heads of A. nidulans
showing typical short-columnar form, X 35 (Photograph by Edward Yuill). D,
Aspergillus flavipes group: heads of A. flavipes, NRRL No. 1959, typically barrel-
form, or loose columnar as shown, X 18. E, Aspergillus terreus group: A. terreus,
NRRL No. 265, heads columnar, of uniform diameter throughout, often becoming
quite long as shown, X 22. F, Aspergillus ustus group: A. ustus, NRRL No. 1974,
heads typically loose and radiate as shown, under certain conditions approaching
columnar, X 22.

Fig. 5. Conidial heads; group types. A, Aspergillus candidus group: A.
candidus, NRRL Xo. 308, showing characteristic mature heads of different dimen-
sions, X 18. B and C, Aspergillus niger group: B, strain NRRL No. 67, showing
typical mature heads, X 30; C, A. niger mut. schiemanni, heads typically globose as
shown (Photograph by Edward Yuill), X 30. D, Aspergillus wentii group: typical
globose heads of A. wentii, NRRL No. 397, X 18. E, Aspergillus flavus-onjzae group:
A. flavus, NRRL No. 1957, typical mature heads, X 18. F, Aspergillus ochraceus
group: A. ochraceus, NRRL No. 398, typical mature heads splitting into divergent
columns of conidia, X 18.

21

22 A MANUAL OF THE ASPERGILLI

with low magnification, often rough or echinulate (e.g., A. flavus-oryzae
group). The secondary thickenings in such conidiophores appear to have
been laid down around protoplasmic areas, which, for a time at least,
maintain contact with the primary wall outside. The size and abundance
of the pits in the mature conidiophore wall differ with the species, but in a
general way correspond with the rate of withdrawal of the protoplasmic
mass from its primitive connection with the original outer wall. In some of
the species in both groups, warts, or superficial and usually more or less
hemispherical concretions are found on the outer surfaces of the conidiophore
wall, sometimes few and scattered widely, again fairly numerous, but always
unevenly distributed (e.g., A. ochraceus group). These warts, or con-
cretions, appear to be deposits of excreted substance, possibly due to the
evaporation of the numerous drops or globules of liquid abundantly visible
upon the young and growing conidiophores.

The color of the conidiophore wall may be homogeneous, or the layers
may differ markedly in shade. In a number of the groups the entire
wall is hyaline. In certain other groups the outer layer is yellow as in
A. ochraceus and A. fiavipes, or it may be some shade of green, brown, or
avellaneous. In some species the whole wall is colored for all or part of
its length. No explanation of these color differences is available, except
possibly the varying concentration of aspergilline (Linossier, 1891) in the
upper part of the conidiophores of members of the A. niger group, as well as
in the conidial heads.

The Vesicle

The conidiophore is usually much larger toward the apex than at the
point of origin. At the base of the head a further dilation occurs more or
less abruptly to produce the vesicle (blase, of the German mycologists).
This vesicle is globose, hemispherical, elliptical, or long clavate in various
groups of the Aspergilli and furnishes an enlarged surface for the attachment
of spore-bearing cells. The lumen of the vesicle is continuous with that
of the upper part of the conidiophore; a septum near the base of the head
is occasionally, but only rarely, seen (see Corda, A. mucoroides for descrip-
tion), and has not been regularly found in any species.

Sterigmata (Compare Fig. 3)

The conidia-bearing surface, represented by the fertile area of the vesicle,
is closely covered by the simultaneous development of a layer of cells, the
sterigmata, each in a general way perpendicular to a point on the fertile
surface of the vesicle. In figure 3A a single layer of such cells is shown,
each of which produces an unbranched chain of conidia. In figure 3B
each of the first series of cells, or primary sterigmata, bears two to several

MORPHOLOGY AND DESCRIPTION 23

cells, the secondary sterigmata, forming a crown, or verticil, at the apex.
Each of the secondary sterigmata bears one chain of conidia. The mech-
anism shown in figure 3B would produce several times as many chains of
conidia as figure 3 A. Such a head as figure 3B would be compact, whereas
figure 3 A would represent a loose head.

In figure 3A the cells in the single layer, each producing a chain of spores,
are in the strict sense the sterigmata. In figure 3B cells of the first layer,
or primary sterigmata, produce verticils of cells, the secondary sterigmata,
each of which is in the strict sense a sterigma. The cells are usually char-
acteristic in size and shape for series of closely related species. Where
there are both primary and secondary series, the primary sterigmata are
essentially supporting cells and vary much more in size and shape than do
the secondary sterigmata. For this reason they are more useful in species
diagnosis than the secondary series.

Various usages are found in the literature. The primary sterigmata
are often called basidia. The secondary sterigmata are called phialids
because they have somewhat the shape of the pharmacists' phial (vial).
In translating descriptions into the Latin, Saccardo apparently followed
the describers verbatim, hence used no consistent terminology. We
find the primary sterigmata as sterigmata, basidia,. or pseudobasidia, and
the secondary series as sterigmata, pseudosterigmata, ramuli ("ramulis
sporiferis") or even rami (branches); all of these usages have been homo-
genized here into "primary and secondary sterigmata."

Conidium Formation

The actual spore-producing cell, or sterigma, is definitely specialized.
It ordinarily consists of an essentially cylindrical body, which, after
reaching a length more or less uniform for the species, narrows into a
spore-producing tube whose diameter is fairly uniform within the species.
Elongation is thenceforth confined to this spore-producing tube. The
nuclei in the sterigmata divide and one of each pair of daughter nuclei
passes into the tube; cell division follows. Parallel with the repeated
division of the sterigma nucleus, the tube continues to elongate rapidly,
successively cutting off new sections and pushing the older cells outward.
Each such chain of spores typically consists then of series of equal sections
cut from one tube or tip of a sterigma and each carries a daughter nucleus
derived directly from the active nucleus of the sterigmatic cell at the base
of the chain. No further divisions occur among the cells in the chains.
Such chains often contain several hundreds of spores, or conidia, each of
which is theoretically at least exactly like the rest, hence fully capable of
propagating the species (fig. 6).

24

A MANUAL OF THE ASPERGILLI

The Conidium, or Spore

The conidia are thus specialized propagative cells, asexual in origin,
produced by a complex cellular fruiting structure. This consists (1) of a
foot-cell connected with the vegetative mycelium and usually imbedded
in the moist substratum, (2) of a conidiophore, or stalk, rising more or less
vertically into the air to a distance typical of the species and enlarged at
the apex to form the vesicle which is the central unit, and (3) of a dilated
head consisting of one or two series of cells, the outermost of which are
specialized for the purpose of producing chains of cells (the conidia),
each equally capable of carrying the genetic factors necessary to propagate
the species.

Fig. 6. Camera lucida sketches showing progressive stages in conidium formation,
X 700. A, Initiation of conidium formation. B, Secondary sterigma bearing a
chain of three conidia, the outermost developing characteristic roughenings by the
deposition of coloring matter between the outer (thin) and inner (firm) wall. C,
Sterigma bearing a chain of conidia in which differentiation of the outermost spore
is complete. D, A single mature conidium seen in surface view. See discussion on
page 24.

After the conidium is separated by a septum from the mother cell or
sterigma, it remains so attached1 as to draw nutrients from the parent
cell at first, while it assumes the size and shape characteristic of the species,
then it lays down within its original, or primary wall, a secondary cell wall
whose color, texture, and marking are those of the species. The secondary
wall completely separates this spore from the parent cell. (fig. 6). Exact
uniformity is not attained. An occasional cell fails to develop; some differ-
ences in size are usually evident; markings, while characteristic in nature
and general pattern, are not always identical as to details. For descriptive
purposes, size ranges are therefore more important than exact measurements
and the nature of the markings found are more important than the relative

1 Buller (Researches on Fungi V, Chapter II, 1933) discusses the primary septum
as having a central pore through which connections are maintained.

MORPHOLOGY AND DESCRIPTION 25

number and dimensions of such. Such conidia may be very thin-walled,
delicate and readily destroyed, or firm-walled, almost impervious to stains
and able to retain their vitality for many years. They are extremely small,
light, and float readily in air currents. In many species, the outer layer of
the spore wall absorbs water slowly, hence such spores tend to float in
currents of fluid or to develop as mycelia covering the surfaces of liquid
media. Molds being typically aerobic, normal colonies develop only on
the surface of the substratum where oxygen is abundant and their spores
can be discharged directly into the air. Spores developing under submerged
conditions in the absence of adequate oxygen produce fragmentary and
defective mycelia only.

The "Connective"

Descriptive literature often cites the presence of a "connective" or
"disjunctor", a "bridge" between conidia in the chain. This is sometimes
present, again absent, in the same microscopic preparation, and when
seen, it appears as a short space between spores, bridged by transparent
cell walls. This is exactly what it is. Cells cut off from a cylindrical
tube may swell and assume subglobose form without breaking their area
of contact, or, in the swelling and rounding up process, they may partially
or completely break that contact leaving the original cell wall of the tube,
within which they developed, as a bridge across the open space (fig. 64 C).
The critical examination of the developing cells in thousands of preparations
have failed to justify interpretations which assume the degeneration of
every alternate cell, or fantastic fusions in the production of conidia. The
observation of connectives is, therefore, ordinarily worthless because
morphologically it means nothing, and it is not justified by successive
studies of the same species.

Endogenous Conidia

A spore is described as endogenous if it is formed within a tube or cell
wall of a previously existing cell. It may be extruded through a tube.
That tube may be used once only or many times. The critical factor
is the formation of a cell or spore within an existing specialized spore-
bearing organ and its extrusion from that body through a fixed tube. In
Aspergillus, the tip or tube of the sterigma elongates, a cylindrical section
is cut off carrying the tube wall and the septum at each end as the primary
wall of the spore itself. Within that primary wall the spore as an entity
rounds itself up to characteristic form, deposits or lays down its own wall
with whatever coloration or markings may be typical of the species. The
primary wall may remain separate and distinct and in the ripe spore be
visible under the microscope, it may be blended with the secondary wall,

26

A MANUAL OF THE ASPERGILLI

or it may not be determinable on the ripe conidium by ordinary examination.
In the sense of the definition above, no endogenous conidia appear in
Aspergillus. In cases, the primary walls are seen to break away if ripe
spores are mounted in fluid, often carrying with them the granular materials
which impart the characteristic marking to the spore, hence leaving the
wall of an .A. nigcr spore smooth (A. luteo-niger Lutz).

.«**.

•>

\

«

«L

--

)

•

£$&

Fig. 7. The development and relationship of Aspergillus glaucus and Eurotium,

after DeBary, 1854.

Perilhecia

The general morphology of the perithecium of the A. glaucus group
(Eurotium) was described and figured by DeBary (1854, 1870), while
that of A. nidulans was described by Eidam soon thereafter (1883).
DeBary described the yellow to orange or ferrugineous perithecia as from
90 to 300 m in diameter, without ostiole and without specialized appendages
(fig. 7). These perithecia are borne, above the surface of the substratum

MORPHOLOGY AND DESCRIPTION 27

and are never more than loosely hung in networks of hyphae; they are
often very abundant and dominate the color of the colony. Perithecium
formation is favored by abundance of assimilable carbohydrate, but may
or may not be completely suppressed by its replacement with nitrogenous
products ; transfers from the same culture have developed colonies yellow
from perithecia when grown on sugar solution, or dense green without
sign of perithecia upon a peptone medium or a piece of leather.

The perithecium of Aspergillus arises from a branch which coils in various
manners in the different species to become the ascogone, as figured by
Dangeard from forms reported as E. herbariorum, A. flavus, A. fumigatus,
S. ochracea, S. nidulans, by Fraser and Chambers for A. herbariorum, and
by Dale (1909) for A. repens. No fertilization process was demonstrated
in those studies. Dangeard (1907) reported the cells in the fruiting
apparatus as multinucleate in E. herbariorum, but to be mononucleate
in the other species figured. In general, however, the development of an
ascogone, followed by the development of the perithecial wall and accessory
cell masses from vegetative hyphae below the ascogone, has been roughly
described and figured for the group represented by A . glaucus and A . nidulans
but interpretation of the other specific names used by Dangeard is doubtful,
even for grouping, on the basis of the descriptions and figures given.

Henrard (1934), working with 15 "species" of Aspergilli, reports that
all of them are "sexually homothallic." He summarized the literature
by saying that homothallism in the Aspergilli has been affirmed by Kniep
(1928) for A. repens; by Schwartz (1928) for A. ruber, A. repens, four
other strains of "A. glaucus", and four strains of A. nidulans; by Blochwitz
(1932) for six strains in the A. glaucus group; and by Greene (1933) for
A. fischeri.

So far then, as present information goes, heterothallism cannot be assumed
to account for the great variability of the Aspergilli.

Perithecia are regularly found in most of the species of the A. glaucus
group and in A. fischeri, which is closely related to A. fumigatus, and in a
series of closely related forms in the A. nidulans group. They are not
sporadic or dependent upon unknown conditions, but are regularly pro-
duced in media which are adequately supplied with sugars and the salts
in routine use. Their presence in A. wentii and A. citrisporus was asserted
by Thaxter (personal communication) without producing either material,
or description of the ascospores. Dangeard (1907) claimed to have found
ascospores within the sclerotia of A. niger but failed to describe them.
Diligent search over many years has failed thus far to confirm either
statement. Still, it may be assumed that such sclerotia may be the
homologue of structures which under some conditions might become peri-
thecia.

28 A MANUAL OF THE ASPERGILLI

The Ascospore

So far as fully described, the ascospore of Aspergillus follows the general
type shown in the figures and description of DeBary. In the course of
its development, the secondary thickening of the cell-wall develops in
the form of two symmetrical valves suggesting the arrangement found
in the shell of a bivalve mollusk, such as the hard clam (Venus mercenaria).
The ripe ascospore is commonly shaped as a double convex lens with the
valves more or less closely in contact at the edges. A series of variations
upon this basic pattern occur and characterize particular species (figs. 27,
34, and 43). If the exospore is smooth and the margin of the valves is
not marked by folds or ridges, figure 27A characteristic of A. re-pens appears;
if the exospore is rough in the absence of marginal folds, figure 27D
characteristic of A . amstelodami results ; if the exospore is rough and the
margin of the valves bear folds or ridges, figure 43C characteristic of A.
rugulosus and A. jischeri is produced. An extreme development of the
marginal folds is seen in- A. variecolor, figure 43D.

When such an ascospore germinates (fig. 8A), the figure first shown by
DeBary (1854) develops. Exactly the same type of germination is shown
by A. nidulans, in which as the spore swells, the valves first separate at
one edge, then parting completely, remain on opposite sides of the germi-
nated spore conspicuously identifiable by the bright purple-red color of
the valves (Thorn and Church, 1926).

Htille Cells

Mature perithecia were described for A. nidulans by Eidam (1883)
but without giving attention to their origin. In his description, Eidam
pictured a loose network of hyphae more or less completely surrounding
the perithecium containing large numbers of "Hiille" cells, terminal or
intercalary cells which swell and become vesiculose, elliptical or almost
globose, then develop very heavy thick walls almost obliterating the cell
lumen (fig. 49). These cells were later noted by Dangeard (1907) who
designated them as chlamydospores, but failed to present evidence of
function as propagative cells. These cells are abundant in connection
with perithecia in all strains of the A. nidulans group, but are lacking in
A. unguis which does not produce perithecia. Cells of the same general
character appear in sterile masses of hypae in strains of the A. ustus and
A. flavipes groups (fig. 49), and in some strains of the A. versicolor group,
while thick- walled, septate hyphae at least suggestive of hiille cells appear
in Aspergillus carneus in the A. terreus group (fig. 49F).

In the .4 . nidulans group hiille cells, "Eidamsche blasen," or chlamydospore
(terms found in the literature) are always produced in connection with
perithecium formation (fig. 42). In the A. flavipes, A. ustus, and A.

MORPHOLOGY AND DESCRIPTION*

29

h

A

^f

9

i

^

•

a

b

©

^

rf

B

Fig 8 Spore germination. A, Germinated ascospores of Aspergillus nidulans
X 750: a, ascospore in profile showing how the two valves comprising the spore wall
are pushed apart as the spore content enlarges and the sporelmg develops; b, the
same in face view. B, Germinating conidia of Aspergillus niger, X 600: a, ungermi-
nated spore ; b , spore in early st age of germination ; c , germinated spore showing rapidly
elongating sporeling.

30 A MANUAL OF THE ASPERGILLI

versicolor groups, they occur in scattered aggregates of varying conspic-
uousness, although no perithecia have been found. Occasionally, as in one
of the A. flavipes group, foot-cells and sterile cells, which appear to be
conidiophores ending in points instead of heads, appear. These aborted
conidiophores vary from nearly full length to vestigial. From the study
of such material it is believed that Eidam's hiille cells represent aborted
conidiophores surrounding the perithecia of A. nidulans and they may be
indicative at least of a vestigial precursor of perithecium formation when
they occur elsewhere.

Hiille cells do not, however, accompany the perithecia of either the A .
glaucus group or A. fischeri. In the former, the perithecia are smooth-
walled and naked ; in the latter, they are surrounded by a loose network of
sterile but unspecialized hyphae.

In A. panamensis (Raper and Thorn, 1944) aborted conidiophores
commonly occur, but no hiille cells have yet been observed. In A. unguis
(Thorn and Raper, 1939), which is one of the A. nidulans group lacking
only the ascosporic phase, the long, sterile, thick-walled "spears" suggest
homologous structures. To summarize, hiille cells are specialized structures
that normally occur in certain groups of the Aspergilli. They are of
somewhat questionable origin and, in our experience, are not known to
serve any functional purpose. Schwartz (1928) figures the hiille cells of
A. nidulans as capable of germinating and thus acting as reproductive
cells. Since the cells of different groups are fairly characteristic, they
often provide valuable diagnostic group and species characters.

Sclerotia

Definitely hard masses with characteristic surface marking and color-
ation, and consisting of thick-walled parenchyma-like cells occur in several
groups of Aspergilli. Such structures have not been seen in the groups
typified by A. clavatus, A. glaucus, A. fumigatus, A. nidulans, A. ustus,
A. versicolor, or A. terreus. On the other hand, they develop regularly in
certain members of the A. candidus, A. niger, A. wentii, A. tamarii, A.
flavus-oryzae, and in the A. orchaceus groups; in other members of these
same groups grown under similar conditions, no such structures have
been found. In occasional strains of A. flavipes dark hyphal masses are
seen, but these are not sufficiently compact to be considered true sclerotia.
The development of sclerotia has not been followed in sufficient detail in
any species to fix their genetic significance, but the specialized, character-
istic structure of the sclerotia of many species lends color to the report of
ascospore formation by Dangeard (1907), although no one else seems to
have been able to verify such development. Environmental factors are
known to influence sclerotium development, but these have not been
carefully analyzed.

Chapter IV
CULTIVATION AND EXAMINATION

An Aspergillus occurring upon a natural substratum may frequently be
identified as to group from the original material if it belongs to one of the
large and well-marked groups. Among members of the same group, how-
ever, differences in the nature and composition of the substratum produce
marked contrasts in colony appearance, quantity of growth, coloration,
measurements of fruiting structures, and in the appearance of conidiophores
and conidial masses. Many Aspergilli can be identified as to group from
dried herbarium materials, but the process of drying generally changes the
colors materially and renders the hyphal masses so fragile that the morpho-
logical details necessary for identification inside a particular group are often
obliterated or made difficult to interpret. Although accurate placing of
such materials is sometimes possible, a much more satisfactory identifica-
tion may be reached by transferring fresh material to culture media of
known composition, followed by purification of the cultures so that they
present single species or strains for intensive study.

Two types of material for identification are regularly encountered by one
undertaking a study of the Aspergilli: (1) the culture already isolated by
another worker, and (2) the moldy substratum with its natural flora of
micro-organisms often representing several or many species, including
miscellaneous molds, bacteria, actinomycetes, and even protozoa and other
forms. In either case, the final decision as to the proper name of an
Aspergillus must usually be sought in fresh cultures made by the one re-
sponsible for identification.

Classification within the genus has become so dependent upon observa-
tion in pure culture that the whole subject of laboratory cultivation, includ-
ing favorable substrata, culture making, and culture handling needs to be
considered.

CULTURE MEDIA

Pure culture upon known substrata is almost essential to the identification
of Aspergilli. Since the morphological responses to diverse nutrients, and
especially to the stimulus of mixtures of other molds and bacteria growing
with any particular species, is great, the study of each strain or species in
pure culture in media of known composition is practically necessary.

Aside from Raulin's group in Paris, most of the early culture work with
Aspergilli was carried through upon so-called natural media. DeBary and

31

32 A MANUAL OF THE ASPERGILLI

Brefeld used decoctions of horse dung variously diluted and stiffened with
gelatin. Much of the European work was done with brewery wort; Maze
(personal communication, 1904) used extract of white beans; potato and
carrot decoctions have been widely used; Bainier grew his molds and de-
scribed many of them upon sticks of licorice root; others preferred plugs of
potato or of carrot, string beans, etc. The primary aim was to obtain an
optimum growth of the mold under observation, rather than to analyze its
relation to the substratum or furnish comparative data to distinguish it
from other members of a series.

An optimum culture substratum for comparative study of the Aspergilli
needs to contain the necessary chemical elements in the form of pure but
assimilable salts, supplemented by carbohydrates of such structure as to be
available to the largest number of species, and purchasable in pure form in
the chemical trade. Supplementary and often very important information
must be sought from variations in the proportions of nutrients used, or in
the introduction of widely different substances in replacement of particular
components of media already used. The Aspergilli as they are isolated from
nature are not very dependent upon vitamins or growth-promoting sub-
stances. For cultivation upon solid substrata, agar is almost universally
employed as a gelling agent.

When the study of a large number of molds is undertaken, comparison of
these molds under controlled and reproducible conditions of growth become
essential. Foremost among the conditions which must be standardized is
the culture medium, or substratum. Various authors have proposed
standardized and reproducible formulae which in their experience have pro-
vided uniform cultures over long periods, hence are of value for comparative
studies. A series of such media are presented.

Czapek's Solution Agar

As a routine medium for comparative work, the following formula origi-
nally adapted from Czapek (1902, 1903) by Dox (1910) has been widely
used. Minor variations in quantities apparently do not affect the reactions.
Cultural information given in this manual is obtained from growth upon media
produced by this formula unless otherwise specified.

Czapek's solution ar;ar:

Water 1 , 000 cc .

NaN03 3.0 grams

K2HP04 1.0 gram

MgS04-7H20 0.5 gram

KC1 0.5 gram

FeS(V7H20 0.01 gram

Sucrose (Cube or other good commercial grade) 30.0 grams

Agar 15.0 grams

CULTIVATION AND EXAMINATION . 33

To reduce caramelization the sugar is added just prior to final steri-
lization.

Czapek's solution agar is not offered as an optimum substratum for any-
particular species, but as a mixture approximately neutral in reaction,
which is readily made in any laboratory in fairly uniform manner, and which
permits moderately vigorous growth of nearly all of the saprophytic Asper-
gilli. The quantities of mycelium and conidia produced by many forms
are much greater upon other media, but for comparative study, a moderate
growth of the majority of the species is more useful than the great mass of
mycelium ard conidia which are readily obtained by using enriched sub-
strata.

Since the purpose of the Czapek formula is to insure the presence of the
chemical elements required in quantities sufficient to support good growth,
it is frequently modified as to quantities and nutrients introduced. For
some Aspergilli such as the A. glaucus group, the addition of 20 percent or
even 40 percent of sucrose has proved useful. Ammonium nitrate is some-
times substituted for sodium nitrate but with the loss of information as to
whether the mold utilizes the ammonia or the nitrate, or both. Dextrose
is commonly substituted for sucrose. The monobasic potassium phosphate
prevents precipitation of certain components in sterilization, but it produces
an acid instead of a neutral medium, hence complicates many pieces of work.
The introduction of peptone or yeast extract increases the sporulation of
some forms. These and other changes are made, however, by investigators
who still refer to Czapek's solution as the basis of their work.

Biourge, in his monograph of Penicillium (1923) and in his unpublished
Manuscript of Aspergillus (1939), put much emphasis upon the method of
preparing the neutral Raulin medium which he used for the growth of the
colonies analyzed in making his species diagnoses.

Neutral Raulin's Solution — Dierckx-Biourge

1. Magnesium carbonate 0.40 gram

Tartaric acid 0.71 gram

Triturate in a mortar with a few drops of distilled water and add quickly to a
flask of distilled water; make up to 100 ml.

2. To a liter flask with 800 to 900 ml. distilled water add:

Sucrose 46.60 grams

Ammonium nitrate 2.66 grams

Ammonium phosphate 0.40 gram

Potassium carbonate 0.40 gram

Ammonium sulphate 0.16 gram

Zinc sulphate 0.04 gram

Iron sulphate 0.04 gram

3. Add 66 to 67 ml. of the magnesium tartrate solution (1) to the mineral salt-
sucrose solution (2) and make up to 1,000 ml. with distilled water.

34 A MANUAL OF THE ASPERGILLI

In his detailed study of the A. niger group, Biourge's pupil, Mosseray
(1934a), gives his simplified Raulin's solution as follows:

Water, distilled 1,000 ml.

Sucrose 50 grams

Tartaric acid1 0.40 gram

Magnesium carbonate1 0.250 gram

Ammonium nitrate2 0.250 gram

Potassium carbonate2 0.40 gram

Ammonium phosphate (NH4)2HP041 0.40 gram

Ammonium sulphate1 0.20 gram

Iron sulphate1 (cryst.) 0.05 gram

Zinc sulphate1 (cryst.) 0.05 gram

Agar-agar1 20.00 grams

Sterilize 120° C. for 20 minutes.

Steinberg's Solution

Steinberg in the course of many years' investigation of a single strain of
Aspergillus niger (NRRL No. 334: Thorn No. 4247) developed a basic for-
mula for testing other phases of the nutrition of his mold. It is called the
"dibasic optimum" solution and carries mannitol instead of sucrose, sodium
nitrite instead of ammonium nitrate, with the addition of sufficient sulphu-
ric acid to obtain any desired reaction. Interpolations into this solution
offer many possibilities.

Steinberg's "dibasic optimum" (Thorn and Steinberg, 1939)

Water (distilled in Pyrex still) 1,000.0 grams

d-Mannitol 50.0 grams

i^aNO-: 2.0 grams

K2HP04 0.35 gram

MgS04-7H20 0.25 gram

FeS04-7H20 0.001 gram

ZnS04-7H20 0.00088 gram

CuS04-5H20 0.00020 gram

MnS04-4H20 0.00012 gram

NaMo04-2H20 0.00005 gram

H2S04 to pH 4.

A number of so-called natural substrata, including malt extract and hay
infusion agars, are very useful in the study of the Aspergilli. A great ma-
jority of species sporulate more freely upon malt extract than upon Czapek's
solution agar (fig. 9), and for this reason it is very useful where large quan-
tities of spores are desired. This medium is, however, less diagnostic than
Czapek's solution.

1 Merck reagents.

2 Kahlbaum reagents.

CULTIVATION AND EXAMINATION 35

Malt Extract Agar (Blakeslee's Formula, 1915)

Distilled water 1 ,000 cc.

Malt extract 20 grams

Peptone 1 gram

Dextrose 20 grams

Agar 20 grams

Add dextrose just prior to final sterilization. Conidial structures are
generally more numerous and are often borne on shorter conidiophores, and
there is an almost complete absence of coloration in the substratum. Ex-
cept in a few isolated cases, coloration of the spore heads themselves is not
materially altered. While the production of exudate in the form of drops
is not characteristic of many of the Aspergilli, it can be generally said that
droplet formation on malt agar is much less than upon Czapek's solution
agar.

Hay infusion agar is very useful in the isolation of Aspergilli from nature.
A 1 : 10 suspension of soil in sterile water is streaked on hay infusion agar
plates and incubated for one week to 10 days. Isolations are then made
from individual fruiting structures with the aid of a low-power binocular.

Hay Infusion Agar

Distilled water 1,000 cc.

Decomposing hay 50 grams

Autoclave for 30 minutes at 15 pounds. Filter.

Infusion nitrate 1 , 000 cc .

K2HP04 2 grams

Agar 15 grams

Adjust pH to 6.2±

No Aspergillus makes a luxuriant growth upon this medium, but a great
variety of forms make a limited development. Furthermore, such fruiting
structures as are produced are generally characteristic of the different
species present. It thus constitutes a very favorable substratum with
which to analyze and isolate the Aspergilli occurring in soils or other natural
substrates. The medium is likewise useful for securing limited sporulation
of certain forms, such as Aspergillus sparsus, which fruit very sparsely upon
Czapek's solution agar.

Czapek's solution agar enriched with peptone or corn steeping liquor is
often very useful. For example, in the cultivation of members of the
glaucus group, the addition of a limited amount of peptone greatly increases
the production of conidia, while addition of a small amount of corn steeping
liquor (e.g., 0.2 percent) increases the growth of most forms without mark-
edly affecting the character of the resulting colonies.

Fig. 9. Influence of substratum. A-C, Aspergillus variecolor, NRRL No. 212,
growing upon Czapek's solution, malt extract, and hay infusion agars, respectively;
10 days; room temperature. Note particularly the heavy development of perithecia
upon malt and the very sparse development of the same upon hay infusion agar.
D-F , Aspergillus caespitosus, NRRL No. 1929, growing upon the same media under
similar conditions. Upon Czapek's solution agar (D) dark hiille cell masses develop
along with a limited development of conidial structures; upon malt (E) and hay
infusion (F) agars hiille cell masses are lacking and conidial structures are very
abundant.

36

CULTIVATION AND EXAMINATION 37

SPORULATION MEDIA

In addition to the ordinary handling of molds in the laboratory, explora-
tion of their biochemical potentialities often necessitates the production of
considerable quantities of spores. Whereas some of the media listed above
may be employed for this purpose, it is generally advisable to employ
so-called "sporulation media." Requirements of different species and
strains vary and one frequently has to develop special solutions to meet the
needs of particular organisms under study. Such media may be used either
as liquid substrata or as solutions solidified with agar. In either case the
objective is the same: to secure the maximum production of spores with the
production of as little vegetative mycelium as possible.3

A number of sporulation media for use with different molds have been
developed by Dr. A. J. Mover. Three of these will be cited, while addi-
tional formulae may be found in the papers published by members of the
Fermentation Division, Northern Regional Research Laboratory.

Sporulation medium for A. niger {Moyer, Wells, Stubbs,
Hcrrick, and May, 1937)

Glucose 91 .3 grams

NH4N03 0.450 gram

KH2P04 0.072 gram

MgS04-7H20 0.060 gram

Beer 60.0 ml.

Distilled water to make 1 liter

This solution can also be used as a solid medium by the addition of 0.5
gm. per liter CaC03 and 30.0 gm. per liter agar. The above solution was
subsequently modified as follows (Gastrock, Porges, Wells, and Moyer,
1938):

Glucose 50.0 grams

(NH4)2HP04 0.560 gram

KH,P04 0.144 gram

MgS04-7H,0 0.120 gram

Peptone 0.20 gram

Beer 45.0 ml.

Distilled water to make 1 liter

3 For the surface inoculation of nutrient solutions in small flasks or other con-
tainers, dry spores in quantity can be removed from agar surfaces and floated on the
liquid surface by means of a 5 mm. loop. To inoculate from a liquid culture, tae
usual procedure is to remove a small portion of the heavily sporing mat, transfer
this to the solution, and dislodge the spores by vigorous agitation. Spores from
either type of culture can be suspended in water and used as inoculum in submerged,
or shaken, cultures. The addition of sodium lauryl sulfonate to the suspending water
in a concentration of 1 : 10,000 aids greatly in securing uniform spore suspensions.

38 A MANUAL OF THE ASPERGILLI

Sporulation medium for A.flavus (Moyer, Personal communication)

Glucose (commercial) 165.0 grams

Bacto-Peptone 1 .0 gram

MgSCV7H20 0.050 gram

KH2P04 0.060 gram

KNOj 0.500 gram

Fe (as tartrate) 0.040 gram

Agar 0.25 gram

Distilled water to make 1 liter

The above solution can be used as a solid medium by increasing the agar
concentration to 30.0 gm. per liter. (The small amount of agar included in
the basic formula is added solely for the purpose of increasing viscosity.)
Incubation should be at 22° to 24° C. This factor is critical for maximum
spore production with the kojic acid producing strain, NRRL No. 484
(Thorn No. 3538), for which the medium was developed. This medium can
also be successfully employed for spore production in A . niger by incubating
cultures at 30° C.

Abundant sporulation of many strains and species can be secured by cul-
tivation upon bread, whole cereal grains, or various types of milled products
of the same. Bread, if used, should not contain proprionates or other mold
inhibitors. The material to be inoculated should be moist but in no sense
wet, and special precautions must be taken to insure that the grain or bread
is properly sterilized before being used. The use of grain as a substratum
for molds dates back to prehistoric time in the fermentation industries of
the Orient where rice was, and still is, commonly used to produce the
"koji," or inoculum, used in the alcoholic and soya fermentation industries
of that area. In its classic usage, the molds cultivated were mostly mem-
bers of the A. flavus-oryzae group, but experience has shown that this
general type of medium can be used to advantage to secure heavy sporula-
tion of many other forms. From such material, series of surface fermenta-
tion flasks or other vessels can be uniformly inoculated by various means
involving aspiration of spores, or the direct transfer of heavily spore-laden
particles. Spore suspensions can be prepared and used for seeding sub-
merged cultures

TYPES OF CULTURE

Cultures for grouping and identification of the Aspergilli should be
grown in petri dishes. At the same time an adequate number of slanted
tubes should be inoculated and held in reserve as an uncontaminated stock
culture. Colonies so situated in the petri dish that they can be viewed
directly under low magnifications with the compound microscope are neces-
sary to supply a clear picture of the structure and course of development
of mycelium and fruiting parts. Such colonies can be obtained by several

CULTIVATION AND EXAMINATION 39

procedures, some of which have been developed for special tests, or for indi-
vidual mold problems. Practices useful in bacteriology are often applicable
to the problems of the mycologist. Selection of a procedure which satis-
factorily provides the information necessary to describe an Aspergillus calls
for discussion of a series of procedures commonly in use. Only in that way
may we show why some of these are adapted to the problems of identifying
an Aspergillus while others fail, for specified reasons, to furnish important
observations.

Spot Inoculations

Over long periods and in the hands of many investigators some type of
mass conidial inoculum has given dependable and reproducible results. The
most common method of transfer and, on the whole, probably the most
satisfactory one for maintaining a strain of Aspergillus, as well as other
molds, is the removal from the stock culture of a variable mass of conidia,
fruiting structures, or vegetative hyphae with some sort of needle or loop
and the transfer of this material to selected positions on fresh medium.
Practices differ. With the Aspergilli, colonies so placed as to permit radiate
development from the point of inoculation are most satisfactory for study.
Where there are two or three colonies to the plate, these eventually reach
into each other's zones of influence. They may blend and become indis-
tinct, or inhibit each other and leave sterile bands between the colonies.
Both types of culture furnish useful data. Usually, the line where two
colonies approach and partially or completely inhibit each other hastens
fruiting in the adjacent margins and permits favorable examination with the
compound microscope. The opposite margin of the same colony, unaf-
fected by competition, furnishes at the same time the normal and symmetri-
cal growth which is typical of the species. The one-colony plate usually
provides the most striking exhibit of the species, but the 2- or 3-colony plate
is the most generally useful (fig. 10).

Once an Aspergillus has been obtained in pure culture, the most effective
way to insure that plates will contain 1, 2, or 3 colonies, as desired, is to
suspend a quantity of spores in melted agar at approximately 45° C, allow
this to solidify, and then transfer small quantities of the gelled suspension
to the surface of plates or agar tubes in the positions where the colonies are
desired. With the Aspergilli, as with all of the molds which produce dry
spores, it is often very difficult to secure placement of colonies at selected
positions, and only at such positions, without the use of some type of
wetted inoculum.

In the routine examination of Aspergilli, where it is not essential that the
number of colonies be limited to 3 or less, satisfactory transfers have been
made by using a sharp nichrome wire as an inoculating needle and selecting

40 A MANUAL OF THE ASPERGILLI

the inoculum from a colony in a petri dish by working under a 10 X pocket
magnifier, or under a binocular microscope of the Greenough type carrying
similarly magnifying lenses. Material to be removed can be exactly lo-
cated in the parent colony. It is found possible in dealing with most
Aspergilli (1) to remove conidia from a single head, (2) to remove one or
more heads borne upon a single hypha at the margin of the colony, or (3)
to select vegetative hyphal tips from a single mycelial sector. It is usually
possible to avoid (1) heads and conidia of other species or strains, (2) foreign
mycelia, and (3) bacterial contaminations present in the substratum.
Purity in repeated culture over many years has been possible by this pro-
cedure.

h

B

Fig. 10. Single and three-point cultures of Aspergillus foetidus on Czapek's solu-
tion agar, room temperature, 10 days, X i approximately. Note that spore produc-
tion extends to the colony margins in the central triangle of 'figure B, and almost to
the outer colony margins as well. In figure A, sporulation is limited to the central
area only. Figure B is favorable for direct examination with the low power objective
of the compound microscope; figure A is not.

Dilution Cultures

If dilution cultures are to be used, the suspension of conidia should be
sufficiently dilute so that the individual petri dish will show not more than
six, but preferably not over three, colonies. Such dilutions are difficult to
gauge and often result in some plates crowded with numerous colonies while
others contain none. The colony locations are not under control. In
general, the dilution of spores in successive water blanks with the subse-
quent plating of aliquots from such dilutions has not been found sat-
isfactory.

An alternate technique involves the introduction of one loopful of inocu-
lating material into a tube of melted agar which is then rolled or shaken,

CULTIVATION AND EXAMINATION 41

and a loopful removed to a second tube, followed by the same manipulation
a third or fourth time. The higher dilutions are then poured into petri
dishes where colonies develop. This method has all the faults of the water
blank dilution technique. The procedure has been widely used but has not
been followed in our study of the Aspergilli. Blakeslee (1915) and other
mycologists have prepared such dilutions in bottles by slowly rotating the
bottle in a cold water or ice bath, thus allowing the agar to congeal in a thin
layer against the glass surface. This practice has little to recommend it:
(1) the resulting colonies could be isolated more readily from a petri dish,
and (2) it is basically impractical to make satisfactory observations regard-
ing the character and structure of a mold colony, or the gross features of its
fruiting structures, when observations have to be made through the curved
and non-uniform walls of a glass bottle.

Smears and Streak Cultures

The practice of smearing a suspension of mold spores over the whole plate,
or the whole length of a slanted agar tube, results in a growth of mycelium
which covers the entire surface and usually produces a greater mass of co-
nidia. But individual colonies are usually unidentifiable in such preparations,
hence they are of little value in providing those critical details of colony
habit, coloration, and texture necessary for identification and classification.
In general, mycelium derived from different conidia of the same strain will
intertwine without inhibition, and even anastomose, if they come into con-
tact in the early stages of growth; for example, before any sign of conidium
production appears. If the spacing of the spores is great enough to permit
the establishment of small fruiting colonies before such contact is made, the
colonies frequently do not converge and complete coverage of the surface of
the agar with the production of maximal quantities of conidia does not oc-
cur. Closely-placed seeding of spores is useful to obtain the largest possible
supply of conidia; but to study the normal characters of a species, individual
colonies must be allowed to develop without interference by others of the
same or different species.

Streak cultures can be employed to advantage in freeing one mold from
another, or from other contaminating organisms. By touching a sterile,
moistened needle to a single conidial head or other selected fruiting surface
of limited extent, and subsequently streaking this repeatedly across the
surface of an agar plate, isolated colonies of the desired species can usually
be obtained. Occasionally it is desirable to streak two plates in succession
in order to secure a satisfactory separation of colonies. In any case the
spore source should be selected with great care and the use of a low-power
binocular microscope or pocket magnifier is recommended. When this
method is employed, it is desirable to reisolate from these individual colo-

42 A MANUAL OF THE ASPERGILLI

nies as soon as possible after sporulation begins. For freeing one species or
strain of Aspergillus from another, or from a contaminating Penicillium,
streaking upon Czapek's solution agar usually gives satisfactory results.
The method finds its greatest usefulness in separating comparatively slow-
growing molds, such as the Aspergilli, from such very rapidly growing forms
as the Mucoraceae, Trichodcrma, etc. The critical step in this procedure
is to isolate the slow growing Aspergilli during the first two to three days
before the whole plate is overrun by the spreading, faster growing forms.
If the contamination is bacterial in nature, malt extract agar or some other
medium of more acid reaction should be substituted. Diluting the spores
in water before streaking is not recommended since this often tends to dis-
perse the contaminating organism more than it does the desired form; this
is especially true with bacterial contaminations, actinomycetes, and other
minute forms.

Single Spore Cultures

The isolation of cultures from single spores is a time-honored technique
with many mycologists, and with many workers it is considered a "must."
When properly employed, there is much to recommend this practice. There
are also certain dangers inherent in this procedure, hence we believe it worth
while to consider the subject at some length and to analyze both its advan-
tages and its limitations. As generally employed, the primary objective of
single spore isolations is to secure cultures of unquestionable purity. If the
operation is skillfully performed and adequately verified, there can be no
doubt but that the resulting colony will represent a pure culture of a single
species or strain. The continued purity and physiological stability of such
cultures, however, cannot be taken for granted. There is no substitute for
vigilance, and the culture thus isolated must be kept under critical ob-
servation.

As a means of purifying a culture, i.e., separating it from foreign forms,
the single spore method undoubtedly has its place ; but it cannot be recom-
mended as a means of preserving the morphological and physiological sta-
bility of a particular strain. One has only to study the reports of Stakman
and his associates to realize that monospore selection and isolation is not a
touchstone to strain stability. In fact, they have ably exploited the tech-
nique to show just the opposite. While we do not have an accumulation of
data on the Aspergilli and related genera which can compare with that on
the smuts and other pathogenic fungi, we do have enough information to
know that one of the best methods of obtaining change in some of the
Aspergilli is to make a series of successive monosporous isolations. For
every species and strain there is apparently a normal range of natural
variation, and by isolating single spores it is often possible to secure certain

CULTIVATION AND EXAMINATION 43

progeny which represent essentially the outside limits of such variation.
This is worth while since it offers one means of securing strains which may
prove more useful than the parent culture for some particular purpose,
industrial or otherwise. The single spore method can be of real value in
purifying a culture. It can be of real value as a means of "dissecting" a
culture. But once a promising strain has been discovered and its purity is
established, perpetuation by the transfer of masses of conidia is the best
safe-guard for preserving, in a constant condition, its morphological and
physiological characteristics.

Some of the techniques by which such single spore isolations can be made
follow :

(1) Serial Dilation: Spores are thoroughly suspended in water and the
resulting suspension is subsequently diluted in sterile water blanks in steps
of 1:5, or 1 :10. One cubic centimeter aliquots from two or three selected
dilutions, depending upon the density of the original suspension, are added
to sterile petri dishes. Melted agar at approximately 45° C. is then added
to these plates, which are rotated to secure uniform mixing of the still liquid
agar and the diluted spore suspension. The plates are then incubated at
room temperature and isolates are made from plates showing a limited num-
ber of colonies which are uniformly separated. Using this technique, one
cannot be certain that any particular colony results from a single spore, but
if the original suspension was properly prepared, one can feel sure that more
then 95 percent of the colonies resulted from single spores. More uniform
suspensions of spores can be obtained by adding to the suspending medium
some suitable detergent or aerosol. For this purpose we have successfully
used sodium lauryl sulfonate in concentrations of 1 : 10,000 or 1 : 100,000
without apparent harmful effect upon the molds under study. The dilu-
tion method of securing "single spore" isolations is more rapid than any
other, and for many purposes it is quite satisfactory.

(2) Selection and Removal of Individual Spores: Where the investigator
wishes to be positive that every colony results from a single spore, it is
necessary to employ some technique combining actual microscopic examina-
tion with some device for the mechanical removal of selected spores. The
various types of micro-manipulators are well-suited for this work and, with
sufficient practice, single spore isolations can be made quite satisfactorily
with any of these. Dilute spore suspensions in water are mounted on the
undersurface of a cover slip supported by a glass chamber open at either
one or both ends. With the aid of mechanical controls and a micro-pipette,
a single spore is withdrawn from the suspension and ejected upon a suitable
substratum where the spore develops into a mature colony.

Single spores can likewise be removed by mechanical cutting devices
such as those described by LaRue (1920), Keitt (1915), and Lambert (1939).

44 A MANUAL OF THE ASPERGILLI

A comparatively thin spore suspension is spread on an agar surface, and
single well-separated spores are located with the microscope. A small
cutting device, mounted on a holder screwed into the nose-piece of the same
microscope, is lowered into the agar and a small block bearing the selected
spore is removed. This is subsequently transferred to a suitable culture
surface where the colony develops.

At the Northern Regional Research Laboratory we have employed a
somewhat different and simpler method. A thin spore suspension is spread
evenly over the surface of a firm agar gel that has been specially filtered to
remove all particulate matter. This is incubated overnight and the spores
allowed to germinate. On the following day, well-separated sporelings are

Fig. 11. Single spore isolation. A, A single, well-isolated, germinated conidium
of Aspergillus niger. B, The same removed on a small agar block and transplanted
to a fresh agar plate as described in the text, p. 44, X 300.

located with the aid of a microscope and their positions marked on the
under-surface of the culture dish. These are then checked with a 8 mm.
objective and 10 X or 15 X oculars to insure that there are no other un-
germinated spores in the same area. Using a wide-field binocular of the
Greenough type and very small micro-scalpels fashioned out of platinum-
iridium wire (B and S gauge 22 or 24), small agar blocks on which the spores
spores rest are transplanted to fresh agar plates. Each of these agar blocks
is then examined again with the 8 mm. objective to insure that the selected
spore has been transplanted. An experienced worker can isolate from 25
to 30 spores within a period of an hour. Photographs showing essential
steps in this technique are presented in figure 1 1 .

CULTIVATION AND EXAMINATION 45

Hanging Drop Cultures

The drop of culture fluid inoculated with conidia and hanging from a slide
or cover glass into a closed chamber can be incubated and examined readily.
It furnishes information as to the percentage of conidia which are viable,
the changes which occur in the germinating spore, such as swelling, bursting
along definite lines, germination from specialized germ-pores, or branching
of the germ tube, but descriptions of fruiting structures in such hanging
drops are worthless in the study of the mold colony or normal fruiting habits
of the species.

TEMPERATURE

The great majority of Aspergilli grow well and sporulate abundantly at
temperatures of 23° to 26° C. For this reason, most cultures can be incu-
bated on laboratory tables or shelves, and it is not necessary to give special
consideration to incubation. There are, however, certain exceptions.
The large-spored members of the Aspergillus glaucus group, such as A . echin-
ulatus and A . niveo-glaucus, grow more rapidly and fruit more abundantly
at 20° than at 24° to 25° C. (Thorn and Raper, 1941). This temperature
response is especially marked in A. medius: at 18° to 20° C. growth is rapid,
abundant large conidial heads are produced, and numerous perithecia are
developed ; at 25° C. and above, growth is restricted, few and smaller conidial
heads are developed, and only occasionally perithecia are produced (fig. 12).
On the other hand, the very abundant small-spored members of the A.
glaucus group, such as A. re-pens, A. chevalieri, and A. amstelodami, grow
rapidly and fruit abundantly at 30° C. (Thorn and Raper, 1941). In A.
janus (Raper and Thorn, 1944), two different types of conidial heads are
produced and the ratio of these types is strongly influenced by temperature
(fig. 12): at 18° to 20° C. almost all heads are white with clavate vesicles
and are borne upon long conidiophores ; at 30° C. almost all heads are dark
green with globose vesicles and are borne upon short conidiophores (see
species description, p. 187).

When grown upon suitable media and incubated at 20° C. in the presence
of light or alternate light and darkness, Aspergillus giganteus produces
large heads on long conidiophores ranging up to 5 or even 10 cm. In similar
cultures incubated at 30° C. heads are smaller and conidiophores are uni-
formly short, rarely exceeding 1 cm. in length, whether the cultures are ex-
posed to light or incubated in total darkness. A. terreus, A. carneus, and
A. fischeri thrive at temperatures up to 35° C, while A. fumigatus grows
well at 45° or even 50° C. In all of these forms growth is more rapid and
sporulation more abundant at 30° C. than at normal laboratory tempera-

46

A MANUAL OF THE ASPERGILLI

Fig. 12. Influence of temperature upon growth and sporulation in Aspergillus
medius, NRRL No. 124, and Aspergillus janus, NRRL No. 1787. A , B, and C, Asper-
gillus medius, 17 days at 12° C, 20° to 22° C, and30° C, respectively ; note maximum
growth and limited sporulation at 20° to 22° C. D, E, and F, Aspergillus janus, 3
weeks at 20° C, 24° to 25° C, and 30° C, respectively; note evidence of white heads
only at 20° C, abundance of white and presence of green heads at 24° to 25° C, and
green heads only at 30° C. Temperature is the only variable.

CULTIVATION AND EXAMINATION 47

tures of 24° to 25° C, although wholly typical cultures are produced at
both temperature levels.

In the production of spores for biochemical investigation (see p. 37) and
in the conduct of such studies themselves, temperature is often very im-
portant. In work of this kind, it is essential to determine the optimum
temperature for each organism and process, and then to control this within
a narrow range in order to secure consistent and reproducible results.

Temperature is known to be a critical or limiting factor only in a few
species, but its effect in these is sufficiently pronounced that its influence
should not be disregarded in any case.

CULTURE TOOLS AND EQUIPMENT

Transfer Needles

In making transfers, we have used with satisfaction for many years a
Xo. 20 or No. 22 B. and S. gauge nichrome wire thrust into a slender brass
or stainless steel tube so that only about 15 to 20 mm. are exposed. (The
tubing employed is of the type used in temperature controls of mechanical
refrigerators and other thermostatically controlled devices.) The point of
this wire is then ground down to the sharpness and smoothness of a needle.
This instrument can be heated to redness in the flame a great many times
with only the occasional necessity of resharpening. It thus has many ad-
vantages beside cheapness: it is firmer and takes a better point than plati-
num; it withstands sterilization in the flame, which promptly destroys the
usefulness of steel ; and it can be made any size or length to suit the work-
man's purposes or preferences.

Loops

Loops of various dimensions are very useful and can be inserted into
handles made from small brass or stainless steel tubing as noted above.
These, too, can be fashioned from nichrome wire, but it has been our
experience that loops made of platinum-iridium wire possets certain marked
advantages. While these are not as rigid as nichrome loops, they are much
more rigid than loops of pure platinum, and they can be re-heated indefi-
nitely without corroding. Loops of this type will be found especially
useful in making mass inoculations, such as the seeding of large tubes and
plates for the production of spores to be used in various types of experi-
mental work.

Mounting Fluid

In the microscopic examination of the Aspergilli it is often satisfactory
to mount conidia, heads, or other structures in water. It is more generally

48 A MANUAL OF THE ASPERGILLI

satisfactory, however, to use some mounting fluid of a composition designed
neither to swell nor plasmolyze the tissues to be observed. Such a
mounting fluid was developed by Amann as early as 1896 and has been
used by mycologists, quite generally, for many years. Its composition
is as follows:

Carbolic Acid Crystals (c.p.) 20.0 grams

Lactic Acid (sp. gr. 1.2) 20.0 grams

Glycerine (sp. gr. 1.25) 40.0 grams

Distilled water 20.0 cc.

The carbolic acid crystals are first liquefied by heating in a water bath.

The mounting fluid is normally used without the addition of any dye, since
in the diagnosis of the Aspergilli the natural colors of conidiophores, conidia,
etc., are very important. If it is considered desirable to stain the tissues
under observation, it is possible to incorporate into the lactophenol solution
some coloring substance such as cotton blue, eosin, or some other aniline
dye. In making mounts of the Aspergilli, it is profitable to wet the material
first with 70 percent alcohol to drive off air bubbles, and then quickly add a
small drop of the lactophenol prior to the placement of the cover glass.

Incubators

Almost all of the Aspergilli grow well and sporulate abundantly at
laboratory temperature. There are, however, certain exceptions to this
general rule (see p. 45), and for this reason it is desirable to have available
an incubator which can be regulated at temperatures below that of the
laboratory and others covering different ranges up to 37° C, or even 50° C.
The type of incubator is not critical and almost any type of cupboard, or
room, will prove satisfactory if the temperature can be controlled to
within 1° C. ±, and if the air is neither excessively dry nor humid to the
point where cotton plugs become moist upon continued exposure.

Microscopes

In the study and identification of Aspergilli it is essential to have at one's
disposal a good-quality compound microscope. If possible, this should be
provided with apochromatic lenses. In our experience we have found a 3
mm. objective used in conjunction with either 10 X or 15 X oculars to give
us magnifications and a degree of definition which is most satisfactory for
the examination of conidial structures. While it is not absolutely neces-
sary, it is desirable to have, in addition, a low-power, wide-field dissecting
microscope covering magnifications from 10 to 40 diameters. In the ab-
sence of such a microscope a high-quality pocket magnifier provides a good
substitute.

CULTIVATION AND EXAMINATION 49

Photographic Equipment

A limited amount of photographic equipment is a very valuable aid in
the study of this or any other group of molds. A camera should be avail-
able which can be used in conjunction with a compound microscope, and
to which low-power lenses can be attached directly. In addition to a good-
quality lens producing pictures above and below natural size, a series of
Tessar lenses which will provide magnifications of from 5 to 30 diameters
is extremely useful. With the aid of these, details of colony structure can
often be recorded which cannot be pictured with the lenses of the compound
microscope and which are very difficult to describe adequately in words.
With these low-power lenses it is possible to photograph types of fruiting
heads, the relative abundance of conidial structures to vegetative mycelium,
and the relationship between each of the above and sclerotia or perithecia
when such structures are present.

Chapter V
PRESERVATION OF CULTURES

Any mold that is valuable because it has been used in fundamental
research work, because it has been found useful in some industrial process,
or because it is a significant agent in some destructive or pathogenic situa-
tion should be preserved to insure its identity for subsequent use or refer-
ence. Identification by description will take the careful worker to the
group, species, and often to the variety as based upon morphology or some
conspicuous character, but the reidentification from description of the
exact organism used in a biochemical investigation or discovered in some
ecological situation is generally impossible. Culture collections have,
therefore, developed. The Centraalbureau voor Schimmelcultures (cited
by them as C.B.S.) at Baarn, Holland, the National Type Culture Collec-
tion in London, and the American Type Culture Collection at Washington
are well-known sources of such material. More recently established, but
containing a greater number of industrially important molds, is the culture
collection of the Northern Regional Research Laboratory (commonly
cited as N.R.R.L.) at Peoria, Illinois. The Aspergilli are especially well
represented, and contained in the collection are almost all of the species
considered in this manual, together with records which check the identify-
ing numbers of this collection against the records of the source collections*
which were brought into it.

In undertaking a comparative study of the Aspergilli, or any other group
of molds, it is essential to maintain in a viable state a large number of
isolates representing diverse species and strains. By this means, it is often
possible to interpret current isolates and accessions in terms of historic
types and concepts. In maintaining such a collection of molds, the objec-
tive should be to preserve viability without growth or germination of
spores during the storage period. By so doing, the user can reasonably
expect to maintain his organism without variation, degeneration, or muta-
tion. A number of techniques can be employed to advantage, the more
useful of which will be considered in some detail with certain of their
advantages and limitations noted.

* Currently contained in this collection are the molds formerly maintained by
Thorn and Church, and later by Thorn and Raper, in the United States Department
of Agriculture, Washington, D. C; many of the Mucorineae maintained by Dr. A. F.
Blakeslee for many years at the Carnegie Institution, Cold Spring Harbor, Long
Island, New York; a large number of miscellaneous forms from the Harvard Univer-
sity Collection initiated by Prof. R. Thaxter and more recently maintained by Dr.
D. H. Linder; and limited numbers of cultures from many other collaborators.

50

PRESERVATION OF CULTURES 51

A gar-Slant Method

The method most generally employed for maintaining mold cultures,
and the one which has been successfully used by the writers for many years,
may be termed the agar-slant method. This involves the periodic transfer
of spores from old agar slants, or plate cultures, to new agar tubes. The
composition of the substratum is varied to suit particular requirements and
groups of organisms. In our work we regularly employ the Czapek's
solution agar. Few, if any, strains make their maximum growth on this
medium, but it has been our experience that they maintain exceptionally
well any characteristic morphological or physiological features which may
characterize them. It is necessary to know the expected viability of all
cultures to be maintained, and to gauge the intervals of transfer accordingly.
With the Aspergilli, transfer every 8 to 9 months is sufficiently frequent for
all species, with the possible exceptions of A. citrisporns and A. itaconicus,
and a period of one year is not too long for most forms. In practice it is
advisable to handle separately the few very short-lived species, and to set
the regular period of transfer well within the known viability period of the
remaining forms. Transfer of the general collection at least once each year
insures a complete survey, within the yearly period, of all strains main-
tained. New tubes are inoculated at least in duplicate, while triplicate
preparations afford a desirable margin of safety. The new cultures are
incubated for 2 to 3 weeks, or until a good crop of spores has developed.
Incubation at room temperature is suitable for most of the Aspergilli,
although a few forms such as the large-spored members of the A. glaucus
group sporulate more abundantly at 20° C. The correctness of the cultures
is then checked with a wide-field binocular or a 10 X pocket magnifier, the
plugs are poisoned to preclude any possibility of subsequent contamination,
and selected tubes are placed in storage. Cultures can be stored for
reasonable periods at room temperature; certain species will remain viable
for many years at 24° to 26° C. The viability of most species is materially
lengthened and the possibility of progressive variation reduced by storage
at 2° to 4° C. (i.e., above any danger of actual freezing but sufficiently lowr
to prohibit further growth and possible dissociation). Tubes of any de-
sired size may be employed. We have found lipless tubes 15 by 125 mm.
to be quite satisfactory since they provide adequate culture surface and at
the same time require much less storage space than the larger tubes com-
monly in use. Each culture should be maintained at least in duplicate,
with the different tubes of each pair stored in separate refrigerators. With
the accidents and failures of refrigeration, the possible escape of toxic gases,
or the possible ingress of contaminations that escape the usual inspection,
the maintenance of not less than two complete series of strains is a necessary
precaution. If natural conditions, such as temperature and relative hu-

52

A MANUAL OF THE ASPERGILLI

midity, are favorable as at Baarn, Holland, refrigeration may be dispensed
with, but watchfulness against invasion by mites becomes more important.
Additional slants should be prepared for cultures which are frequently used.
Agar slant cultures are convenient for use, easily examined and com-
pared, and easily replaced (fig. 13 A). They are, however, easily contami-
nated when handled carelessly. Uneven drying subjects the culture to

Fig. 13. Methods of maintaining stock cultures as discussed in the text. A,
Cultures growing on agar slants. B, Cultures preserved in lyophile form; note the
compact, chalky pellets formed by the dried serum in which the spores are suspended.
C, Cultures preserved in dry soil.

extremes of contrast in concentration of media and metabolic end products,
between the thin edge of the slant and the heavier mass in the bottom of the
tube. Variations (apparent or real) often appear in the stored cultures
and these may be propagated in subsequent transfers. As a safeguard,
stock cultures should be grown in petri dishes after several transfers in tubes,
thus making possible more complete examination to maintain purity and
typical morphology.

PRESERVATION OF CULTURES 53

Preservation in Lyophile Form

Studies now in progress at the Northern Regional Research Laboratory
indicate that many, if not all, of the Aspergilli can be successfully main-
tained in a dried state for extended periods. Viability tests for a number of
species including A. terreus, A. niger, A. oryzae, A. flavus, and A. itaconicus
have been made at 3£ years; while a much greater and wider variety of
forms has been tested at 20 to 24 months. Positive results have been ob-
tained with all cultures tested, although comparatively few colonies devel-
oped from certain strains of A. niger, A. flavus, and the large-spored
members of the A. glaucus group. Observations are being continued and
in time information will be obtained as to the feasibility of employing this
as the principal means of maintaining a collection of molds. It is known
that many bacteria, especially staphylococci, streptococci, and pneumo-
cocci can be successfully preserved for periods up to 16 to 18 years (Elser,
Thomas, and Steffen, 1935; Swift, 1937). Wickerham and Andreasen
(1942) have presented evidence covering a period of one year which suggests
the practicability of applying the method to the yeasts. Such information
as we have to date regarding the molds seems to indicate that the method
may prove of great significance in two ways: first, as a means of prolonging
viability, and second, as a means of preserving in viable form spores of a
particular "generation," or other selected origin, which can be used in
comparative tests over a period of many months or even years.

The drying technique employed at the Northern Regional Research Lab-
oratory is essentially like that described by Wickerham and Andreasen
(1942) and may be briefly summarized as follows:

Employing aseptic techniques throughout, the spores from selected
cultures are suspended in sterile beef, or horse serum. The resulting sus-
pension is then dispensed into small cotton-stoppered Pyrex glass tubes
6 mm. by 100 mm. that have been properly labeled with glass-marking ink.
Approximately 0.05 to 0.1 cc. of the spore suspension is added to each tube
by means of a long thin-necked pipette. Most of the cotton plug is burned
away, and the remaining portion pushed down into the tube to prevent
possible contamination during the drying process. The tubes are inserted
in rubber sleeves on the manifold, as shown in figures 14 A and B, and low-
ered into a freezing bath of carbon dioxide ice and methyl cellosolve at a
temperature of approximately —40° C. The suspension is frozen almost
instantaneously. The manifold is connected to a vacuum pump and evacu-
ation and desiccation initiated. Water is removed from the system by the
insertion of a water-trap immersed in a C02-methyl cellosolve filled Dewar
flask as shown in figure 14A, or in a column of drierite (anhydrous CaS04)
as shown in figure 14B. After a few minutes the temperature of the bath

54

A MANUAL OF THE ASPERGILLI

Fig. 14. Apparatus employed at the Northern Regional Research Laboratory for
preserving microorganisms in lyophile form. A , Table model of thirty tube capacity,
utilizing a trap immersed in a Dewar flask filled with CO2 ice and methyl cellosolve
to collect the water vapor removed from the drying preparations. B, Larger, self-

PRESERVATION OF CULTURES 55

surrounding the tubes is raised to approximately —5° C.1 by supplying
additional methyl cellosolve that has been precooled to approximately 0° C.
The bath is maintained at this level continuously until the preparations
appear thoroughly dry. In drying, the serum contracts slightly to form a
well-defined chalky pellet. When the pellets are apparently dry, the tubes
are raised above the bath and evacuation is continued for one-half to three-
quarters of an hour at room temperature to insure as complete removal of
water as possible, after which time they are sealed off with a gas-oxygen
torch (fig. 14 A). On the following day each tube is tested for the presence
of a good vacuum by means of a high-frequency, spark coil tester, and only
those tubes which show such a vacuum are retained. Tubes not maintain-
ing a satisfactory vacuum are very rarely encountered in actual practice.
Quadruplicate tubes are regularly made for each stock culture, and the
finished preparations are stored in a refrigerator.

In recultivating the molds, the tubes are marked with a file scratch, sur-
face-sterilized, and the tube broken inside a wrapping of sterile cotton.
The content, which is in the form of a well-formed pellet (fig. 13 B), is then
dissolved in 1 to 2 cc. of sterile broth or water. This is streaked on agar
plates and colonies are allowed to develop. New isolations can be made
within a period of a few days. It is possible, of course, to go directly from
the lyophile tubes into flasks or other cultures used in actual experiments,
but generally speaking, much larger quantities of material would need to
be processed.

The feasibility of preserving molds in lyophile form over long periods
has by no means been proved, but results to date are very encouraging.
Should it be found that spores of molds, like bacterial cells, can be kept
viable by this method for many years, it will prove ideal as a means of pre-

1 Wickerham and Andreasen in 1942 governed the temperature at which the sus-
pension was dried by adjusting the level of the tubes above a bath which was kept
at a very low temperature. Subsequent to this, Wickerham developed the procedure
outlined above.

contained and portable unit of sixty tube capacity (designed by Dr. L. J. Wickerham)
which utilizes a column of anhydrous calcium sulphate ("drierite") to collect water
vapor removed from drying preparations. Ai and B\, Glass manifolds; A2 and B2,
Thermometers; A3, Dewar flask containing water vapor trap immersed in CO2
ice and methyl cellosolve; Bu Column of drierite; A^ and Bb, Freezing bath con-
taining CO2 ice and methyl cellosolve in which preparations are immersed for
quick freezing and subsequent temperature control; Ae, and Be, Vacuum pump; At,
Vacuum guage (mounted at opposite end of apparatus shown in figure B) ; As and Bs,
Gas-oxygen torch for sealing off dried preparations; B9, Terminals on which finished
tubes are mounted to be tested for presence of good vacuum by means of a high fre-
quency spark coil tester (not shown); Bio, Oxygen tank; Bu, Screw lift for raising
and lowering manifold and attached tubes (In figure A , manifold is raised and lowered
manually and locked into position by means of a wing bolt).

50 A MANUAL OF THE ASPERGILLI

serving large culture collections. It possesses certain marked advan-
tages :

(1) There is no possibility of contaminants entering the sealed prepara-
tions.

(2) The investigator recultivating the molds starts with the actual spores
contained in the original suspension.

(3) The space required for storage of a large number of lyophile prepa-
rations is much less than for any other type of culture (fig. 13 B).

Preservation in Soil

Soil has been successfully employed as a means of preserving vigorous
stock cultures over long periods. As early as 1918, Barthel (Cent. Bakt.
II, 48: 340-49. 1918) reported the successful maintenance of yeasts and
bacteria in this medium and modifications of this technique are now em-
ployed in many laboratories for preserving bacteria. Greene and Fred in
1934 compared cultures of various molds preserved for two years in soil
with the same strains continuously maintained on malt extract and malt
extract-potato-glucose agars and on bread. In their experience, soil prepa-
rations were most satisfactory, and Professor Elizabeth McCoy (personal
communication) has recently reported cultures of A. sydowi preserved in
this manner to be viable after nine years. Since the publication of Greene
and Fred's work, the soil method has been rather generally used by the
Wisconsin group as a means of preserving valuable stock strains of molds.
Furthermore, it is known to have been successfully employed during the
past two years by a number of laboratories to maintain cultures of penicillin-
producing molds in a high and uniform state of productivity. The soil
substrate used by Greene and Fred was prepared as follows:

"To air-dried orchard loam soil (Miami silt loam) sufficient water is added to
bring it to a moisture content of about 20 percent. The soil is then transferred in
convenient amounts (about 5 grams on a dry basis) to ordinary half-inch (1.27 cm.)
culture tubes. The tubes are plugged with cotton and given four 3-hour steriliza-
tions at 15 pounds per square inch (1 kg. per sq. cm.) pressure on alternate days, and
tested for sterility by addition of yeast-water-glucose broth to tubes selected at
random. The tubes are then inoculated with 1 cc. of a heavy spore or mycelium
suspension of the desired mold and kept at room temperature. That there is appre-
ciable growth and sporulation on the soil can usually be ascertained without difficulty
by direct microscopic observation. While the addition of nutrient to the soil may
bring about somewhat greater growth, it does not seem to enhance the keeping quali-
ties of the cultures.

"It has been found possible to preserve on soil mold stocks used for large-scale
growth— namely, Aspergillus fischeri, A. sijdowi, and Penicillium chrysogenum— for
over 2 years without loss of their essential and desirable characters. . . . Moreover,
the gross colony characters have remained much more constant than did those of the
corresponding cultures maintained in the usual way on agar slants. The soil cultures

PRESERVATION OF CULTURES 57

can be recovered as required simply by streaking some of the soil particles on fresh
agar slants.

"It is not in all cases advisable to depend on soil alone for the preservation of
valuable stocks, but reserve stocks may without difficulty be prepared on soil, and
the writers believe that in many instances soil will be found to be an excellent medium
for maintenance, with a minimum of change over long periods of time."

During the past two years the soil method has been used at the Northern
Regional Research Laboratory with but minor modifications of the tech-
nique cited above (fig. 13 C). Its principal advantages lie in the fact that
(1) the viability of strains is apparently lengthened, and (2) from a single
stock tube, opened with proper care, repeated cultures can be started
simply by removing some of the soil particles to suitable substrata.

Vegetable Substrata

The oriental fermentation industries maintained their inoculating mate-
rial as selected rice or soybeans upon which the mold had been grown under
favorable conditions to produce maximum quantities of spores. This nu-
trient, dried and packaged, was stored and sold under the Japanese name
"Koji". Samples examined after several years showed excellent viability.

Bainier was a pharmacist. He distributed licorice root in sections 5 to 10
mm. in diameter and 5 to 8 cm. in length in test tubes, sterilized them, and
kept his cultures regularly for years upon them. Tested by us, the method
was a very satisfactory laboratory practice. American mycologists have
successfully used bean stems for the purpose. Apparently any organic
material which provides frameworks of cellulose enmeshing sufficient nutri-
ents to support mold growth without complete breakdown of the mass may
be used.

CONTAMINATION

Mixed Strains

In the routine conduct of cultural work, contaminations of cultures of one
species of Aspergillus by other species, or species of other genera, is very
common. The conidia of most molds are exceedingly light and are carried
freely in the air. Entire exclusion of such contamination is difficult. In
dealing with contaminations, several problems arise and different proce-
dures are possible. A colony of a single Aspergillus, well established,
usually inhibits the growth of other species developing in the immediate
vicinity. Even if invasion occurs, the effects are commonly so distinct as
to leave little doubt as to the limits of the different forms. When, however,
the contamination with spores or mycelium is carried in the inoculum and
so placed as to germinate in intimate contact with the organism desired, (1)
the species may sector out, and hence be easily recognized, or (2) the colony

58 A MANUAL OF THE ASPERGILLI

resulting may assume the character of either organism with the other

present only as an inconspicuous, even unrecognizable, mycelium with

dwarfed heads, yet continue present for a long period. In this way the

retarded species may suddenly reappear in some later transfer upon media

favorable to it. In other cases, the dominant species may grow and fruit

in an erratic manner that is deceptive in suggesting a reaction to the medium

r other physical factors. Again, in less common cases, the two may grow

jid fruit together without apparent inhibiting effects. This is the most

lifficult form of mixture.

Mixtures are sometimes encountered in which mycelia, sterile under all
conditions tested, become so intimately mixed with the mycelium of an
Aspergillus or Penicillium as to persist through many generations without
apparent effect in the earlier states of growth of the Aspergillus, but develop
as overgrowths of sterile hyphae in very old cultures. Many of these forms
can be isolated, but they defy identification because of an absence of diag-
nostic characters. Such sterile mycelia may arise from the species studied,
but unless such origin can be definitely proved, they must be regarded as
contaminants. Great care is necessary in interpreting cultures producing
sterile overgrowths, since the contaminating organism, or non-sporulating
variant of the same strain, may induce marked changes in the physiological
activity of the species supposedly pure.

Secondary Growth

In many species, part or all of the conidia produced by a colony germinate
and cover the primary mycelium with more or less abortive hyphal growths,
many of them unrecognizable unless traced by their origin. Again spores
floating on the surface of a globule of transpired fluid may germinate, their
hyphae interlace and a hollow ball surrounded by felted mycelium produce
structures which probably account for reported perithecia without asco-
spores. Such overgrowths, being irregularly produced, interfere with one's
judgment as to the whole character of the colony.

Replacement by Other Species

Other species of fungi invade mold cultures and some of them become so
intimately associated with the mycelium and conidia of particular species
that it is difficult to eliminate them by the ordinary method of transferring
spores with a loop or wire to streaks or stabs. Proper dilution culture
presents the possibility of elimination but demands careful examination
and selection from the resulting colonies. The progressive replacement of
a particular species, by invading organisms, is constantly encountered in
examining cultures passed from laboratory to laboratory. The original
organism may disappear without detection if the displacing species bears

PRESERVATION OF CULTURES 59

a superficial resemblance to it, or if no detailed examination of successive
transfers is made by a worker really familiar with the specific characters of
the stock strain. The physiological or biochemical investigator, obtaining
such a culture assumed as correctly named, will be seriously misled in the
interpretation of his experimental results. No culture should be used for
such an investigation without being fully identified at the outset and having
its proper appearance and reactions sufficiently studied to insure the relia-
bility of the results during the progress of the work. In other words, before
undertaking to do biochemical or physiological work, sound scholarship
calls for adequate precautions in the study and identification of the original
material supplemented by such mastery of its morphology and variability
as will insure its maintenance in proper condition.

Mold Disease of A . niger

A mold disease of Aspergillus niger is common in cultural study. The
colonies of the Aspergillus are overrun with an olive-green Penicillium
belonging in the Biverticillium group close to Penicillium rugulosum Thom.
This mold invades the mycelial felt, winds its hyphae within and about the
conidiophores, and fruits in a radiating series of short-stalked penicilli
surrounding the heads and upper halves of the conidiophores. This is
beautifully illustrated in figure 15, made by Edward Yuill and sent to us by
his brother John L. Yuill of Yorkshire, England. The black fruiting surface
may be completely covered with the olive-green conidial masses of the
Penicillium. If A. niger is grown in trays for acid formation, spots infected
in this way may be killed and disintegrated, thus interfering with fermenta-
tion, while the infected areas may be seen to drop out when the blanket or
felt of A. niger is lifted from the surface of the liquid. Other species when
inoculated have been irregularly affected, some strains of the same species
were attacked, whereas others were apparently immune.

Bacteria

Freedom from bacteria is essential to uniformity in the appearance and
in the reactions of molds. Colonies infected with bacteria may be un-
changed in character, but usually show marked physiological differences
when compared with colonies free from contamination. Associative action
may have important effects upon both organisms. Sartory (1920) dis-
cussed without identifying a species producing ascospores, but only when
accompanied by a particular bacterial associate. Nevertheless, a symbiotic
colony must not be allowed to masquerade as a pure culture representative
of a species. The next contaminated colony of the same species of mold
but with different bacteria may present a very different picture. These
conditions have been met often enough to make the emphasis upon freedom
from bacteria essential in study of this group.

60

A MANUAL OF THE ASPERGILLI

Mites

Mites are very common in rotting vegetables, especially in partly dried
condition, in dried meat products, in hard cheeses, and in organic soil
masses. They are thus common associates of molds as they occur in nature,

Fig. 15. Penicillium sp. parasitic on Aspergillus niger, X 165. (Photograph by

Edward Yuill)

hence the worker who handles moldy substrata, in isolating his organisms
must constantly watch for them. In size, mites are commonly just about at
the limit of visibility by the unaided eye. One accustomed to them will
detect them readily; but until seen and the appearance of their depredations

PRESERVATION OF CULTURES 61

understood, they can pass unnoticed for considerable periods by persons
who are otherwise good culture workers. Mites will crawl from petri dish
to petri dish, leaving behind them a trail of bacterial and mold contamina-
tions, as well as streams of eggs which develop rapidly into more mites that
actually destroy the colonies. Since mites have preferences as to food,
some species are invaded and others avoided. To reach an attractive food
supply a mite will frequently go through a cotton plug as ordinarily made
and occasionally seems to get through even a paraffined plug. As a factor
in the mixing of strains of molds in a laboratory collection, mites must not
be ignored.

Similar mixing and contamination occurs frequently whenever a labora-
tory becomes infested with ants, roaches, or other insects.

Poisoning Cotton Plugs

Mites: Entire elimination of mites by sanitary measures is possible but
often not attained. As a precaution in the preservation of stock cultures,
some scheme of poisoning should be used. One of these formulas consists
of dipping the tips of the cotton plugs in a solution of the following com-
position:

95% alcohol 95 cc.

Bichloride of mercury 0.5 gm.

Glycerine 5 cc .

Color with any aniline dye.

Care must be taken that the solution does not come in contact with the
colony. The cultures must be allowed to develop into typical colonies
before poisoning. An antiseptic formula for the purpose needs alcohol to
insure penetration of the plug, a poison to destroy the mites, glycerine to
prevent the crystallization of the poison as the alcohol evaporates, and the
dye to insure the destruction of the cotton plugs when removed from the
tubes. In our own experience we have consistently made it a practice to
wipe off the outside of all culture tubes with the above, or some other
sterilizing solution, and to poison all plugs before cultures are replaced in or
added to the collection of stock cultures.

Molds: Under humid laboratory conditions, cotton plugs, especially if
made from the absorbent type of cotton, absorb moisture. Careless han-
dling in preparation and care of such plugs often adds enough nutrients to
support growth. Steam sterilization tends to distribute nutrients. Dirt,
bacteria, and molds fall from the air upon the exposed portion of the plug.
Handling detaches spores from the colony within so that both ends of the
plug are commonly well seeded. Spore germination, therefore, may begin
at either or both ends. Outside molds may grow through and drop into the

62 A MANUAL OF THE ASPERGILLI

culture, or the culture itself may grow out through the plug and contaminate
other cultures or experiments. Surface sterilization of the outside of the
tubes and poisoning the plugs takes care of molds, as well as mites.

Spraying: Oily sprays, as selected fractions from petroleum, avail-
able from commercial sources, even kerosene, distributed with a "gun"
that produces a mist penetrating and filling all cracks, crevices, open spaces
among apparatus or furniture and clouding the whole atmosphere of the
laboratory, have been found effective in carrying down mold spores and
bacteria from the air and ridding the laboratory of mites, insects, and
vermin.

PRESERVATION OF DRIED SPECIMENS

Mold cultures lose many of their characteristic and diagnostic features
upon being dried. Nevertheless, dried herbarium specimens serve a useful
purpose in preserving type material which might otherwise be lost. Details
of morphology are often difficult to establish from such material, but group
characteristics are preserved and over-all colony appearances can be recog-
nized after many years. The retention of culture tubes or petri dishes
containing such dried specimens constitutes a reasonably satisfactory means
of preservation, and the material contained therein approximates as nearly
as is possible the cultural picture of the growing colony. Glass tubes and
dishes, however, are cumbersome and easily broken, hence may prove
unsatisfactory if frequent handling is necessary. For many years we have
employed an alternate technique with generally satisfactory results. Rep-
resentative portions of colonies grown in petri dishes are cut with a large
cork borer, lifted out with a spatula, and dropped into paper pill boxes
where they are allowed to dry. These can then be stored in larger boxes
or attached to herbarium sheets for filing. The boxes should be provided
with tight-fitting lids, and for greatest convenience should measure approxi-
mately l\ inch in diameter. Aspergilli stored in this way prove useful in
many comparative studies. They cannot, however, under any condition,
take the place of carefully handled living cultures.

Chapter VI
VARIATION

The Aspergilli are a variable and mutable group of fungi. They are
characterized by great diversity and variability as they are isolated from
nature, and an increasing amount of evidence shows that they can be made
to vary, or mutate, in the laboratory by subjecting them to a number of
different imposed stimuli. Frequently the same types of mutants or vari-
ants ultimately result under both natural and artificial conditions. Never-
theless, it is believed desirable to consider somewhat separately variations
and mutations resulting from natural causes and those resulting from
imposed stimuli.

Definition of Terms

Before entering upon a discussion of variation, either natural or induced,
it is important to define certain terminology which is to be employed. It
is recognized that our definitions will not agree in all cases with those of
earlier workers, nor do we expect that all subsequent investigators will
accept those which we propose. If the meaning in the present discussion
is clear, our purpose will have been served.

(1) The term mutant, or mutation, is used to designate a strain whose
source is actually known and can be verified. Furthermore, it is limited to
those substrains which originated as sharp breaks from parent cultures
(usually interpreted as gene mutations), and in successive culture genera-
tions retain their distinguishing characteristics unaltered. This may or
may not have a taxonomic connotation. Upon occasion it is used in essen-
tially the same sense as "variety". To illustrate, A. nidulans mut. albus,
A. fumigatus mut. helvola, A. niger mut. cinnamomeus, etc., are used as
Latin names to designate forms which differ from the parent species in
certain striking details. In other instances it is used to identify a type of
change, rather than to designate a particular and isolated strain resulting
from such change. It is considered correct to refer to artificially produced
albino, yellow, and buff-colored strains of A. terreus as mutations (see p.
75), since they are constant in character and are known to have originated
from a cinnamon-colored parent culture, wholly representative of the
species; and we believe it represents good judgment to refrain from assign-
ing Latin designations to each of them. The term mutation, then, refers
to altered strains of constant character and known lineage, whether or not
the y are given Latin designations.

63

64 A MANUAL OF THE ASPERGILLI

(2) The term variant, or variation, is used loosely and reference to it in
this manual is not, in all cases, entirely consistent. In general, however, it
is applied to subcultures, or strains, arising through gradual change from
well-defined strains of identifiable species. The characters of a variant,
then, are not generally stable but subject to continued change and further
variation. As used by us, the term has no taxonomic implication and can
be considered essentially synonymous with the term "saltant" which is so
commonly employed in reports on variation in the Fungi Imperfecti.
Variants frequently appear as colony sectors, overgrowths, or other local-
ized areas of changed appearance or texture. When isolated in pure cul-
ture, they may or may not retain their distinguishing characteristics.

NATURAL VARIATION

Cosmopolitan species and groups of Aspergilli show adaptability to wide
ranges of environmental conditions. As these molds are isolated from
nature, variation among the members of any species, series, or group is
regularly encountered. Such variations commonly differ in degree rather
than in basic characters, and one can distinguish a series of intergrading or
bridging forms. Even striking isolates are often unmistakably allied with
some well-defined species or group in this manner. Such different but
intergrading forms arising in nature can be considered as natural variants.
Natural variants of a similar kind can frequently be obtained in laboratory
culture by selective isolation and cultivation from sectors or other areas of
atypical growth, or by single spore isolations. Distinction must be drawn
between differences in appearance, morphology, and habit of growth result-
ing from inherent differences between strains, and alteration in colony
character in response to changes in the composition of the culture medium
or other environmental factors. Rigid comparative culture is often neces-
sary to distinguish between the two. For the present discussion, we are
concerned with differences that are more fundamental than direct tempo-
rary responses to artificial stimuli (ecads); but the latter, unless carefully
evaluated, may appear no less real. Rightly or wrongly, Blochwitz (1930,
p. 247) comments that .4. flavus,1 in specimens collected in the Botanical
Garden at Buitenzorg, was called A. penicillopsis (Henn.) Rac; in the
Botanical Garden at Singapore, *S. vitellina Ridley; in India, S. corolligena
Massee; and in Columbia, .4. delacroixii Saccardo.

Occasionally isolations are made from laboratory cultures which represent
sharp "breaks" from the parent strain, and since they are constant in subse-
quent culture, they may be considered as true mutations. Representatives

1 The name of the original describer is only used for specimens or strains in culture
which were definitely attributed to the describer. A. flavus, A. niger, A. terreus, etc.,
are series concepts as used here.

VARIATION 65

of such natural mutations which originated in the absence of any artificially
imposed stimuli are YuilPs A. fumigatus var. helvola (1939), A. nidulans
var. albus (1939), and Cladosarum olivaceum (1938).

Intra-strain Variation

A certain amount of variation can be expected to occur in any given strain
of Aspergillus. In certain species and strains this is very limited and cul-
tures can be re-cultivated repeatedly upon a variety of media, and at
different temperatures and H-ion concentrations without evidences of visi-
ble change other than those resulting from the immediate effects of the
altered environment. Many strains of Aspergillus niger are characterized
by such comparative stability. Other species and strains are subject to
continual variation with differences in character and rate of growth appear-
ing rather abruptly as sectors or overgrowths, or gradually developing as a
progressive alteration in the general aspect of the whole culture. By suc-
cessive and selective subculturing, strains of A. alliaceus and A. ochraceus
showing a marked difference in sclerotium production can be obtained. In
like manner, strains of A. itaconicus Kinoshita can be secured which are
almost completely sterile upon all media tested. The same is true of A.
granulosus Raper and Thorn, A. flavipes, etc.

Working with a strain of the ascosporic species, A.fischeri, Greene (1933)
isolated 448 single spore cultures and among these found variant progeny
of two main types: (1) cultures producing very large, scattered perithecia
as opposed to the typical picture of many small perithecia, and (2) cultures
producing conidial structures in profusion, but forming few perithecia and
these tardily. The second type was fairly stable, whether derived from
single ascospores or conidia. The first type was variable, in some cases
reproducing the characters of the variant parent, in others reverting to the
character of the original stock culture.

Hansen (1938) and Hansen and Smith (1932) have studied many of the
Fungi Imperfect! rather exhaustively and report that single strains of these
fungi are basically composed of a mycelial (M) type and a conidial (C) type.
By proper techniques the two forms can be separated and recombined at
will. While it has not been explored as yet, the possibility exists that the
same condition may prevail to a limited degree in the Aspergilli, and that
this may account for a certain amount of the intra-strain variation en-
countered.

Back of variability in molds, many lines of discussion have been devel-
oped. Buller (1933) has frequently called attention to the unmeasured
possibilities of nuclear and cytoplasmic disturbance from the commonly
observed phenomenon of anastomosis. Vegetative hyphae belonging to the
same mycelium (mycelium derived from a single spore), or different mycelia,

66 A MANUAL OF THE ASPERGILLI

throw out branches which fuse without showing any other sign which might
suggest a sexual process either before or after fusion. Anastomosis is
usually observable, if at all, in the rapidly growing area where many spores
placed as an inoculum are developing into one colony. By transferring
large numbers of spores from an old culture to a new one, most of the
Aspergilli studied by us have shown fairly consistent repetition of colony
characters and conidial morphology and have been maintained for a long
time with little or no observable change. Other organisms handled in the
same manner have not been successfully maintained with the morphology
originally studied. Differences in behavior can be attributed to variability
between strains.

Intra-species Variation

Variation within the species is very prevalent and is well marked in many
cases. When a large number of isolates of any particular species or series
is collected, one can regularly expect to find among them wide variations in
color, amount of sporulation, and in their general habit of growth. Usually,
however, such variation is graduated, and strains representing various
intermediate steps between the extremes are to be expected. While it is
by no means unique, we may use A. terreus as an example, since a very large
number of strains belonging to this species have been isolated and observed
in plate and tube cultures during the past two years (fig. 16). Colonies of
the type strain, and of the great majority of isolations made from nature,
are plane, cinnamon in color, very heavy sporing, with conidial heads arising
directly from the substratum in an even and close stand. An occasional
isolate is much brighter in color, approximating xanthine orange (Ridgway,
PL III), but in all other respects it is fairly typical. It is believed to repre-
sent a form such as that described by Blochwitz (1934) as A. boedijni, and
in the present manual we have designated it as A . terreus var. boedijni. In
each of the above cases, the production of abundant fruiting structures
follows closely the advancing margin of the growing colony and there is little
or no continued growth of mycelium except in the marginal area. Certain
other strains are quite floccose; conidial heads are typical in form and color
but are greatly reduced in number and are borne upon aerial hyphae. Shih
(1936) was probably working with such a culture when he described the
variety A. terreus var. floccosus. We feel that the forms are sufficiently
distinct to warrant maintaining his variety. Still other strains possess
abundant but fairly close-, rather than loose-textured mycelia and bear
abundant but very pale buff-colored conidial heads. The vegetative my-
celium in this form is bright yellow and for this reason we have designated it
as A . terreus var. aureus n. var. (see p. 198) . If one should examine only the
type strain and these three atypical forms, it is entirely probable that one

VARIATION

67

would describe them as four distinct species. Actually, however, they are
not sufficiently distinct to warrant specific rank, for they represent only
extremes of variations along three divergent lines with numerous inter-

Fig. 16. Intraspecies variation in Aspergillus terreus. A, A. terreus, typical
strain NRRL No. 265, characterized by heavy conidium production and colonies
cinnamon in color. B, A. terrreus var. boedijni, NRRL No. 680, characterized by
deeper colonies and conidial heads near xanthine yellow. C .A . terreus var. floccosus,
NRRL No. 1921, characterized by loose floccose colonies and spore heads light
pinkish-cinnamon in color. D, A. terreus var. aureus, NRRL No. 1923, character-
ized by yellow floccose colonies and comparatively few cream to buff-colored conidial
heads.

grading strains aligning them, almost without interruption, with the typical
form itself.

Another series of variants in our collection shows gradation from the
usual radiate head and conidiophore of A . sydowi to the simple mono-verti-
cillate penicilli of Penicillium restrictum Abbott. The tendency to form

68 A MANUAL OF THE ASPERGILLI

reduced conidial apparatus is observed in all of the strains of A. sydowi
examined. Over a period of many years variants have exhibited all
gradations in colony appearance from typical A. sydowi to the aspect of P.
restrictum of Abbott except for the presence of an occasional conidiophore
and head of .4. sydowi. Repeated cultural tests exclude contamination.
These variants occur in nature, they produce abundant conidia, and they
undoubtedly maintain themselves successfully in the field.

Aspergillus fumigatus presents a somewhat similar condition. In this
species, typical cultures produce heavily sporing, velvety colonies that are
dark green in color and show almost no aerial mycelium. Other strains
commonly isolated from nature produce very floccose colonies and bear
comparatively few conidial heads (fig. 37). These heads, however, are
typical in form and in dimension. Strains possessing this contrasting
character are relatively stable in culture, but in this species, as in A. terreus,
all degrees of intergradation are found between this floccose type and strains
entirely typical of the species. The same story is repeated in other species.

Appreciable variation can normally be expected among the isolates of
any of the very abundant and cosmopolitan species. Such variant strains,
however, are the exception rather than the rule, since the great majority of
isolates are quite typical of the species. Attention was called to this fact
in our study of the A. glaucus group (1941).

When grown in comparative culture, strains successively isolated as repre-
senting a particular species usually show enough difference to give each
strain a kind of individuality. Exact identity, point by point, is not ex-
pected. Such strain variation may be incidental and unimportant, or it
may be correlated with activities which make one strain a valuable agent
in an industrial process and the other worthless.

Intra-group Variation

Inside the different groups of Aspergilli one normally finds somewhat
similar but wider variations than those seen among the strains constituting
any particular species. To what degree species in nature have developed
by mutations and by progressive variation can only be guessed. We do
know, however, that the species within a group, like the strains or varieties
within the species, are regularly bridged by intermediate forms which render
it difficult to establish sharp and immutable lines of separation. One can
almost cite it as a rule that the definiteness with which one regards a species
is inversely proportional to the number of strains of that species which have
been examined. Still species are necessary as guide-posts — as fixed points
around which closely related organisms showing a certain but limited
amount of variation can be grouped.

The Aspergillus flavus-oryzae group can be taken as illustrative of the
type of variation to be expected within a group of the Aspergilli. Thorn's

VARIATION 69

culture No. 113 (XRRL No. 447) of A. oryzae, received from Baarn and
believed to stem from Cohn's original strain, is a very floccose, loose-textured
culture bearing comparatively few, small, light yellow to tan heads. Co-
nidiophores are long, ranging up to 2 to 5 mm., and are very thin- walled.
There is only a trace of green even in individual heads, and in general aspect,
the culture normally shows no green color. Aspergillus flavus in its typical
form is not floccose and is very heavy sporing. Conidiophores arise directly
from the substratum in a close stand, are usually 1 mm. or less in length,
and are comparatively heavy walled. Colonies are regularly in yellow-
green shades and range from light to comparatively dark green (see species
description). A. parasiticus Speare goes even farther. Colonies are very
dark green in color. Conidiophores usually range from 200 to 400m in
length, and sterigmata are typically in a single series, whereas they may be
in a single or double series in A . oryzae and A . flavus. In the opposite direc-
tion, but markedly different from A. oryzae is A. effusus Tiraboschi. Typi-
cally this is very floccose and comparatively light sporing, with heads borne
upon short conidiophores which arise from the loose aerial mycelium rather
than from submerged mycelium in the substratum.

The cultural pictures of these species are fairly characteristic. Yet, it is
practically impossible to take a large collection of 100 or more strains and
separate them into these species with any degree of confidence or satisfac-
tion. The difference between A. oryzae and A. flavus is bridged completely
by a series of intermediate forms showing all degrees of variation between
the two strains selected as typical. Xomenclature in this group is then
further complicated by the fact that among the great collections of these
forms obtained from the Orient, and designated A. oryzae, the majority of
forms are somewhat intermediate between A. oryzae and A. flavus as de-
picted above. A similar series of intermediate forms bridges completely
the gap between A. flavus and A. parasiticus. There is no sharp line of
demarcation between any of these species, still they are not one and the
same, and to attempt to lump these diverse forms together into one species,
as Xeill (1939) has done for the ^4. glaucus group, intensifies rather than
reduces the difficulties encountered.

Intra-group variation is also particularly marked in the .4. niger group.
During an extended period of study and observation of molds in culture,
Biourge, who was a discriminating collector, accumulated 63 strains of
black Aspergilli which suggested sufficient individuality to be deemed
worthy of further study. These were turned over to Mosseray when he
entered Biourge 's laboratory. He assumed that he had before him all of
the black Aspergilli possible to collect and, knowing that Biourge had
selected each of them because it seemed to have some special character, he
undertook a taxonomic study to define those characters and to organize
them into a systematic presentation (1934a). His paper lists 35 species of

70 A MANUAL OF THE ASPERGILLI

which 25 were either described as new species or new combinations. His
findings in A. niger are fairly illustrative of the same type of study in other
groups (compare Thorn and Currie [1916] for A. niger; Thorn and Church
[1921] for A. flams.

Mosseray based his primary separations upon conidial sizes, shapes, and
markings, while secondary and tertiary separations were based upon co-
nidiophore lengths and the characters of colonies in tube cultures on
Biourge's "Raulin-neutre gelose" — a variation of the classic Raulin solution.
Biourge and Simonart demonstrated the entire series to one of us (C. T.)
showing how wrinkling and granulation of mycelium, intensity and changes
of secreted color, shades of color in the conidial area, lengths and propor-
tions of conidiophores and heads, and their distribution over the mycelium
gave to each strain an individuality which had been repeated in successive
cultures over a considerable time. We raised just one question, "What
would you do with the next thousand?"

The large majority of all isolations of black Aspergilli conform within a
range of minor variation with the general van Tieghem concept, — i.e.,
black-brown colonies with conidiophores and heads giving the general
structure and measurement of parts found in the classical description.
Then, in contrast with these, there are shades of colony color from the coal
black of A . carbonarius through shades of purplish-black to the brown of A .
ferrugineus Fuckel or to the lighter shades of Schiemann's mutants (pp.
223-224) . Some of them produce no colors in the substratum and reverse
of the colony; some show yellow in traces; others are persistently deep
orange, giving the whole a yellowish appearance. Or, again, the agar and
mycelium may develop a red-brown or "mauve" shade of violet.

Conidiophores in the usual type of culture reach nearly enough the same
length to give the effect of a field of grain. But their length may be quite
short and the heads seem to be borne directly on the substratum, or they may
be several millimeters in length with the heads borne well above the sub-
stratum and correspondingly large. Between these extremes, every varia-
tion can be found. Conidiophores may be scattered thinly over the
vegetative mycelium, collected in a zone at the border, or crowded in
the center.

The vegetative mycelium may grow as a flat felt (plane) or may be vari-
ously wrinkled, sulcate, or buckled. In a smooth or plane colony the
mycelial cells seem to stop growing early — the colony extends only at the
margin. In the plane colony intercalary growth (i.e., the formation of new
cells in the filament, or the lengthening of the old cells), and the production
of new branching ceases. Such mycelia ordinarily produce one crop of
conidial heads, beginning at the center of the colony and progressively
developing toward the margin until the medium is exhausted or some
inhibiting factor paralyzes growth. Marginal growth in such a colony

VARIATION 71

often shows longer and fewer conidiophores and larger heads than in the
central area.

Within the group with the usual structures still recognizable, many
variants with contrasting features appear. Heads with very long primary
sterigmata, which are sometimes septate, appear in A. carbonarius (Bainier)
Thom, A. pulchella Speggazini, or A. tubingensis Mosseray. The primary
sterigmata may grow out into sterile filaments as in Mosseray 's figure for
A.ficuum (Reich.) Henn.; much more commonly, some primary sterigmata
grow out as tiny conidiophores and produce little heads, often consisting
only of a cluster of simple sterigmata and conidial chains. Thus, sterig-
matic changes may run from the simple sterigmata of A. japonicus Saito,
A. luchuensis Inui, or A. malvaceus Mosseray, where only some are double,
to other species showing the widest range in length and arrangement.

Another group of variants show marked suppression of the ordinary
structures expected. Strains in which conidiophore formation has been
reduced or almost suppressed have been studied in continuous culture.
Such colonies showed an occasional long conidiophore and large head, con-
forming to the A. niger pattern, produced at the end of the colony growth
period. Meanwhile the mycelium was fully covered with irregularly
branching hyphal elements bearing single sterigmata variously placed,
groups of sterigmata, or penicilloid clusters of sterigmata each bearing a
short chain of conidia showing the characteristic markings of the group.
Transfers from the simplest form developed the complex or A. niger ele-
ments. Transfers from the large heads brought a recurrence of the re-
duced type of fruiting. No method of selection tried brought back the
typical A. niger aspect, and cultures of this type appear to represent de-
generate forms.

It would appear, then, that a general type or morphological picture when
found dominant in large numbers of natural isolates can be regarded as
typical for a species of Aspergillus. It is recognized that marked diver-
gences from such types occur under the unrecorded stimuli of nature.
Some of these forms succeed in establishing themselves as permanent
elements of the microflora and thus become successful as species, or va-
rieties. Others do not digress quite so markedly and thus constitute
intermediate or bridging forms. At the same time marked changes can
be induced by the application of artificial stimuli. Where the origin of
such altered strains is known, they are commonly regarded as mutants.
Were they isolated directly from nature and their previous history not
known, it is probable that they would be considered as separate varieties
or even species.

Natural Mutation

While most of the mutants which we recognize as such have originated
in the laboratory as the result of certain artificially imposed stimuli (or

72

A MANUAL OF THE ASPERGILLI

drastically altered conditions of growth), and hence can properly be termed
induced mutations, a number of well authenticated cases of natural muta-
tion are known. In 1939 Edward Yuill described as .4. fumigatus var.
helvola a buff-colored mutant of this species isolated by him in 1937 (fig.

Fig. 17. Natural mutations. A, Portion of a colony of Aspergillus niger in which
a tan spored mutation appeared as a V-sector : Ai, typical black head of parent strain;
A2, tan head of naturally occurring mutation; X 25 approximately. B and C , Typical
strain of Aspergillus fumigatus and a naturally occurring mutation discovered and
described by Edward Yuill as Aspergillus fumigatus mut. helvola. The mutations
in both species have proved completely stable in continued culture.

17 C). In the same report a white-spored mutant of A. nidulans, isolated
in 1937, was described as A. nidulans mut. alba. In both cases the mutants
developed as natural phenomena without the application of any artificial
stimuli — in the former case as a single head, in the later case as a group
of heads, and in both cases from wholly typical strains (fig. 17).

Plate II

-Mutations in Aspergillus terreus Thorn produced by irradiating conidia with ultraviolet light. Colonies
tin >\\ n upon C'zapek's solution agar at room temperature for two weeks. A (upper left), Aspergillus terreus,
XRRL No. 265, unirradiated stock culture. B (upper right ), Albino mutant, producing white conidial heads
but retaining the basic cultural and morphological characteristics of the parent culture. C (center left), floc-
cose, yellow-white mutant producing few and atypical conidial heads. D (center right), Leathery mutant,
producing tough, close-textured colonies and very few and atypical conidial heads. E (lower left), Nitrate
mutant, a form unable to use nitrate nitrogen, producing thin spreading colonies and few but entirely typical
heads. F (lower right), Thiamin mutant, a form unable to produce thiamin, producing thin, spreading
colonies and very few and atypical conidial heads, see text p. 75. (Color photographs by Haines, Northern
Regional Research Laboratory. Reproduced through co-operation of Chas. Pfizer & Co., Inc.)

VARIATION 73

In studying a group of cultures three years ago, the authors noted a few
tan heads in the form of a Y-sector in an otherwise typical black colony of
A. niger (fig. 17A). Isolations from a tan head reproduced the mutant head
characters, and repeated transfers of this strain have proved consistently
stable over a period of three years. The mutant strain cannot be
distinguished from A. niger mut. cinnamomeus (A. cinnamomeus of Schie-
mann).

In dealing with natural as well as induced mutations, we have endeavored
to limit the use of the term mutant to forms whose origin was definitely
known. Blochwitz (1934, 1935) was not so precise. Under the name
A. glaucus mut. alba he refers to a white-spored member of the A. glaucus
group. He believed Aspergillus giganteus Wehmer to represent essentially
a long-stalked .4. clavatus, hence designated it A. clavatus mut. giganteus.
This treatment may be justifiable. We believe, however, that until their
origin from other and well-marked species can be proved, it is wise to con-
tinue to recognize as species these very distinct forms that are isolated from
and are able to maintain themselves in nature.

Cladosarum: The most striking variant, or mutant, ever described in the
Aspergilli is Cladosarum olivaceum of Yuill and Yuill (1938) . This appeared
in a culture of A. niger growing on bread at 28° C. (Personal correspond-
ence). Its colony, conidiophores, vesicles, and primary sterigmata are
those of Aspergillus. The secondary sterigmata, instead of producing co-
nidia, thrust out cells which are essentially the same in morphology as the
secondary sterigmata themselves; the same procedure is then repeated
several times. Occasionally, however, a terminal cell changes and thrusts
out several equal cells; in other words, it resumes the function of a primary
sterigma. The new secondaries repeat the process of producing chains of
cells each resembling the basal cell with the aspect of a sterigma not a conid-
ium, and always with the youngest cell at the tip of the chain. In Asper-
gillus the sterigma which produces a chain of conidia always produces the
new conidium at the base of the chain, shoving the next most recent farther
out. Differing then from Yuill 's interpretation, Cladosarum produces no
conidia, however readily any cell detached from the mass may grow.

In Aspergillus the ordinary nuclear procedure in conidium formation
involves mitosis in the sterigma actually producing the conidia. After
each mitosis one daughter nucleus migrates through the tube into the new
spore in which it "rests" until that spore begins to germinate. The other
nucleus remains in the sterigma and repeats the process. This goes on
until there may be a chain of 200 conidia — the oldest at the outer end, the
newest directly attached to the sterigma.

In the absence of cytological study, one may offer the following
hypothesis. In "Cladosarum" the nuclear procedure must be reversed.

74 A MANUAL OF THE ASPERGILLI

The same mitosis occurs. One active and one resting nucleus result, but
the resting nucleus remains in the sterigma while the active nucleus moves
into the newly forming cell. This determines the course of development.
The active, multiplying nucleus is always in the newest cell formed.

Previously Barnes (1928) had stimulated a strain of the A. glaucus group
(identified by us as A. amstelodami, 1941) by heat, and reported certain
mutants which were deposited with Dr. Westerdijk at Baarn. Among
them, under the designation "Creamy" (NRRL No. 143), a mold with the
morphology of Cladosarum appeared. It was obviously derived from
some A. glaucus strain and is, in so far as the writers are aware, the only
other appearance of the Cladosarum structure ever discovered. Barnes
does not appear to have recognized its contrasting structure.

Since no collector has reported this type of mutant in nature, it must
either be very rare or be unable to maintain itself in a competitive environ-
ment. However readily such mutants may be maintained in the labora-
tory, they would rarely reach the second generation in nature on account
of lack of spores. The name Cladosarum was thus applied to a defective
organism (zoologically designated a "monster"), which does not become a
component of any natural flora, hence taxonomically the name should
be untenable.

INDUCED VARIATION

Striking mutations have been obtained from various species of the Asper-
gilli by subjecting them to artificially imposed stimuli. Schiemann (1912)
was among the first to draw attention to the possibilities inherent in this
approach. By subjecting a strain of A. niger to various concentrations of
potassium bichromate, she was able to produce two striking mutations
which she designated according to color, A. fuscus (= A. niger mut. schie-
manni of this manual) and A. cinnamomeus (= A. niger mut. cinnamomeus
ibid.), respectively. Both cultures have remained stable in our hands, and
in various collections, throughout the 32 years since their original isolation.
A third "mutation" designated A. niger var. altipes could not be
distinguished from other strains of black Aspergilli isolated from nature —
the parent strain was not seen. Working with a member of the A. glaucus
group designated Eurotium herbariorum Wigg., Barnes in 1928 reported
the production of a series of variations by exposing spores to heat. While
there are reasons for questioning the correctness of some of Barnes' inter-
pretations and conclusions (see Thorn and Raper, 1941), there is evidence
that he succeeded in producing a mutant which, in its habit of growth and
in the character of the fruiting structures developed, bears a striking re-
semblance to a form subsequently isolated from A. niger by the Yuills
(1938), and described by them as Cladosarum olivaceum, genus and species

VARIATION 75

new. Galloway (1933) obtained marked variation in colonies of Asper-
gillus terreus by growing them upon media containing flour to which was
added 0.003 to 0.005 per cent of salicylanilide.

Thorn and Steinberg (1939), and Steinberg and Thorn (1940a, 1940b)
in a series of experiments, applied chemical stimulants to a strain of A.
niger (Thorn No. 4247: NRRL No. 334) which had been in the collection
many years without noticeable change. From the cultures resulting,
Steinberg picked out and purified for study all variants he could observe
with the naked eye and with the aid of a handlens. In examining a
fruiting area of a colony, general changes were not common; ordinarily an
occasional head changed color, long or short conidiophores appeared in
spots, gross malformation showed as areas of no fruit, or too much fruit,
or color effects in the mycelium. These were picked out and grown in
successive cultures. Expressed in terms of morphology, the most striking
feature of the tested culture was disturbance of uniformity. The same
types of changed aspect were reproduced many times. In general, they
followed the same lines as have been described as present in the natural
series selected by Biourge or in the collections of Mosseray at Brussels.
In general, these changes were destructive in character and included large
numbers of "injury mutants", or variants, which reverted in subsequent
transfer to the original aspect of Steinberg's culture. Sodium nitrite was
the most effective agent used. Some isolations, however, represented clean-
cut mutations. Strains of A. fumigatus with albino heads were obtained.
Forms of A. niger with light brown to cinnamon-colored spore heads, es-
sentially like those earlier obtained by Schiemann (1912), those obtained
by Whelden (1940), and those subsequently obtained by Raper, Coghill,
and Hollaender (see below) from members of the same group, were likewise
isolated. These have remained stable in culture for the four years that
they have been in our collection.

Whelden (1940) succeeded in obtaining a series of mutants in A. niger
by bombarding conidia with low voltage cathode rays and subsequently
isolating colonies which developed from such irradiated cells. Forms pos-
sessing heads in various brown shades, rather than black, were isolated,
as well as one giant form with conidial structures appreciably larger than
the parent. By means of ultra-violet irradiation of spores, Raper, Coghill,
and Hollaender (in press) obtained mutants which produced tan-colored
conidial heads but otherwise closely resembled the parent strain. In a
more exhaustive study of A. terreus (PI. II, A), the same investigators suc-
ceeded in isolating a number of striking and markedly different mutations.
These included albino forms with colorless conidial heads (PI. II, B); forms
with pale buff-colored heads; yellow, yellow-white, floccose forms with few
and smaller conidial heads (PI. II, C); forms producing very thin, sparsely

76

A MANUAL OF THE ASPERGILLI

sporing colonies; forms producing restricted colonies characterized by the
production of an excessive amount of orange-brown exudate; and forms
with leathery, close-textured colonies bearing very few conidial heads
(PI. II, D). Alterations in microscopic details commonly accompanied
these changes in colony appearance (fig. 19). In addition to these morpho-
logical mutants, which were found to be stable when checked through ten
successive transfers over a period of 12 months, various physiological
mutants were also isolated. These included forms unable to utilize nitrate
nitrogen (PI. II, E) but able to grow and sporulate normally upon media
containing ammonia nitrogen, and a form unable to synthesize thiamin
(PI. II, F). Upon Czapek's solution agar containing sodium nitrate and
sucrose, each of these is strikingly different from the parent strain; upon

A

Fig. 18. Induced mutation. A and B, Aspergillus niger group, strain NRRL No.
67: A, parent culture growing on Czapek's solution agar, 10 clays, room temperature;
B, tan-spored mutation of same produced by ultraviolet radiation.

malt extract agar, which contains adequate amino nitrogen and thiamin,
neither could be differentiated from the parent (Raper, Coghill, and Hol-
laender, in press). Thus the need for comparative study and examination
upon a variety of media is apparent, while the importance of such studies
in reliable taxonomic work cannot be over-emphasized. In cultures of
Aspergillus terreus resulting from irradiated spores the capacity to produce
itaconic acid varied from zero in some isolations to levels somewhat above
the parent culture in others. The majority of such isolations produced
yields somewhat lower than the parent strain, many produced yields ap-
proximately equal to it, while a very few produced superior yields (Lock-
wood, Raper, Moyer, and Coghill, in press).

Based upon our own investigations and the published reports of other

VARIATION

77

workers, certain observations of a summary character can be made regard-
ing variation in the Aspergilli:

1 . Aspergilli include strains and species adapted to a very wide range of
environmental conditions. Such conditions may influence materially the
cultural and morphological characteristics of these molds.

ft '

B

,A

c

Fig. 19. Photomicrographs showing details of structure in the conidial heads of
the parent strain, and in two selected mutations of Aspergillus lerreus (NRRL No.
265) produced by ultra-violet, radiation, X 600 A, Typical heads of non-irradiated
parent strain. B, Mutation in which conidium formation is incomplete and cells
adhere in long chains in liquid mounts. C, Mutation in which many fruiting struc-
tures develop vesicles but often fail to produce sterigmata and spores.

2. Under natural conditions, great numbers of variants appear along with
occasional sharply separable forms, or mutants, which are definitely of
species rank. The extent to which natural mutations may account for
described species is a matter of conjecture.

3. The Aspergilli can be made to mutate in the laboratory by subjecting
them to a variety of different excitants, or stimuli. Induced mutations may
parallel some of those found in nature and described as species.

78 A MANUAL OF THE ASPERGILLI

4. Particular species of the Aspergilli tend to mutate along certain
definite lines, e.g., the production in A. niger of forms with tan to light
brown spore heads (Schiemann, 1912; Steinberg and Thorn, 1940; Whelden,
1940; and Raper, Coghill, and Hollaender, in press), and the production
mA.fumigatus of forms with colorless spore heads (Yuill, 1939; and Stein-
berg and Thorn, 1940a). The type of mutant produced is not governed
by the type of treatment given, although the number of mutations produced
is strongly influenced by this factor.

5. Great variability in biochemical activity is encountered among strains
isolated from nature, but these differences are rarely linked with specific
morphological changes.

6. The Aspergilli can be made to mutate physiologically as well
as morphologically by the application of various stimuli. Physiological
mutations may conceivably be of tremendous importance in the develop-
ment of improved strains for fermentation processes.

7. Despite natural variation, most strains of Aspergillus when subjected
to critical transfer and maintenance under rigorous culture conditions can
be kept for many years with constant colony appearance, stable
morphology, and dependable biochemical activity.

PART II

THE MANUAL PROPER

Chapter VII
THE USE OF THE MANUAL

Since the purpose of this manual is to facilitate the identification of
Aspergilli as they are isolated from nature and as they are encountered in
the investigation of special problems, the procedures and considerations
involved in the use of the manual must be discussed. The general mor-
phology and structural details found in the spore-producing apparatus of
the Aspergilli have been described and figured in Chapter III. Com-
plexities in the specific combinations of these characters found in the exam-
ination of moldy material and in the isolated colonies of individual strains
makes desirable a summary outline of the exact observations to be made
in describing an Aspergillus. Such a descriptive sheet is presented as page
82. For practical use, a standard sheet of record paper is folded over on
the left-hand margin for about 5 cm. The column of observations desired
is written upon this marginal fold ; a fresh sheet of paper is slipped under the
fold and the descriptive data are filled in, appearing exactly in the same
order for each strain studied. A single glance at the sheet shows the dis-
crepancies, if any, in the descriptive data obtained. With such a sheet
properly filled out, the keys to groups and within groups facilitate the
placement of the strain in its proper group first, then its allocation to species
within the group.

Such descriptive sheets, to have comparative value in species diagnoses
must present their data in standardized terms. It has, therefore, been
necessary to define and illustrate the morphological terms accepted in this
manual, and to indicate as synonyms in the chapters on morphology the
usages of various describers of Aspergilli back over the 200 years since
Micheli.

Identification from specimens : The field mycologist working with speci-
mens collected and examined fresh or dried will often find completion of a
technical description very difficult and some observations impossible. One
with long acquaintance with the Aspergilli may place his specimen to the
group or aggregate species correctly, but even such workers are frequently
puzzled. If the organism is deemed important, the fresh or recently col-
lected specimen should be taken to the culture laboratory to insure its
isolation and preservation in pure form. The descriptive data at hand
should then be checked and supplemented from the pure culture.

To identify an unknown Aspergillus, the worker needs pertinent data
which will permit him to interpret his mold in terms of species already de-

81

82

A MANUAL OF THE ASPERGILLI

scribed— including the observations essential in a species characterization.
For convenience such data may be indicated vertically upon a descriptive
sheet which is elaborate enough to include observations that are regarded
as useful. Measurements should be presented as ranges encountered in
the examination of many units, not as exact and single measurements of
individual cells or structures.

Primary sterigmata

measurements

arrangement

color
Secondary sterigmata

measurements
Conidia

color

measurements

markings
Perithecia

color

size

shape
Ascospore

color

size

markings
Sclerotia

color

size

Aspergillus — identifying number or marks.
Culture medium or natural substratum
Temperature of incubation
Colony characters
Rate of growth
Texture
Mycelium

submerged
floccose
color: above
reverse
Heads
color
form

measurements
Conidiophore
length
diameter
wall: thickness
markings
color
Vesicle
shape
size
color

With such a descriptive sheet before him, the user of the manual finds
that the Aspergilli have been arranged into a series of natural groups (fig.
20), each containing one to several species aggregates. Each group in-
cludes species with varieties, and at times mutants, having a series of es-
sential characters in common. These groups have been arranged as nearly
as possible in natural order, based upon the presence or absence of certain
contrasting intergroup characters.

These major separating characters are usually evident and positive.
Nevertheless, individual species are found in which certain of these charac-
ters are reduced to vestigial or apparently suppressed, yet which show so
many characters allying them with a particular group that such placement
is more logical than any other. Such species must sometimes be arbitrarily
placed, and their possible affiliation with other groups indicated both in the
discussion of the species and in the discussion of the related group.

THE USE OF THE MANUAL 83

Species

The species concept in Aspergillus is very difficult to define in tangible
terras. In this manual, the species names already in use have been pre-
served wherever possible. The actual material originally described under
a particular species name (i.e., type material) exists for but a few species.
If such material exists, it is more important as fixing one point, one individ-
ual strain in a series of intimately related variants, than tying the name
to extremely definite morphology. If such material is not known, compari-
son of large numbers of strains in pure culture with authoritative descriptive
information, supplemented by laboratory usages coming down from the
original describer, usually fixes the series or form intended.

From such composite sources it is commonly possible to establish a fairly
concrete morphological aspect based upon ranges of color, differences in
structure, and variations in spore measurement which are repeated in great
numbers of isolates. In such series of isolates, there are no sharp lines of
demarcation when large numbers of strains are brought together. Within
our concept, a single strain may show much of this variability within its
colonies in culture, or it may reduce or suppress certain characteristics and
intensify others. Such variants have often been given species rank by
workers unaware of the existence of other variants completely bridging
the gap between such forms and other members of the series. Great dif-
ferences in biochemical activity may be shown by different strains with or
without contrasting morphology. Nomenclature based upon an assumed
correlation of a particular cultural aspect with industrial significance has
been offered but has proved utterly unreliable in identifying an organism
if lost, or in seeking a new strain to serve the same purpose.

Two contrasting tendencies in classification are always encountered.
In the Aspergilli these may be represented by Mosseray (1934a) who found
diagnostic marks to distinguish 35 species among 63 cultures of black
Aspergilli in the collection of Biourge at Louvain. He later received many
more variants and faced the question whether to try to describe them all or
abandon the field. He admitted inability to write descriptions explicit
enough to identify them all. In contrast, Neill (1939), disregarding asco-
spore measurements and markings, "lumped" all of the A. glaucus group
into A. glaucus Link. Likewise, all of the black Aspergilli were considered
as A. niger van Tieghem. Forms that he did not happen to recognize as
belonging to one of his groups were discarded. These are extremes.

With abundant living material before him, the student of the Aspergilli
can usually recognize as representative a reasonable number of forms which
can be described in tangible specific terms. Commonly, forms which actu-
ally play a significant role in nature or in biochemical processes can be
selected as the points around which such species descriptions are drawn.

84 A MANUAL OF THE ASPERGILLI

On the other hand, forms such as A.janus, A. itaconicus, A. lutescens, etc.,
while probably rare in nature, possess sufficiently distinctive morphology
to warrant species recognition irrespective of other considerations.

Varieties

The taxonomic term, variety, is used here to designate any homogeneous
member of a species complex which carries most of the diagnostic characters
of the species but maintains one or more clearly defined differences in
particular characters. For example, variety alba is used for certain strains
of particular species in which the characteristic color of that species is
absent.

There is little agreement in the literature in the application of the term,
variety; certain authors use the term to indicate their belief that one form
with particular morphological characters had its origin from another.
In such cases, the belief is hypothetical, not a matter of observation.
Sometimes previously known species were merely moved to varietal stand-
ing without specifying the characters upon which the decision was based.
Such changes are reduced to synonymy or, if entirely unsupported, are
occasionally ignored in this manual. The term variety is only useful if
definitely associated with a clearly defined variation in structures within an
otherwise homogeneous series of strains.

Mutations, or Mutants

The term mutation, or mutant, is only recognized here for forms resulting
from a sharp break in morphology (including color) from known structures
characteristic of a species, to a definitely altered and inherited contrasting
structure. Obviously the only excuse for the term in taxonomic usage is to
designate the origin of the form studied. If the source of such a variant
were unknown, the taxonomist would designate the form present as a
variety or species, depending upon the nature and importance of the changes
encountered. The increasing number of studies in experimental evolution
make recognition of induced variation taxonomically necessary.

New Species

The discriminating collector will occasionally find an organism markedly
divergent in characters from any described form. Usually these diver-
gences leave the organism readily recognized as a member of one of the great
groups. If the differences in aspect and detail of structure separate such
a form from the other described members of the group, and if the form is
found often enough to prove that it has a place in nature, description as a
new species is warranted. Similarly, an occasional form, either by sup-

THE USE OF THE MANUAL 85

pression or complete disappearance, loses the arbitrary diagnostic character
which furnishes the basis for separating two adjacent groups. In such cases
it has at times seemed more practical to add such a species to the group most
nearly allied to it by general colony aspect, but to cross-reference it to the
related group.

The detection and description of species hitherto unrecognized neces-
sitates extensive review of the literature and restudy of available living
cultures of at least one whole section of the great genus Aspergillus. Unless
the one who encounters a form that he cannot recognize under names
already in the literature is prepared to investigate his form adequately in
relation to the whole genus, he should not describe his organism as a new
species.

Summarized

Comparative study of the taxonomic literature brings out the need for a
standardized series of morphological and descriptive terms into which the
many usages introduced in the two centuries since Micheli can be translated.
Variations in measurement are deemed significant only if they exceed the
common limits between closely related organisms and predominate in the
preparations examined. Ranges in measurements are more significant
than exact dimensions of either selected structures or averaged values based
upon many measurements.

Merely quantitative variations are not recognized as warranting separa-
tion of species. For example, differences in the shade of color, or the in-
tensity of a particular reaction, especially when other strains are found
to fall between the "old" and proposed "new" species, are not regarded as
species characters. Such proposed names either fall to varietal status or to
synonymy.

The names not accepted here fall into several categories. (1) Many are
listed as synonyms because they are believed to have been given to variants
not recognizable by dependable and interpretable differences from other
members of the same series. (2) Fantastic variants, or "monsters",
appearing in culture may, like "Cladosarum", be maintained in the labora-
tory, but unless found perpetuating themselves in nature, clearly fall in the
class of "natures experiments" which do not contribute to the permanent
flora. Such names are not regarded as established. (3) Unidentifiable
species — names appearing in the literature based upon structures or re-
actions regarded by the describer as unique, but whose identity is so com-
pletely lost in large collections among series of closely related strains as to
make them unidentifiable by description — are listed in the check list with-
out characterization.

86 A MANUAL OF THE ASPERGILLI

SPECIES KEYS

No general or comprehensive key to the species of Aspergillus is pre-
sented. Instead, a series of comparatively simple species keys are included
in the discussions of the several groups. The recommended procedure in
identifying an Aspergillus with the aid of this manual is first to determine
its group relationship by means of one or more of the group keys presented
below, then assign the culture more precisely to species by means of the
intra-group species key for the particular group to which the form belongs.

GROUP KEYS

Three keys to the groups of Aspergilli are offered : The first is presented
in the form of a diagram and presents the different groups in what we con-
sider their natural order. Presentation of this key in graphic form, it is
believed, will materially assist the user in grasping the various characters
which ally and interrelate the different groups. Primary separation is
based upon the number of series of sterigmata, whether single or double.
Secondary separation is based upon the character of the conidiophore,
whether rough or smooth. Tertiary separations are based upon the
presence or absence of perithecia, hulle cells, and sclerotia, and upon
the color of the conidiophore wall.

In assigning species to groups by means of this key, the transitional or
intermediate character of certain species becomes strikingly apparent. For
example, Aspergillus caespitosus possesses the brown conidiophore and
conidial coloration of A. nidulans, furthermore, it produces clusters of ir-
regular, thick-walled hulle cells; but the head is radiate, or only loosely
columnar, and no perithecia or ascospores are produced. It is placed in the
A. nidulans group with full recognition that it possesses certain characters
which relate it to A. ustus. Aspergillus alliaceus is another form with
intermediate characters. The conidiophore is uncolored and smooth when
examined in liquid mounts (appearing finely roughened when examined
dry), and the sclerotia are black, but the heads are essentially ochraceous
in color. It is placed in the A. wentii group but shows unmistakable re-
lationship to A. ochraceus. Aspergillus sparsus is a species of uncertain
relationship. It possesses a conspicuously roughened, yellow conidiophore
and globose head, and is placed in the A. ochraceus group; but the conidial
heads show a greenish color which is not found in any other known member
of this group, while the character of the conidiophore will not warrant place-
ment elsewhere. The so-called "bronze series" in the A. tamarii group
is transitional in the direction of A. flavus. Colonies are conspicuously
green when young and retain a greenish tint for a considerable period, in
contrast to A . tamarii which never shows true green and appears greenish
only transiently when young. Such a list of intermediate species and forms

THE USE OF THE MANUAL

87

A.clavotus Group

(Heads clovote)

\

_

.J

A.glaucus Group

(Penthecia yellow)

>

>Ascosponc

^ Single
x Stengmato

A.fumigatus Group

A.nidulans Group

(Ascospores red)

~"\

\

->

V Sclerotia
r Laciung

^Conidiophores
f Smooth

A.ustus Group

IConidiophore

A.flavipes Group

J

Brownish

V huiie
/ Cells

J

A. versicolor Group

A.terreus Group

J

A.candidus Group

>

A. niger Group

(Heads globose, in "block" shades)

V Double

r Slengmata

A. wentii Group

A. tamarii Group

VSclerotia
r Present

VConidiophores
/^ Rough

>

A.flavus-oryzae Group

(Heads yellow— green)

A.ochraceus Group

(Heads ochre to yellow)

>

Comdiophore
> Walls
Yellowish

;

Fig. 20. Graphic representation of natural relationships among groups comprising
the genus Aspergillus. The abundance of species in the different groups is roughly
indicated by the size of the group boxes in the figure.

could be extended, but sufficient have been cited to indicate that the various
groups, like the species which comprise them, often cannot be set apart
by sharp lines of demarcation.

88 A MANUAL OF THE ASPERGILLI

In using this manual, and in studying the Aspergilli generally, it is im-
portant that the worker should realize that these organisms vary within
the species, the species vary within the group, and to a lesser extent, the
groups themselves vary within the genus. In other words, nature did
not realize that we were going to write this manual when the various
species and groups were being developed, hence not all of the forms one
encounters will fit into the various compartments which have been con-
structed, although these are, on the whole, comparatively elastic. This can
be illustrated in another way. If we completely disregard color, the genus
Aspergillus, as depicted in figure 20, can be likened to the spectrum. There
is a green region and a yellow region in the spectrum and the two regions are,
on the whole, distinct. Furthermore, within each of these, certain fixed
and definite lines can be identified. It is, however, extremely difficult, if
not impossible, to say where the green region ceases and the yellow begins.
So it is with the species and groups of the Aspergilli. There is a definite
nidulans group and a definite ustus group, but the line separating the two is
extremely tenuous.

We do not, in any sense, infer that the Aspergilli cannot be classified —
that they cannot be separated into groups, species, and even varieties.
This manual is evidence that they can. But we do wish to emphasize that
we are dealing with living and variable organisms, and that in describing
them, we should be as explicit as possible and still keep our concepts reason-
ably elastic.

Key to Groups — Based Primarily Upon Color

The second key is based primarily upon color and is entirely artificial
in its construction. The various groups are separated by contrasting
coloration, and closely related groups may appear widely separated in the
key. In practice, such a key is very useful since color is the most obvious
character of an Aspergillus, and since the species comprising a particular
group, with but few exceptions, are characterized by variations in shade of
color rather than differences in basic coloration. Presumptive assignment
of the Aspergilli to groups can usually be made from this type of key which
is based primarily on color supplemented by the use of a handlens or dis-
secting microscope.

A. Conidial heads in definitely green, blue-green, or yellow-

green shades in young fruiting colonies B.

AA. Conidial heads lacking green colors (Greenish in excep-
tional cases) K.

B. Conidial heads in green and blue-green shades C.

BB. Conidial heads in yellow-green shades A. flavus group

THE USE OF THE MANUAL 89

C. Conidial stalks and heads coarse— heads clavate A. clavatus group

CC. Heads not clavate D.

D. Colonies mostly showing yellow perithecia and more or

less yellow and red hyphae A. glaucus group

,DD. Colonies lacking yellow perithecia and more or less yel-
low and red hyphae E.

E. Colonies producing columnar spore masses F.

EE. Colonies producing radiate, globose, or hemispherical

heads H.

F. Rapidly growing and spreading colonies G.

FF. Slowly and restrictedly growing colonies A. restrictus series

G. Conidial columns long, narrow A.fumigatus group

GG. Conidial columns short and broad; perithecia usually

present, ascospores red A. nidulans group

H. Heads radiate in blue-green, dull green, to pale tan or

flesh-colored shades A. versicolor group

HH. Heads in some other color L.

K. Heads in long compact columns, avellaneous to cinna-
mon, shading toward colorless through light flesh
colors A. terreus group

KK. Heads in some other color L.

L. Colonies more or less floccose; heads in dull olive-grays

to fuscous A. ustus group

LL. Heads in some other color M.

M. Young heads white or only slightly tinged in age N.

MM. Heads in some other color O.

N. Young heads white, usually in short columns, broad-
ening at apex, often becoming avellaneous in age. . . . A. flavipes group

NN. Heads persistently white, larger heads definitely globose

or radiate A. candidus group

O. Heads in sulphur yellow to ochre shades A. ochraceus group

00. Heads in some other color P.

P. Young colonies showing a greenish color passing into

brown A . tamarii group

PP. Heads not showing greenish Q.

Q. Heads in purple-brown to black shades A. niger group

QQ. Heads in yellowish-brown shades, orange to deep brown

to umber color A. wenlii group

90 A MANUAL OF THE ASPERGILLI

Key to Groups — Based Primarily Upon Morphology

The third key is based primarily upon morphology, with colony color
employed as an accessory differentiating character. This key is also arti-
ficial in construction and is designed to separate the various groups by the
simplest and most direct means possible. Groups naturally related may
or may not appear in their proper sequence. With the data developed
upon the descriptive sheet (p. 82), most of the Aspergilli can be traced
to their proper placement in classification schemes by using the following
key. Natural arrangement is disregarded and the same group is occasion-
ally reached in different places in the key.

A. Species producing perithecia and ascospores B.

AA. Species not producing perithecia and ascospores D.

B. Ascospores colorless C.

BB. Ascospores purple-red A. nidulans group

C. Perithecia white to flesh color, enmeshed in a loose net-

work of colorless hyphae A . fischeri

CC. Perithecia yellow to orange, naked, vegetative hyphae

often showing red to orange granules A. glaucus group

D. Conidial heads cylindrical -clavate; vesicles definitely

clavate A. clavalus group

DD. Conidial heads not cylindrical -clavate E.

E. Colonies showing green or greenish color at some stages

of development F.

EE. Colonies lacking green color P-

F. Conidiophore wall rough or pitted G.

FF. Conidiophore wall smooth H.

G. Colonies green or yellow-green to yellowish A . flavus-oryzae group

GG. Colonies greenish-brown when young, becoming rich

brown or umber in age A. lamarii group

H. Sterigmata in one series I-

HH. Sterigmata in two series K.

I. Conidia elliptical to pyriform J-

II. Conidia spinulose, 2.5 to 4M, globose; chains in com-

pact columns A. fumigatus group

J. Colonies mostly showing yellow perithecia; sterigmata

usually coarse A. glaucus group

J J. Colonies lacking perithecia; conidia in narrow columns. A. reslrictus group

THE USE OF THE MANUAL 91

K. Conidiophores in yellow-brown shades L.

KK. Conidiophores not colored 0.

L. Conidial heads definitely green M.

LL. Conidial heads greenish only when young, then in yel-
low-brown shades A . ustus

M. Ascospores produced— purple-red in color A. nidulans group

MM. Ascospores not produced N.

N. Colonies showing irregularly clustered hiille cells A. caespitosus

NN. Colonies not showing hulle cells: bright green, spreading A. unguis

O. Conidial area in dull green shades (sometimes partially

or completely replaced by tan) A. versicolor

00. Conidial area in blue-greens A. sydowi

P. Conidiophore walls smooth Q.

PP. Conidiophore walls rough V.

Q. Conidiophore walls pale yellow in outer layer; heads

white when young often becoming avellaneous in age. A. flavipes

QQ. Conidiophore walls colorless, or partially yellow-brown

near the head R.

R. Conidial chains in solid columns, compact at base S.

RR. Conidial chains radiate at least in the larger and typical

heads T.

S. Conidial heads in avellaneous shades A. terreus

SS. Conidial heads flesh color (in pinkish shades) A. carneus

T. Heads white or tardily in yellowish shades A. candidus group

TT Heads in darker colors U.

U. Conidial heads globose in purple-brown to black, (rarely

brown or paler) A . niger group

UU. Conidial heads globose in yellowish, yellow-brown, and

dark brown shades A . wentii group

V. Conidiophore walls rough, yellow; heads yellow to

ochre A . ochraceus group

Chapter VIII
THE ASPERGILLUS CLAVATUS GROUP

Outstanding Characters

Conidial heads clavate1, large, pale blue-green.
Conidiophores generally coarse, smooth-walled, uncolored.
Sterigmata in one series.
Conidia elliptical, smooth, comparatively thick-walled.

Group Key

Conidial structures not exceeding 4.0 mm. in length.

Aspergillus clavatus Desm.
Conidial structures often 1 to 5 or more cm. in length.

Aspergillus giganteus Wehmer.

Aspergillus clavatus Desmazieres, in Ann. Sci. Nat. Bot. (2) 2: 71, p. 2,

fig. 4. 1834.

Colonies upon Czapek's solution agar growing rapidly at 20-24° C,
plane or slightly furrowed, in certain strains tending to become floccose
but generally characterized by a surface mycelial mat and abundant erect
conidiophores up to 3.0 mm. in length, bearing large, blue-green, clavate
conidial heads evenly distributed or arranged in more or less well defined
zones (PI. Ill, A and Fig. 21 A); reverse generally uncolored, but becoming
browned in age in some strains; odor strongly foetid in some strains, not
pronounced in others. Conidial heads clavate, large, commonly ranging
from 300 to 400m by 150 to 200m, in age splitting into 2, 3, or more divergent
columns of compacted conidial chains (Fig. 21 B), approximately slate-
olive in color (Ridgway, PI. XLVII). Conidiophores 1.5-3.0 mm. in
length, 20 to 30m in diameter, comparatively thin-walled, smooth, color-
less, gradually enlarging at the apex into a clavate vesicle which is fertile
over an area up to 200 to 250m in length and 40 to 60m or more wide (Fig.
21 C). Sterigmata in a single series, varying in size from 2.5 to 3.5m by
2.0 to 3.0m at the base of the vesicle to 7.0 or 8.0 and occasionally 10m by
2.5 to 3.0m at its apex (Fig. 21 D). Conidia elliptical, comparatively

1 Aspergillus janus Raper and Thorn (see page 187) is characterized by a smaller
clavate vesicle in one of its conidial phases. It is hardly to be confused with the
clavatus group, however, because of the whiteness of its conidial masses, the double
series of sterigmata, and the intermixture of more or less abundant green A. sydowi-
like heads in cultures at room temperature.

92

THE ASPERGILLUS CLAVATUS GROUP

93

heavy walled, smooth, 3.0 to 4.0/x by 2.0 to 3.0/j, ocasionally larger in some
strains and irregular in others.

D

Fig. 21. Aspergillus clavatus Desm. A, Colony growing on Czapek's solution
agar, 10 days, room temperature, X 1.3 (Thorn No. 5169). B, Conidial heads showing
characteristic splitting in age, X 12 (Strain NRRL No. 1823). C, Conidial heads
showing characteristic clavate form of vesicle, X 180 (Strain NRRL No. 6). D,
Portion of a conidial head further enlarged, showing closely packed single series of
sterigmata, X 600 (Strain NRRL No. 6).

Cosmopolitan in distribution and especially common in soil, decaying
vegetation, dung and other materials where active decomposition of nitrog-
enous materials is taking place. Cultures examined include numerous

94

A MANUAL OF THE ASPERGILLI

strains from various parts of the United States, together with isolations
from China, British Guiana, Cuba, Panama and European sources.

The species description as presented is based upon a large number of
closely related strains that have been examined, and is adequately repre-
sented by such specific strains as NRRL Nos. 2, 5, 8, and others.

Upon malt extract agar, details of morphology and colony characteristics
may or may not conform with those listed above. For example, in such
typical strains as Nos. 2, 5, and 8, conidial structures are generally more
abundant upon malt than Czapek's agar and may average as much as 20 to
25 percent larger in size. In other strains such as Nos. 4 and 6, a markedly
different response is noted upon this medium. Conidial heads, although

Q

//

'&

Fig. 22. Aspergillus clavatus Desm. (Strain NRRL No. 1: Thorn No. 107). A,
Colony growing upon Czapek's solution agar, 10 days, room temperature, X 1.3.
B, Conidial heads diminutive and somewhat atypical but showing characteristic
clavate form, X 600.

greatly increased in number, are much reduced in size, with conidiophores
generally 1 mm. or less in length bearing heads only 100 to 125/x long and
proportionately reduced in diameter. In these latter forms, conidia are
somewhat irregular in form and generally larger than in the more typical
strains first considered.

Culture NRRL No. 1 (Thorn No. 107) differs markedly from the species
description in producing deeply floccose colonies (Fig. 22 A) and compara-
tively few spore heads which are extremely variable in size. These range
from very small fruiting structures (Fig. 22 B) borne as branches upon
aerial hyphae to structures arising from the substratum which are charac-
terized by dimensions almost typical of the species. This culture is more
nearly normal upon malt than upon Czapek's solution agar but is unique

THE ASPERGILLUS CLAVATUS GROUP 95

among the strains examined by us and must be considered somewhat
atypical. It deserves particular attention because it was obtained from
the Centraal bureau in Baarn in 1908 as Aspergillus clavatus Desm. and has
undoubtedly been widely distributed by that organization — in fact, it is
quite probable that it has been examined by more investigators than any
other strain belonging to this group. We refrain from using it as a basis
for the species description, however, despite its classic history, since the less
floccose and more heavily sporing strains are so much more commonly
encountered in nature.

Members of this species produce a strong alkaline reaction upon many
culture media and this is usually associated with a strong foetid odor. For
example, when grown upon Czapek's solution agar containing only NaN03
as nitrogen and sucrose as a carbon source, the reaction of typical strains
may reach pH 9.5 or even higher accompanied by a strong odor of tri-
methylamine almost approaching putridity. No other species of Asper-
gillus is known to react in this manner although some strains of A . flavipes
give some suggestion of it. The ability to produce, and more particularly
to withstand strong alkaline conditions undoubtedly accounts for the
common occurrence of this species upon dung and other nitrogen rich
substrata undergoing decomposition.

Aspergillus giganteus Wehmer, in Central, f. Bakt., etc., 2, Abt. 18, No.

13/15: 385. 1907.

Colonies upon Czapek's solution agar growing rapidly at 20° C, charac-
terized by an extensive surface and submerged vegetative mycelium and an
early development of abundant conidiophores 2.0 to 4.0 mm. high, followed
by the subsequent development of less numerous conidiophores ranging up
to several centimeters in length (PI. Ill, B and Fig. 23 A), the latter
strongly phototropic and generally more abundant in marginal areas, com-
monly obscuring the more central mass of short conidiophores ; colonies at
first white, becoming pale blue-green as conidial heads mature ; reverse dull
tan, becoming brown in age; odor none to somewhat foetid in certain
strains. Conidial structures varying greatly in dimensions and falling for
the most part into two general size ranges: (1) conidiophores commonly
2 to 3 mm., rarely exceeding 4 mm. in height, bearing clavate heads 200 to
350/x in length; (2) conidiophores one to several centimeters in length,
bearing heads up to 1 mm. in length. The relative proportions of these
head types is strongly influenced by environmental conditions, and specific
strain characteristics. Conidial heads pale blue-green, in age splitting into
2 or more columns extending the length of the vesicle (Fig. 23 B). Vesicles
consisting of the expanded terminus of the conidiophore, ranging from 100
to 250m by 30 to 50m upon short conidiophores to 400 to 600m by 120 to

96

A MANUAL OF THE ASPERGILLI

180/x upon long conidiophores (Fig. 23 C). Sterigmata in a single series
ranging from 3.0 to 4.0m by 2.5 to 3.0m at the base of the vesicle to 6.0 to

- . •. ■ . «.tJ» *t •* »* 2 . . *• *

- .V »

»■-.*. . i! i •• ' . . "■ ,«» 7 • ;, ,

Fig. 23. Aspergillus giganteus Wehmer (Strain NRRL No. 10). A, Portion of
colony on Czapek's solution agar showing characteristic long conidiophores and
large clavate heads; 10 days, room temperature, X 1.3. B, Portion of colony margin
somewhat enlarged, X 11. C, Single conidial head showing the very elongate vesicle
characteristic of the species, X 130. D, Terminal portion of vesicle showing the
closely crowded single series of sterigmata, X 600.

8.5m by 2.8 to 3.5m at the apex (Fig. 23 D).
walled, smooth, 3.5 to 4.5m by 2.4 to 3.0m-

Conidia elliptical, thick-

Plate III

-v-dd 'u'<l'er lnft !' As?e r(,iUus rIaralus Desm., XRRL No. 8. fi i upper right), Aspergillus giganleus Wehmer,
id x-Diiv inciu?ated at 20° C. in one-sided illumination. C (center left). Aspergillus repens (Cda.)

detfary, NKKL No.20. D (center right ), Aspergillus ruber Bremer, XRRL No. 54. E (lower left), Aspergil-
lus anstelodam, L/Mang.) Thorn and Church, XRRL No. 90. /•' lower right), Aspergillus niveo-glaucus Thorn
and Kaper, M(KL .No. 12, . Figures A and B growing upon standard Czapek's solution agar with 3 percent
sucrose; t to /• , growing upon Czapek's solution agar with 20 percent sucrose. (Color photographs by Haines
-Northern Kegional Research Laboratory. Reproduced through co-operation of Chas. Pfizer & Co, Enc

THE ASPERGILLUS CLAVATUS GROUP 97

The above species description is centered upon strain NRRL No. 10
(Thorn No. 5581. 13A) isolated from Yucatan caves by Prof. F. A. Wolf
(1938). Additional strains examined include isolations from Texas, Illi-
nois, Mexico, Puerto Rico, and strain NRRL No. 1725 (Thorn No. 138)
received from Dr. Westerdijk in 1910 as A. giganteus Wehmer.

In the present treatment we include under the species name A . giganteus
Wehmer all strains which produce conidiophores in excess of 1 cm. in length.
While this may appear somewhat arbitrary, it is done since members of the
A. clavatus group seem to fall into two natural series: (1) those which never
produce conidiophores in excess of 5 to 6 mm. in length irrespective of
environmental conditions or medium composition, and (2) those which
regularly produce few to many very long-stalked fruiting structures under
the usual conditions of laboratory cultivation and examination. To the
first of these series is applied the species designation A. clavatus, to the
second, A. giganteus.

Strains of A. giganteus, like those of A. clavatus differ materially in their
growth and cultural appearances upon different culture media. More
striking, however, is their response to light and temperature. This has
been observed by Wehmer (1907), Wolf (1938) and others, and has been
studied somewhat exhaustively by Webb (1942). Using the Wolf isolate,
Webb found that the production of long conidiophores was favored by
cultivation upon media containing from 1 to 10 percent sucrose and in-
cubation at 20° C. in the presence of light or darkness, whereas heavy
conidial production and the development of short conidiophores were
favored by incubation at 30° C. in darkness. Different strains vary ma-
terially in their cultural appearance upon such standard media as Czapek's
solution agar and can be roughly grouped into three sub-series as follows:
(1) wholly typical strains consistently producing the cultural picture as
defined for the species, (2) strains producing an unusually heavy crop of
short-stalked conidial structures in colony centers, followed by the produc-
tion in marginal areas only of long-stalked fruits typical of A. giganteus,
and (3) rather sparsely growing strains in which there is a general admixture
of short-stalked and scattered long-stalked fruits ranging up to 2 to 3 cm.
in length. The latter group is considered as possibly representing atypical
and somewhat depauperate strains of the first. The second seems to con-
stitute a consistent and fairly well-defined cultural entity, but does not
differ from typical forms sufficiently to warrant separation as a variety.

The validity of the species A. giganteus has been questioned by some
authors. Blochwitz (1929) regarded it as a mutation of A. clavatus and
so designated it in his monograph of the Aspergilli. His view may be
correct. It is our belief, however, that the species should be retained since
forms producing the giant conidiophores noted by Wehmer are repeatedly

98 A MANUAL OF THE ASPERGILLI

isolated from nature, and since no evidence has been presented indicating
that these larger forms arise directly from the smaller and more common
forms. While it is true that conditions can be altered so that A. giganteus
cultures suggest A . clavatus, no one has yet demonstrated that the reverse
can be accomplished.

Group Synonyms

The following names have been proposed for specimens belonging to this
group but without adequate data to warrant recognition as valid species :

A. clavellus Peck, in N. Y. State Mus. Nat. Hist. Rept. 34: 49, PI. 2, figs. 1-5.
1881. Described from cooked squash in New York State. No data is presented which
would warrant separation of this form from A. clavatus Desm.

A. westendorpii Sacc. and March, in Rev. Mycologique 7: 149, 1885, was listed
from cow dung. Correctly assigned to A. clavatus by Lindau, in Deutsch. Krypt.
Fl. Pilze 8: 152. 1907.

A. fusco-cinereus Ellis and Morgan was the name attached to Morgan's jacket
No. 674, showing a very small, clavate aspergillus which has not been collected again,
hence never cultivated. It was probably some member of this group.

A. pseudo-clavatus Purjewitch, in Schrift. Naturforsch. Gesell. Kiev 16 (2): 309,
pi. 12, 1900; see also Sacc. Syll. 16: 1028. The organism in culture was reported as
having both primary and secondary sterigmata in a small-sized "clavatus" type of
head, and perithecia with ascospores which were not adequately described. Until
somebody finds this organism again and reports its cultivation and more complete
description, it will remain doubtful. (See Thorn and Church, The Aspergilli,
p. 100. 1926.)

Occurrence and Economic Importance

Members of the A . clavatus group are quite common in soils and decom-
posing materials characterized by a comparatively high nitrogen content.
They appear to be common upon the dung of various animals and in the
writers' experience have been isolated repeatedly from that of chickens.
While the subject has not been adequately investigated, it is probable that
the ability of members of this group to withstand strongly alkaline condi-
tions enables them to operate successfully as agents of decomposition in
situations where almost all other fungi are eliminated.

Antibiosis

Certain strains of A . clavatus produce substances in the substratum which
are capable of destroying Staphylococcus and other microorganisms. Such
activity was first reported by Weisner in March 1942 (Nature 149: p. 356)
and subsequently by Waksman, Horning, and Spencer in August of the
same year (Science 96: p. 202). To the active substance the latter investi-
gators assigned the name clavacin and noted that it appeared similar to, if
not identical with, that studied by Weisner (1942) to which the designation

THE ASPERGILLUS CLAVATUS GROUP 99

clavatin has since been applied. Additional studies by Waksman and his
co-workers have further defined its action and enlarged the list of bacterial
species inhibited (1942b and 1943). Hooper et al (1944) have demon-
strated the identity of clavacin and patulin, a bactericidal substance ob-
tained from Penicillium palulum by Raistrick and associates (1943) and
reported to be of value in the treatment of the common cold.

Philpot (1943) reported the production of a penicillin -like substance by
a strain of Aspergillus giganteus. In earlier tests by Wilkins and Harris
(1942) this strain had been found to produce a substance active against
Staphylococci.

Chapter IX
THE ASPERGILLUS GLAUCUS GROUP1

Outstanding Characters

Perithecia generally present; yellow, globose to subglobose, thin-walled,
suspended in networks of red or yellow hyphae.
Asci 8-spored, without definite arrangement; usually ripening in 2 to

4 weeks.
Ascospores lenticular, smooth or rough-walled, generally showing an
equatorial line or furrow with or without flanking ridges or crests.
Conidial heads more or less abundant, radiate to somewhat columnar,
typically in some shade of green.
Conidiophores smooth-walled, terminating in dome-like vesicles.
Sterigmata in one series, rather coarse.

Conidia elliptical to subglobose, uniformly and characteristically
roughened.

General Considerations

Aerial hyphae encrusted with yellow, orange, or red granules are abun-
dant in perithecial areas of most of the strains of the group. Both labora-
tory cultures and naturally moldy specimens frequently show this as their
most conspicuous character, one which is readily recognized with the hand
lens. In nature, molds of this group appear as patches of green, yellow,
reddish, or reddish-yellow mold, depending upon the relative abundance of
conidial heads, perithecia, and encrusted aerial hyphae, and especially
influenced by the composition of the substratum.

Representatives of the A. glaucus group are universally distributed in
nature and are significant in the incipient spoilage of many organic ma-
terials useful to man. They occur particularly upon products characterized
by a high osmotic tension such as preserves, jams, cured meats, leather
goods, improperly dried hay, moist grain, and soft woods stored under
humid conditions. The classic habitat is improperly dried herbarium
specimens.

The earliest references to any Aspergilli concern representatives of this
group, for botanists early encountered them upon herbarium material.
Micheli in 1729 used the generic name Aspergillus (rough head) for the
conidial heads, characterized by divergent chains of spores, commonly
present upon such specimens. Later in the century, Wiggers (1780) pro-

1 Abridged from: Charles Thorn and Kenneth B. Raper, The Aspergillus Glaucus
Group, U. S. Dept. of Agr., Misc. Pub. No. 426, Washington, D. C, September 1941.

100

THE ASPERGILLUS GLAUCUS GROUP 101

posed the name Mucor herbariorum for the yellow perithecia found mixed
with the Aspergillus heads, which he regarded as a different mold. In
1809, Link designated the green heads Aspergillus glaucus and the yellow
perithecia Eurotium herbariorum. Half a century later, DeBary (1854)
proved that the Aspergillus heads and Eurotium perithecia were borne
upon the same mycelium, hence were one fungus. Although it could be
maintained that the name Eurotium (designating the perfect stage) should
take precedence over Aspergillus (descriptive of the conidial apparatus),
most recent authors have tended to go back to Micheli and use the name
Aspergillus for the whole group because of the obvious relationship of many
conidial forms for which no perithecia are known.

Laboratory Cultivation

The pattern and size of the ascospore, when present, is especially signifi-
cant in describing species of the Aspergillus glaucus group. Nevertheless,
the conidial apparatus and the vegetative mycelium of particular subgroups
are so important that pure culture under known conditions is always de-
sirable. The character of the colony, as well as the amount of growth, is
strongly influenced by the culture medium, and it is only upon substrata
characterized by a high osmotic tension that typical perithecia and conidial
heads are produced. It should be noted, however, that characteristic
heads and perithecia normally develop, although few in number, in situa-
tions where less concentrated media dry out rapidly, as at the edge of an
agar slant. Colony comparisons for correct identification can best be
made in Petri-dish cultures in which direct observation with the compound
microscope is feasible. Incubation at 22° to 25° C. will permit the develop-
ment of satisfactory colonies for descriptive study, although the optimum
for certain species is above or below this range as will be noted in connection
with these descriptions and in the general discussion on the influence of
temperature on colony growth and development in the genus (p. 45).
Colony descriptions are based upon 3-week old cultures, except as otherwise
stated. For comparative culture the authors have followed Dale (1909)
in using substrata containing high concentrations of sugar. The following
formula is recommended :

Czapek's solution agar with 20 percent of sucrose

Sodium nitrate 3 gm.

Dibasic potassium phosphate 1 gm.

Magnesium sulfate 0.5 gm.

Potassium chloride 0.5 gm.

Ferrous sulfate 0.01 gm.

Sucrose 200 gm.

Agar 15.0 gm.

Dist. water 1,000 cc.

102 A MANUAL OF THE ASPERGILLI

Group Limits and Relationships

Exceptional strains lacking perithecia but presenting conidial morphology
clearly belonging to the A. glaucus group in its strictest sense are occa-
sionally found. In addition, the A. reslrictus series (A. penicilloides series
of George Smith, 1931) shows conidial morphology clearly related to the
group, but differing markedly from the usual types in colony coloration and
in the absence of perithecia. Thorn and Church (1926) considered these
latter types as representing intermediate forms between the Aspergillus
glaucus and A. fumigatus groups. In the present treatment, we include
them as non-ascosporic and for the most part diminutive forms sufficiently
related to the ascosporic species to be included with them in the A . glaucus
group.

Certain other groups of the Aspergilli present characters suggesting those
of the "glaucus" group considered here. The ascospores of A. fischeri
Wehmer (See A. fumigatus group) and of the A. nidulans group (which
see; also Thorn and Raper, 1939) in general resemble those of the A.
glaucus group, but the yellow perithecia suspended by yellow and red
encrusted hyphae do not occur outside of this group.

Group Key (Based Primarily Upon Perithecia and Ascospores)

I. Perithecia present.

A. Ascospores lenticular, 6m or less in long axis.

1. Ascospores with convex faces smooth (or nearly so).

a. Equatorial ridges lacking, furrow absent or showing only as a trace

A. repens series

b. Equatorial ridges low and rounded, furrow broad and shallow

A . ruber series

c. Equatorial ridges thin and flexuous, crestlike (spore resembling

a pulley) A. chevalieri series

2. Ascospores with convex faces rough A. amstelodami series

B. Ascospores lenticular, 6m or more in long axis

Large-spored species or the (E.) herbariorum series

II. Perithecia absent.

A. Colonies predominantly in yellow-orange to brownish shades.

1. Heads approximating those of A. repens A. argillaceus Biourge2

2. Heads proliferating, giving rise to many subheads

A. proliferans G. Smith

B. Colonies in dark green or blue-green shades.

1. Restricted, velvety, with short conidiophores and abundant columnar

heads A . reslrictus series

2. Spreading, floccose, with long conidiophores and globose heads

A. itaconicus Kinoshita

2 Culture distributed by Biourge as a new species. Believed to represent only a
non-ascosporic strain of A. repens, hence not recognized as a valid species by the
writers (see p. 111).

THE ASPERGILLUS GLAUCUS GROUP 103

THE ASPERGILLUS REPENS SERIES

Ascospores lenticular, mostly 4.8 to 5.4/x by 3.8 to 4.4/x, smooth-walled,
with equatorial area rounded or somewhat flattened and occasionally in-
dented showing a trace of furrow, but without crests or ridges.

The Aspergillus repens series as based upon the ascospore described
includes a great number of universally distributed strains which retain
some cultural individuality. Consequently several of them have been
described as species by earlier workers. From comparison of a great series
of these forms, it' seems necessary to bring together under the name A . repens
(Cda.) DeBary, a very considerable number of forms (some of them regarded
as species by others) in which the ascospore is typical for the group and the
colony difference falls within lines of quantitative rather than qualitative
variation.

The following key is offered as a means of separating culturally distinct
strains or groups of strains :

A. Conidial heads large, borne above the surface layer of perithecia and enveloping

hyphae.

1. Heads radiate, long-stalked A. repens (Cda.) DeBary

2. Heads columnar, short-stalked A. dierckxii Biourge3

B. Conidial heads small, enmeshed with the perithecia in a felt of sterile hyphae.

1. Felt orange-yellow, loose-textured, radially wrinkled

A. pseudoglaucus Bloch.

2. Felt yellow-buff, close-textured, plane or nearly so A. profusus Hann3

Aspergillus repens (Cda.) DeBary, in Abhandl I. Senkenberg. Natiirf.

Gesellsch. 7:379. 1870.

Synonyms: A. glaucus var. repens Cda., Icones Fungorum 5: 53, Taf*

II, fig. 27. 1842.
A. scheelei Bain, and Sart., Soc. Mycol. de France, Bui.

Trimest. 28: 257-262, pi. X. 1912.
A. B var. scheelei Bain, and Sart., Soc. Mycol. de France,

Bui. Trimest. 28 : 262-267, pi. XI. 1912.

Colonies upon Czapek's solution agar (3 percent sucrose) restricted,
plane or somewhat wrinkled, forming a rather compact felt (fig. 24 Ai),
with the marginal area near Scheele's green (Ridgway, PI. VI) from de-
veloping heads, older areas yellow-green to greenish-gray and enmeshing
large numbers of aborted perithecia producing few ascospores; normal
perithecia found only when such colonies spread over the bare walls of the
vessel. Reverse in shades of greenish-yellow at colony margin to deep
maroon or almost black in older areas.

3 Species name not recognized as valid by authors of this publication.

104

A MANUAL OF THE ASPERGILLI

Fig. 24. Comparative growth of Aspergillus repens, A. chevalieri, and A. ruber
upon two different culture media; three weeks incubation at room temperature. A,
1 and#,A. repens, NRRL No. 17, upon (1) Czapek's solution agar (3 per cent sucrose)'
and (2) Czapek's solution agar with 20 percent sucrose. B, 1 and 2, A. chevalieri
NRRL No. 78; and C, 1 and 2, A . ruber., NRRL No. 52, upon the same media in similar
arrangement.

THE ASPERGILLUS GLAUCUS GROUP

105

Colonies upon Czapek's solution agar with 20 percent of sucrose spread-
ing broadly and rapidly, plane or slightly wrinkled, orange-yellow, com-
monly characterized by broad zones of dull-green conidial heads (PI. Ill C,
and fig. 25, A and B) ; surface growth consisting of loosely woven hyphae
studded with orange granules enmeshing abundant yellow perithecia above

Fig. 25. Comparative growth of members of the Aspergillus repens series upon
Czapek's solution agar containing 20 percent of sucrose; incubation at room tempera-
ture for three weeks. A and B, Typical cultures of A. repens. C, A. pseudoglaucus,
NRRL No. 40. D, A. pseudoglaucus, NRRL No. 45.

which project abundant conidial heads, the whole colony and especially
the marginal areas and adjacent wall of the culture dish commonly over-
grown by a loose aerial network of hyphae bearing conidial heads and scat-
tered perithecia; reverse varying from yellow-orange to deep maroon.

Perithecia very abundant, borne in loose networks of yellow to orange-
red hyphae (fig. 26 A), yellow, spherical to subspherical, mostly 75 to 100m,

Mi.

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Fig. 26. Marginal areas of colonies of representative species of the Aspergillus
glaucus group showing the relative size and arrangement of perithecia (conidial
heads not in focus): A, Aspergillus repens;B, A. chevalieri; C, A. ruber; D, A. am-
stelodatnt; E, A. echinulatus . Figures X 10; inserts X 30. (Reprinted from Thorn
andRaper, "The Aspergillus glaucus Group," U.S. D. A. Misc. Pub. 426: 1-46. 1941.)

106

THE ASPERGILLUS GLAUCUS GROUP 107

occasionally up to 125/x; asci 10 to 12ju; ascospores lenticular, mostly 4.8
to 5.6/x by 3.8 to 4.4ju, smooth-walled, with equatorial area rounded or
somewhat flattened and occasionally indented showing a trace of furrow
but without crests or ridges (fig. 27 A). Conidial heads abundant, varying
in different strains from 125 to 175m in diameter, consisting of diverging
chains of conidia radiating from a hemispherical vesicular apex of the
conidiophore (fig. 28 A); conidiophores smooth, mostly colorless, 500 to
1,000/x in length, broadening at the apex to a vesicular area, about 25 to
40/i m diameter; sterigmata in one series 7 to 10/z by 3.5 to 4.5/u; conidia
elliptical to subglobose, spinulose, mostly 5 to 6.5/x.

Represented by cultures NRRL No. 12, No. 17, and more than a score
of others included in this study. In this connection it should be noted
that of 37 cultures examined in the present study that produced ascospores
characteristic of the A . repens series, 29 produced colonies and microscopic
details that place them in the species A. repens as described.

This description is manifestly broad enough to include strains approxi-
mating the description given by Bainier and Sartory for Aspergillus scheelei
and Aspergillus B var. scheelei (1912b). Evidently A . scheelei was thought
by the describers to represent a species with somewhat larger ascospores
showing a more definite furrow, whereas Aspergillus B var. scheelei was a
strain with smaller ascospores almost without a trace of furrow. Both
species were described as characterized by the production of a yellow pig-
ment. In the authors' experience, a distinction based upon color is largely
invalidated by variants bridging the whole range from yellow-orange to
deep orange-red and even shades of brown when large numbers of strains
of this series are compared in culture. Strains also vary slightly in the
pattern of their ascospores, some rarely producing spores with a trace of
furrow and others bearing a large proportion with such traces. But among
spores of a single strain limited variation in this character is normally
encountered. Thus the presence or absence of a slight furrow, unless
accompanied by significant differences in morphology or colony character,
would not seem to justify specific descriptions in this series.

A strain designated as Aspergillus dierckxii, presumably by Biourge
but thus far unpublished, was included in Gould and Raistrick's study of
pigment production in the A. glaucus group (1934). As received from
Raistrick's laboratory, this organism (NRRL No. 39) produces colonies
showing no zonate arrangement of conidial heads. Further, heads are
borne on shorter conidiophores than in typical A. repens and consist of
columns of conidia rather than radiating chains. Little or no red color
appears in the colonies or in reverse. Although no other strains showing
exactly these differences have appeared in the authors' collection, separa-
tion as a distinct species is believed unwarranted.

108

A MANUAL OF THE ASPERGILLI

,-. : . SmbE

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I

A

I

^i^^S

w

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ifesi

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1

J

W*Jm

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Fig. 27. Ascospores representative of the four small-spored series of the .Aspe?--
</i7Jus glaucus group. A, A. repens. B, A. ruber. C, A. chevalieri. D, A. amstelodami.
In each species upper left and right and center left spores represent surface, profile
views; center right, surface in face view; lower left, optical section in profile; and
lower right, optical section in face view. (Reprinted from Thom and Raper, "The
Aspergillus glaucus Group," U. S. D. A. Misc. Pub. 426: 1-46. 1941.)

JSSt&-

xttS?" x?' Conidial structures in the Aspergillus glaucus group, X 750: 4. A. revens,
NRRL No. 21; 5, A. echinulatus, NRRL No. 131; C, A. restrictus, NRRL No. 154
ZJ, A. ttacomcus, NRRL No. 161.

109

110 A MANUAL OF THE ASPERGILLI

Aspergillus pseudoglaucus Blochwitz, in Ann. Mycol. 27: 207. 1929;
emend Thorn and Raper, U.S.D.A. Misc. Publ. No. 426, p. 12. 1941.

Colonies upon Czapek's solution agar (3 percent sucrose) restricted in
growth, radiately wrinkled, yellow-green to shades of gray, consisting of a
mixture of small conidial heads, young or aborted perithecia and more or
less colorless hyphae ; reverse orange at center becoming lighter toward the
margin.

Colonies upon Czapek's solution agar with 20 percent of sucrose spread-
ing, strongly wrinkled in a predominantly radiate manner, consisting of a
felt of orange-encrusted hyphae enmeshing abundant perithecia, orange
except at margin where yellow-green predominates from the presence of
small conidial heads admixed with perithecia in the mycelial felt (fig. 25 C) ;
reverse yellow becoming orange-brown or maroon in marginal areas.

Perithecia abundant, spherical to subspherical, mostly 60 to 80m though
occasionally 100m in diameter, yellow, embedded in a felt of orange my-
celium; asci 10 to 12/i in diameter; ascospores lenticular, 4.6 to 5.2/x by
3.6 to 4.0m, occasionally 5.6m m long axis, smooth-walled, with equatorial
region rounded or flattened, without ridges, and with furrow generally
lacking though occasionally showing as a trace. Conidial heads few in
number and generally submerged in the mycelial felt, small, mostly 50 to
75m in diameter but occasionally up to 100m; conidiophores mostly 150 to
300m in length, 5 to 8m at the base, broadening to a terminal vesicle 12 to
20m in diameter; sterigmata in a single series, 6 to 8m by 3 to 4m; conidia
subglobose, delicately spinulose, variable in size ranging from 5.5 to 7.5m
in diameter.

Represented in the NRRL collection by No. 40 received from Baarn
as A. pseudoglaucus Blochwitz and No. 41 received from George Smith as
A . fumigatoides Bain, and Sart.

There is reason to believe that the former culture is directly derived from
Blochwitz 's type. It becomes necessary therefore to emend the description
given by him insofar as the measurements and markings of ascospores and
conidia are concerned. Gould and Raistrick (1934) reported biochemical
data upon a culture, No. A 38, received from Biourge as A. fumigatoides
Bain, and Sart. (NRRL No. 41), which is identical with A. pseudoglaucus
(NRRL No. 40) as sent to the authors by Westerdijk. Obviously the cul-
ture from Biourge is incorrectly named. It does not fit the species descrip-
tion nor the figures of A. fumigatoides (1909) in the size of its conidia, the
character of its perithecial wall, or the pattern of its ascospores. The asco-
spores of A . fumigatoides are shown as roughened over their entire surfaces,
as in A. fischeri, whereas those of No. 41 certainly belong in the A. repens
series. Distinctive colony characters, however, maintained stably through
many transfers, together with the size of the conidial apparatus, warrant
separating A. pseudoglaucus from A. repens and maintaining it as a species.

THE ASPERGILLUS GLAUCUS GROUP 111

A strain, NRRL No. 45, received from Dr. B. O. Dodge and Miss Mar-
jorie E. Swift, of the New York Botanical Garden, in 1931, is characterized
by an intensely wrinkled colony and a further reduction in the size and
number of conidial heads (fig. 25 D). Colonies are dull orange red and bear
abundant perithecia enmeshed in a close felt of sterile encrusted hyphae.
Obviously it should be considered with A. pseudoglaucus.

In cultures received from Baarn (NRRL No. 44) and from George Smith
(NRRL No. 45) as A . profusus Hann (nomen nudum) there is a pronounced
accentuation of the floccose habit already noted in A. pseudoglaucus.
Upon 20 percent sucrose Czapek agar these cultures, which are obviously
duplicates, produce spreading, plane or radiately wrinkled, floccose colonies
consisting of a close felt of light tan to buff -colored hyphae, bearing occa-
sional perithecia and widely scattered conidial heads. The perithecia are
commonly embedded deep within the felt, whereas the conidial heads are
most evident at the colony margin. Although the ascospores of these
strains are definitely of the A. repens type, they are generally flattened
along their equators and commonly show a trace of furrow. An occasional
ascospore shows a minute roughness in the equatorial region. The dif-
ferences observed do not seem to warrant perpetuating the name A. pro-
fusus, and in agreement with Dr. Westerdijk and coworkers (Centraal-
bureau List, 1939), the cultures have been assigned to A. pseudoglaucus.

Culture NRRL No. 46, received from Raistrick in 1923 as Aspergillus
novus Wehmer (nomen nudum) bears ascospores duplicating those of the
cultures just considered. This strain is of particular interest, because in
routine transfers colonies of two distinct types commonly appear. One
of these is predominantly floccose and suggests the colonies of the strains
received as A . profusus. The other consists of a crowded surface layer of
perithecia, which is thinly veiled by a loose felt of orange-red hyphae, and
in its gross appearance, with the exception of its lighter color, is strongly
suggestive of certain cultures of Aspergillus ruber. The authors agree with
Wehmer (1901) and Blochwitz (1929a) that the species designation, Asper-
gillus novus, should be withdrawn.

A nonascosporic culture distributed by Biourge as Aspergillus argillaceus
n. sp., was received upon two occasions from Prof. Raistrick's laboratory.
From its appearance in culture and the morphology of its conidial struc-
tures this fungus would seem to represent a member of the Aspergillus
repens series in which perithecial development has been wholly suppressed.
Although it is questionable whether this fungus represents a true species,
a brief description is given because of its inclusion in biochemical studies
by Raistrick and coworkers (1939, 1934, 1937). Colonies upon Czapek's
solution agar with 20 percent of sucrose spreading irregularly, consisting of
a loose floccose felt of aerial hyphae and abundant conidial heads, pale
yellow-green to clay color; reverse yellow. Upon Czapek's solution agar

112 A MANUAL OF THE ASPERGILLI

(3-percent sucrose) colonies restricted, raised in center, thinning toward
margin, consisting of abundant conidial heads and interlacing hyphae, buff
to clay colored; reverse yellow to tawny. Conidial heads abundant, dull
green, up to 200m in diameter and commonly splitting into fairly well-de-
fined columns, conidiophores up to 1,000m in length. Conidia subglobose
mostly 5.5 to 6.0m but occasionally up to 7.0m in long axis, spinulose.

ASPERGILLUS RUBER SERIES

Ascospores lenticular, 5.0 to 6.0m by 4.0 by 4.8m, colorless, with broad,
shallow furrow generally evident and flanked by low ridges, and with walls
smooth except for minute roughness along the equatorial ridges.

This series includes a great number of strains showing variations in cul-
tural appearance but producing ascospores of a limited size range and
fairly well-defined pattern. For this particular study some 30 strains,
received from various culture collections and contributors and selected
from the isolations made in this laboratory over a period of many years,
have been chosen for repeated culture and examination. Among these,
many strains appear distinct, but their differences are commonly bridged by
intermediate forms. Separation within the series, therefore, must be along
one of the following lines — either (1) strains must be separated upon minor
characters, such as differences in the intensity of pigmentation, slight
variations in ascospore character, etc.; or (2) strains must be set off in
broad and elastic subgroups, in some cases including large numbers which
vary appreciably in detail. The second alternative is desirable, for the
first can lead only to increased hairspliting and end in greater confusion
than that which already exists.

Spieckermann and Bremer's designation Aspergillus ruber (1902) is as-
signed to the series, because its members are predominantly producers of
an intense red pigment and bear ascospores of the general size and pattern
described by these authors. The only other described fungus possessing a
similar ascospore and characterized by its red color is Bainier and Sartory's
Aspergillus sejunctus (1911b).

Although it is now quite impossible to say what particular fungus either
pair of investigators had at hand, both are believed to have worked with
members of the large series now under consideration. Aspergillus ruber
is retained since its description is more adequate for the series in addition
to being the prior species.

To accentuate cultural similarities and differences between the strains
studied, a wide variety of culture metiia has been employed. Based upon
their appearance in culture, the strains fall into a few well-defined sub-
groups (see figs. 29 and 30). The strains belonging to one of these seem

THE ASPERGILLUS GLAUCUS GROUP

113

to represent the fungus described by Spieckermann and Bremer (1902) and
are at the same time most abundant in the entire series, hence are consid-
ered as typical of A. ruber (figs. 29 A and 30 A). The general characters of
the remaining subgroups are listed to show the extreme cultural variation

Fig. 29. Different colony types developed in the Aspergillus ruber series in three
weeks at room temperature upon 20 percent sucrose Czapek agar : A , Typical A . ruber,
NRRL No. 52; B, NRRL No. 70, characterized by thin colonies, with mycelium
largely submerged; C, NRRL No. 65, colony floccose, bearing abundant perithecia
and few conidial heads; andZ), NRRL No. 75, deep floccose colony with very abundant
conidial heads and only scattered perithecia.

that is to be expected within the series, but specific names are withheld,
because to perpetuate or propose such would multiply rather than clarify
the confused nomenclature of this abundant and variable series of organ-
isms.

114 A MANUAL OF THE ASPERGILLI

To illustrate more definitely the variation that occurs, the following
outline of possible lines of separation is inserted:

A. Colonies predominantly perithecial.

1. Colonies red, perithecia abundant in a layer at the agar surface and over-

grown by a felt of red encrusted hyphae

A. ruber (Spieck. and Brem.) Thorn and Church, NRRL Nos. 52, 49

2. Perithecia abundant in a loose, floccose overgrowth of red hyphae as well

as in a layer at the agar surface NRRL No. 65

3. Colonies thin, orange-red, perithecia abundant in old cultures and on very

concentrated media NRRL No. 70

B. Colonies predominantly conidial.

1. Colonies gray, heads long-stalked, perithecia few NRRL No. 75

C. Colonies mixed conidial and perithecial.

1. Colonies orange-red and green, zonate, perithecia borne at the agar surface
and piled in a loose network of superficial hyphae NRRL No. 71

D. Colonies predominantly floccose, colonies red-brown, perithecia and conidial

heads few ^4. lovaincnsis Biourge,4 NRRL No. 76

Aspergillus ruber (Bremer)

Synonyms: A. ruber (Spieckermann and Bremer) Thorn and Church, in

The Aspergilli, 112. 1926.
Eurotium rubrum Bremer, in Zeitschr. f. Untersuch. d. Nah-

rung. und Genussmittel IV. 1901, p. 72, also in Die fett-

verzehr. Organismen in Nahr. u. Futtermitteln, Dissert.

Munster 1902.
E. rubrum Spieckermann and Bremer, in Landw. Jahrb. 31 :

81-128. 1902.
A. sejunctus Bain, and Sart., Soc. Mycol. de France, Bui.

Trimest. 27: 361-367, pi. XL 1911.

Colonies upon Czapek's solution agar (3 percent sucrose) more restricted,
plane (fig. 24 Ci), orange-brown to red-brown in color; perithecia generally
abundant though often abortive; conidial heads pea-green to olive-green,
abundant in some strains, few and largely vestigial in others ; reverse orange-
red to maroon.

Colonies upon Czapek's solution agar with 20 percent of sucrose spread-
ing rapidly and broadly in a regular manner or unevenly, plane, predom-
inantly red, ranging from ferrugineous to morocco red; perithecia very
abundant, borne in a dense layer at the agar surface and largely concealed
within and beneath a close-textured felt of red-encrusted hyphae; conidial
heads projecting above the felt, pale gray-green to deep olive-gray, more or
less abundant, and generally crowded near the center or scattered unevenly

4 Species name not recognized as valid by authors of this publication.

THE ASPERGILLUS GLAUCUS GROUP

115

over the colony (PI. Ill D; figs. 29 A and 30 A); reverse in shades of dark
red-brown.

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Fig. 30. Diagramatic representation of cross sections of different colony types
in the Aspergillus ruber series developed at room temperature upon 20-percent sucrose
Czapek agar, showing relative abundance and disposition of the conidial heads (con)
and perithecia (per), and the amount and character of the mycelium (my) above the
substratum (sub) : A, Typical colony of A. ruber as seen in NRRL No. 52; B-F,atypica
colonies as seen respectively in NRRL No. 71, No. 70, No. 65, No. 75, and No. 76.
Scale approximate. (Reprinted from Thorn and Raper, "The Aspergillus glaucus
Group," U.S.D.A. Misc. Pub. 426: 1-46. 1941.)

Perithecia very abundant, largely enmeshed in a felt at the agar surface
(fig. 30 A), yellow to orange-red, spherical to subspherical, mostly 80 to
120m though occasionally up to 140m in diameter; asci 12 to 15m; asco-
spores lenticular, 5.2 to 6.0m by 4.4 to 4.8m, with furrow generally evident
as a broad and shallow depression around the spore equator, ridges low and

116 A MANUAL OF THE ASPERGILLI

often inconspicuous, walls smooth except for minute roughness along
equatorial ridges (fig. 27 B). Conidial heads generally abundant, numerous
in localized areas or scattered thinly over the colony, pale blue-green,
radiate, 150 to 250m in diameter; conidiophore smooth, colorless to orange-
brown, 500 to 750m in length, broadening to 14 to 16m where it passes into
the subglobose vesicular area of 25 to 35m diameter; sterigmata in a single
series 7 to 9m by 4 to 5m; conidia elliptical to subglobose, closely spinulose,
mostly 5 to 6.5m in long axis.

Aspergillus ruber is represented in this study by NRRL Nos. 52, 53, and
others. Of 31 strains examined belonging to the whole series, 19 showed
colonies and microscopic characters that place them within the species
Aspergillus ruber as described above. Although the majority of strains
belonging to the A . ruber series produce plane colonies as noted in the de-
scription, occasionally strains may produce colonies more or less wrinkled.

Culture NRRL No. 65 (figs. 29 C and 30 D) represents a subseries of
several strains that differ from the above not only in colony character upon
20 percent sucrose Czapek agar, as indicated in the preceding key, but also
in their growth upon media of lower concentration. These grow slowly
and poorly on Czapek (3 percent sucrose), potato-dextrose, and wort
agars, producing small raised colonies of 1 to 2 cm. in diameter bearing
neither normal conidial heads nor perithecia.

Strain NRRL No. 70 (figs. 29 B and 30 C) produces abundant perithecia
only on very dry areas of the substratum in old cultures or on media con-
taining a sucrose concentration of 40 percent or more. In contrast to other-
strains, this fungus grows better upon media containing 4 percent agar than
the usual 1.2 percent agar, further establishing its xerophytic character.

Strains such as NRRL No. 75 (figs. 29 D and 30 E), occasionally en-
countered, are predominantly conidial and characterized by rampant
hyphae bearing abundant conidial heads piled in floccose masses above the
substratum and upon the edges of the culture dish or tube. They thus
produce colonies markedly in contrast with the usual Aspergillus ruber
concept. But the character of their ascospores, together with the occur-
rence of occasional sectors in colonies of these strains showing the usual
mixture of perithecia and conidial heads, relates them definitely with A.
ruber.

Strain NRRL No. 71 (fig. 30 B) represents a subsection of the series
in which conidial heads are abundant and generally arranged in fairly
definite zones and patches with loose clusters of perithecia irregularly and
conspicuously distributed among and above the grouped green heads.
These strains are further characterized by somewhat larger perithecia than
those of NRRL No. 52, being mostly in the range of 125 to 150m in diameter,
and by producing less red color in the colonies and in their reverse.

THE ASPERGILLUS GLAUCUS GROUP 117

Culture NRRL No. 76 (fig. 30 F) is characterized by a close felt of
red-brown hyphae, which completely covers the agar surface and in
which scattered perithecia are borne. Conidial heads are scarce and
largely confined to the colony margin. This strain, which was included in
Gould and Raistrick's study of pigmentation in the Aspergillus glaucus
group (1934), was received from George Smith under the name Aspergillus
lovainensis and attributed to Biourge. Except for its dark color this
fungus in culture bears a striking resemblance to one received from Baarn
as Aspergillus profusus (NRRL No. 44), which showed similar floccose
habits. However, the ascospores of the latter are smaller and less furrowed
and are essentially smooth along the equatorial margin. The degree of
relationship between the two is questionable.

In addition to the ascosporic strains definitely placeable in the series,
George Smith has recently described A. proliferans in which the prolifera-
tion of the sterigmata has become so pronounced as to become the most
conspicuous character, while perithecium formation has been suppressed.
In colony characters, however, it belongs here. This strain diverges
further from the type as described but may be arbitrarily placed here by
the branching of its simplified heads and the size and markings of its
conidia.

Aspergillus proliferans George Smith, in Brit. Mycol. Soc. Trans. 26(1/2):

26, PI. III. 1943.

Colonies on Czapek's solution agar spreading very slowly, with growth
at first largely submerged, then with matted floccose aerial mycelium, white
changing to yellowish shades, sporing tardily, with conidial areas gray-
green, reverse yellowish-brown; on wort agar growing slowly but better
than on Czapek, with mycelium white then yellow and finally orange and
tardy development of gray-green to gray conidial areas, becoming more
deeply floccose in age especially at shallow end of the slope ; reverse yellow ;
normal conidial heads loosely radiate; conidiophore smooth, thin-walled,
usually with one or two septa, 4 to 14m in diameter; vesicles occasionally
almost globose, more frequently obconical or mere broadening of the ends
of the conidiophores, up to about 20m in diameter; sterigmata when normal,
in one series, 8 to 1 lju. by 3.5 to 6m, often elongate, septate and bearing
small secondary heads, frequently resembling heads of monoverticillate
Penicillia, or with upper portion much swollen and appearing almost as
very large, thick-walled conidia with long connectives, up to 20m in di-
ameter, with normal and swollen sterigmata often appearing in the same
head; conidia globose or subglobose, rough, fairly dark-colored, 5 to 9.5m
in diameter; perithecia not found. (Species description after George
Smith.)

118 A MANUAL OF THE ASPERGILLI

Aspergillus halophilus Sartory, Sartory, and Meyer (in Ann. Mycol. 28(3/4): 362-
363, PI. III. 1930) appears from the description based upon colonies grown upon
licorice sticks, to have been some member of this general group. No cultures have
been available for comparison, hence placement near A. proliferous of George Smith
can be only tentative. If placement were to be based upon their figure 12, it might
be a species of Scopulariopsis.

ASPERGILLUS CHEVALIERI SERIES

Ascospores lenticular, mostly 4.6 to 5.0> by 3.4 to 3.8m, occasionally up
to 5.2ju in long axis, with walls smooth or slightly rough, with crests prom-
inent, flexuous, often recurved, and with furrow conspicuous but consisting
more of a trough between extended equatorial crests than a depression in
the spore wall.

Strains belonging to this series show appreciable difference in colony
character and to a limited degree in the surface markings of their asco-
spores. The ascospores of all, however, are characterized by their con-
tinuous, prominent equatorial crests which do not form an integral part
of the spore wall, but extend well beyond the margin of the spore body
proper. To use Mangin's exceedingly descriptive term, they are charac-
teristically "pulley-form."

The following key will serve to differentiate groups of strains within the
series :

A. Ascospore walls smooth.

1. Crests prominent, thin, flexuous, often recurved

A. chevalieri (Mangin) Thom and Church

2. Crests evident, low, usually erect

A. chevalieri var. rnultiascosporus Nakazawa et al5

B. Ascospore walls more or less roughened.

1. Crests thin, flexuous, often recurved; conidia roughened. A. oriolus Biourge5

2. Crests thicker, usually erect: conidia smooth

A. chevalieri var. intermedins, Thom and Raper

Aspergillus chevalieri (Mangin) Thom and Church, The Aspergilli, p. 11.

1926.

Synonym: Eurotium chevalieri Mangin, Ann. des Sci. Nat., Bot. (Ser. 9)
10:361-362, fig. 12. 1909.

Colonies upon Czapek's solution agar (3 percent sucrose) restricted,
plane, closely felted, bluish-gray in center, with typical heads and peri-
thecia largely confined to marginal area (fig. 24 Ci); reverse maroon in
center to orange at margin.

Colonies upon Czapek's solution agar with 20 percent of sucrose growing
best at 30° C. or above, spreading, plane to somewhat wrinkled in central

5 Species name not recognized as valid by authors of this publication.

THE ASPERGILLUS GLAUCUS GROUP

119

area (fig. 31 A) with abundant conidial heads in bli'e-green shades, dis-
tributed evenly over the whole surface or more crowded in localized areas,
projecting above a layer of abundant perithecia enmeshed in orange-red
hyphae at the agar surface; reverse in shades of orange-red to brown, more
intense in center.

Perithecia abundant and closely enmeshed in a felt of orange-red en-
crusted hyphae (fig. 26 B), mostly 100 to 140m, occasionally up to 150^,
globose to subglobose, yellow to orange; asci 9 to 10/z; ascospores lenticular,
4.6 to 5.0/x by 3.4 to 3.8/z, with walls smooth, with equatorial crests prom-
inent, thin and often recurved, and with furrow consisting more of a trough
between parallel crests than an equatorial depression in the spore body

Fig. 31. Comparative growth of members of the Aspergillus chevalieri series in
three weeks at room temperature upon 20 percent sucrose Czapek agar: A, Typical
A. chevalieri, NRRL No. 78; B, A. chevalieri var. intermedins, XRRL No. 82.

(fig. 27 C). Conidial heads abundant, pale blue-green, appearing radiate
from divergent conidial chains, mostly 125 to 175^ in diameter, occasion-
ally larger; conidiophores mostly 700 to 850m in length, enlarging to a
vesicular apex, somewhat globose, 25 to 35/z in diameter; sterigmata in a
single series, closely packed, 5 to 7/x by 3 to 3.5/x; conidia subglobose,
spinulose, mostly 4.5 to 5.5m in diameter.

Aspergillus chevalieri is represented in the present study by cultures
XRRL Xos. 78, 79, and others. The species name is limited to strains
bearing ascospores with smooth walls and prominent, thin equatorial crests
because it is believed that these strains most nearly represent the organism
described by Mangin (1909). When grown upon 20 percent sucrose Czapek
agar, these strains are further characterized by their predominantly orange-

120 A MANUAL OF THE ASPERGILLI

red colonies, pale blue-green conidial heads, and dark-colored reverse.
Within the series, different strains vary in the quantity of conidial heads
produced, e.g., NRRL No. 79 regularly produces an abundance of heads,
NRRL No. 78 relatively few.

Strains of this series are not so commonly encountered as are those of
the A. repens, A. amstelodami, or A. ruber series. And within the series,
strains that conform with the typical species description are relatively less
numerous than in these other series. In the present study 14 strains be-
longing to the A. chevalieri series have been examined and only 6, or less
than half, are wholly representative of the species A. chevalieri.

The series as a whole seems to be relatively unstable, and certain strains
and groups of strains appear transitional between this series and the ^4.
repens series on the one hand and the A. amstelodami series on the other.

Culture NRRL No. 88 received from Baarn as the type of Aspergillus
chevalieri var. multiascosporus Nakazawa, Takeda, Okada, and Simo points
toward the A. repens series. Its ascospores have smooth walls, as in the
typical strains of A. chevalieri, but bear low, erect crests in constrast to
the thin, flexuous crests characteristic of this species. Spores lacking crests
are occasionally seen and these closely resemble A. repens. The colony
upon 20 percent sucrose Czapek agar is definitely of the character of A.
chevalieri. Nakazawa and coworkers (1934) separated it from A. chevalieri
because of its more floccose habit and its more abundant production of
perithecia. The former character is evident in the authors' cultures, but
perithecia are not produced more abundantly than in certain strains en-
tirely typical of A. chevalieri.

Another variation from the typical species is seen in culture, NRRL
No. 87 received from George Smith as Aspergillus oriolus and attributed
to Biourge. The ascospores of this culture (and another that is in the
NRRL collection, No. 81) have crests typical of A. chevalieri, but the spore
walls are finely roughened over their entire surfaces. This character is
suggestive of A. amstelodami, although the roughening of the wall is slight
in comparison with that species. The colony upon 20 percent sucrose
Czapek agar is essentially like that of typical strains of A. chevalieri but is
less red in color and bears fewer conidial heads. Although these cultures
can be distinguished from type they are not recognized as warranting sep-
aration.

The roughening of the ascospore wall is further accentuated in a group
of four apparently similar strains, which it is believed are truly inter-
mediate between A. chevalieri and A. amstelodami. Because their asco-
spores bear crests of the A. chevalieri type and hence appear "pulley-form"
they are retained in the species. However, because they differ from typical
strains in additional particulars, they are considered a new variety, namely
Aspergillus chevalieri var. intermedium.

THE ASPERGILLUS GLAUCUS GROUP 121

Aspergillus chevalieri (Mangin) var. intermedins Thorn and Raper, in U. S.
D. A. Misc. Publ. No. 426, p. 21. 1941.

Colonies upon Czapek's solution agar with 20 percent sucrose differing
from the species in texture and color, and presenting withal a picture
intermediate between A. chevalieri and .4. amstelodami (fig. 31 B). Asco-
spores lenticular, mostly 4.6 to 5.2m by 3.6 to 4.0m, occasionally 5.4m in
long axis, with walls roughened and with prominent equatorial crests.
Conidiai heads dull green, radiate to columnar, mostly 100 to 125m in
diameter, and up to 175m in length ; conidia elliptical to subglobose, smooth-
walled, mostly 3 to 4m in long axis.

Represented in this study by culture NRRL Xo. 82 which was received
from George Smith as No. 107 and bore the following notation: "Isolated
G. S. from cotton yarn, 1927. Close to .4. chevalieri— differs in having
smooth, small conidia and ascospores somewhat larger than type." Dupli-
cated by three additional strains received from European sources.

Aspergillus chevalieri var. intermedins appears to be transitional between
the A. chevalieri and the A. amstelodami series. Such a view is supported
(1) by the pattern of the ascospores, which shows both the extended and
often recurved equatorial crests characteristic of .4. chevalieri and the
rough spore walls of A. amstelodami; and (2) by the coloration of the
colony. Aspergillus chevalieri var. intermedins upon 20 percent sucrose
Czapek agar becomes orange-yellow above and orange to light brown in
reverse, A. amstelodami remains bright yellow with reverse uncolored,
whereas A. chevalieri becomes red in the colony and reverse. The smooth-
ness of conidia in A. chevalieri var. intermedins is a distinctive character and
appears in neither A . chevalieri nor A . amstelodami. Although this variety
from many points of view appears to be a hybrid, proof of such origin is
lacking.

Aspergillus diplocystis (Sartory, Sartory, Hufschmitt and Meyer) Dodge, Med.
Myc. p. 625. 1935. Syn. Eurotium diplocyste Sartory, Sartory, Hufschmitt and
Meyer, in Compt. Rend. Soc. Biol. 104: 881-883. 1930. Not E. diplocystis B. and
Br., Jour. Linn. Soc. 14: 55-56 Tab. 10. 1875.

Characterization: Colonies greenish-yellow, becoming yellow from perithecia.
Conidiophores erect , 50 to 100m high, 3.1 to 3.7m in diameter, membrane thick, hyaline.
Sterigmata confined to a portion of head, 5 to 6.25m by 1.5 to 2.5m- Secondary sterig-
mata small ; conidia spherical, 2.25 to 3.1m in diameter, slightly ellipsoid, green (tendre
to cendre); sterigmata sometimes abortive and proliferous. Perithecia canary
yellow asci 4 to 6m by 5 to 7m, containing 8 ascospores which are ovoid, with a furrow
and two crests, 1.5 to 2.5m by 1.8 to 3.1m-

This description suggests an Aspergillus with the heads approximating .4 . nidulans
and the perithecia of A. chevalieri. Ascospore measurements as reported are appre-
ciably smaller than those of A. chevalieri or any other known species of Aspergillus.
It was described from a case of onychomycosis from the thumb and the great toe.
Tentatively placed in the A. chevalieri series. The name is invalid because of E.
diplocystis B. and Br. 1S75.

122 A MANUAL OF THE ASPERGILLI

ASPERGILLUS AMSTELODAMI SERIES

Ascospores 4.7 to 5m by 3.6 to 3.8^, lenticular, colorless, with equatorial
furrow conspicuous, broadly V-shaped and flanked by broad irregular
ridges, with walls irregularly and unevenly ridged or roughened over the
entire surface.

Included in this series are strains that differ greatly in colony appearance.
However, their close relationship is demonstrated by the similarity in size
and pattern of their ascospores and is further shown by the dark olive-green
color of their conidial heads, the bright yellow color of their perithecia,
and the absence of any red either in the colonies or their reverse.

The following key is designed to show the variation that occurs within
the series and to offer a means of separating strains or groups of strains
that are culturally distinct :

A. Colonies predominantly perithecial.

1. Conidial heads abundant in central area and often in concentric zones

A. amstelodami (Mangin) Thorn and Church

2. Conidial heads widely scattered or lacking NIHIL No. 113

B. Colonies predominantly conidial.

1. Perithecia widely scattered, superficial NRRL No. Ill

2. Perithecia abundant in a felted layer above the conidial heads

A.montevidensis Talice and MacKinnon

C. Colonies very thin, perithecia and conidial heads widely scattered

NRRL No. 110

Aspergillus amstelodami (Mangin) Thom and Church, The Aspergilli, p.

113. 1926.

Synonyms: Eurotium amstelodami Mangin, in Ann. des Sci. Nat., Bot.
(ser. 9) 10: 360-361. 1909.
E. repens var. amstelodami Vuill., Soc. Mycol. de France,
Bui. Trimest 36: 131. 1920.

Colonies upon Czapek's solution agar (3 percent sucrose) restricted, 4
to 6 cm. in diameter, plane or closely wrinkled, yellow to dull yellow-gray
in color from abundant perithecia admixed with sterile hyphae and de-
veloping conidial heads; reverse uncolored, becoming tawny in age.

Colonies upon Czapek's solution agar with 20 percent of sucrose spread-
ing, 8 to 10 cm. in diameter, more or less wrinkled and zonate (PI. Ill E,
and fig. 32 A), perithecia very abundant and clustered in masses forming
a dense layer at the agar surface (fig. 26 D), bright yellow in color, lending
a characteristic appearance to the colony; conidial heads deep olive-green,
abundant in colony center and scattered more or less unevenly over the
whole surface, occasionally obscuring the layer of perithecia beneath.

THE ASPERGILLUS GLAUCUS GROUP

123

Reverse persistent^ yellow under perithecial areas, more or less green
where conidial areas predominate.

Perithecia globose to subglobose, mostly 115 to 140/x in diameter, occa-
sionally up to 160m, not covered by or embedded within a felt of sterile
hyphae (fig. 26 D); asci mostly 10 to 12/z, 8-spored; ascospores lenticular,

Fig. 32. Comparative growth of members of the Aspergillus amstelodami series:
A, Typical A. amstelodami, NRRL No. 90; B, NRRL No. 113, a strain producing
abundant perithecia and few conidial heads; C, NRRL No. Ill, strain producing
abundant conidial heads and very few perithecia; D, A . montevidensis , NRRL No. 108.

4.7 to 5.0m by 3.6 to 3.8/;, with prominent V-shaped equatorial furrow and
broad irregular ridges, and with walls roughened over their entire surfaces
(fig. 27 D). Conidial heads radiate-columnar, mostly 120 to 150m in
diameter, occasionally larger; conidiophores colorless to pale yellow-green,
275 to 350m in length, broadening to 10 to 12m in diameter below the
vesicle; vesicle subglobose, 18 to 25m in diameter; sterigmata about 5 to

124 A MANUAL OF THE ASPERGILLI

6.5/x by 2.5 to 3.5^; conidia finely spinulose, subglobose, variable in size,
ranging from 3.5 to 5.2/u mostly about 4/x in long axis.

Represented in the XRRL collection by cultures Xos. 89, 90, and many
others.

Thirty-two cultures belonging to this series have been examined in the
present study. Included in this number are the authors' own isolates from
a wide variety of sources together with cultures contributed by collabora-
tors in this country and abroad. Of these, more than three-fourths regu-
larly produce colonies conforming with the above description of the species
A. amstclodami. Although wide variation in colony character does occur
within the series, it is obvious that such variations are exceptional rather
than commonplace. Accordingly, it is not believed advisable to assign or
create specific or varietal names for these variations although they differ
markedly from the typical A. amstclodami in gross appearance. An excep-
tion to this policy has been made in the case, of cultures received as A.
montevidcnsis, for reasons that will be considered later.

As indicated in the preceding key to the series, marked variation from
the normal cultural character of .4. amstclodami occurs along certain diver-
gent lines.

Culture XRRL Xo. 113 (fig. 32 B) received from Baarn as Eurotium
repcns (Cda.) DeBary and Wor. var. amstclodami Vuill. (1920) represents
a variation that tends toward an almost complete suppression of the conidial
phase with only an occasional small and atypical head present.

In the opposite direction, culture XRRL Xo. Ill (fig. 32 C) recently
received from Bliss in California (isolated from date fruits) represents a
variation that produces a dense stand of conidial heads and only occasional
perithecia, these being borne above rather than below the layer of crowded
conidial heads.

In contrast to both of the preceding, culture XRRL Xo. 110 isolated
from an old shoe, produces an extremely thin, spreading colony that bears
only widely scattered perithecia or conidial heads.

A fourth distinct variation is represented by culture XRRL Xo. 108
received in 1932 from Talice as A. montevidcnsis Talice and MacKinnon
(1931). This fungus is characterized by an initially strong development of
the conidial phase, and subsequently of perithecia in a felted overgrowth,
which in the colony center more or less obscures the underlying conidial
layer. Perithecia and conidial heads are somewhat smaller than in strains
of A. amstclodami. Although this culture does not differ from ^4. amstelo-
dami more widely than the variations previously noted, since it has an im-
puted pathogenic history and since it has been described and distributed
widely under the name Aspergillus montevidcnsis, it is believed advisable
to retain the name in association with this culture. Accordingly, the
writers include the following emended description.

THE ASPERGILLUS GLAUCUS GROUP 125

Aspergillus montevidensis Talice and MacKinnon, in Soc. de Biol. (Paris)

Compt. Rend. 108: 1007-1009. 1931. emend. Thorn and Raper,

U. S. Dept. of Agr. Misc. Pub. 426, p. 20. 1941 .

Colonies upon Czapek's solution agar (3 percent sucrose) restricted,
radiate sulcate, with zonation evident toward the margin, central area
showing coremia, perithecia few or lacking; reverse and agar very dark,
almost black.

Colonies on Czapek's solution agar with 20 percent of sucrose, spreading,
wrinkled and buckled (fig. 32 D), at first bluish-green from massed conidial
heads, with central area later becoming yellow from developing perithecia
in a more or less tufted overgrowth of somewhat floccose mycelium ; reverse
in yellow-green shades to deep olive in colony center.

Perithecia abundant, of variable size and irregular shape with relatively
few fertile asci and ascospores, late in developing, commonly 75 to 100m
in diameter, occasionally larger; asci 10 to 12/x in diameter; ascospores
lenticular, roughened, with broad and prominent furrow flanked by low
acute and irregular ridges, mostly 4.8 to 5.2m by 3.6 to 4.0m, occasional
spores larger or smaller. Conidial heads very abundant, small, somewhat
columnar, with few conidial chains, mostly 70 to 80m wide, occasionally
up to 100m; conidiophore up to 300 to 350m long, frequently very short
when borne upon the aerial mycelium, broadening to a hemispherical dome-
like vesicular area at the apex; commonly deep green or greenish-brown;
vesicle mostly 15 to 20m in diameter, occasionally larger or smaller; sterig-
mata in one series relatively short and thick, 6 to 7m by 3 to 3.5m; conidia
roughened, subglobose, commonly 4 to 5m by 3 to 4m, occasionally 5.5m
diameter.

Type culture isolated by Talice and MacKinnon from the tympanic
membrane of the human ear (1931). It is carried in the XRRL collection
as No. 108.

LARGE-SPORED SPECIES, OR THE HERBARIORUM SERIES

Under Eurotium herbariorum Lk., Mangin includes all of the members
of the group with ascospores more than 6.6m in long axis (1909). In a
general way this represents a very common usage in older literature begin-
ning as far back as Corda in the 1830's. Because neither measurements
nor markings of the ascospores were given, no one can fix the type of E.
herbariorum. In general, the species in the large-spored group have both
conidia and ascospores definitely larger than those in series already de-
scribed. They become very conspicuous to the collector who finds the
anomalous situation of an overabundance of published names and a dearth
of isolations. Over a period of many years the scarcity of strains isolated
in this laboratory which show ascospores larger than 7.0m leads the authors
to believe that such forms are definitely rare if not abnormal. This obser-

126 A MANUAL OF THE ASPERGILLI

vation is, in effect, confirmed by George Smith (1931). From textiles in
particular he has isolated many small-spored strains but none with large
spores. Possibly the present collection contains as many large-spored
strains as it does because the authors have regarded them as curiosities,
and for that reason retained them, whereas scores of strains of such com-
mon species as A . repens or A . amstelodami have been isolated and forthwith
discarded.

In contrast to the small-spored forms, where complete duplication be-
tween large numbers of isolates is the rule, among the large-spored forms
there is a marked tendency for each strain to present a somewhat different
cultural picture, which is commonly coupled with differences in morphology.
This would suggest that these forms are unstable and variable, but such a
conclusion is refuted by their behavior in culture. To illustrate, culture
NRRL No. 131, a strain of Aspergillus echinulatus, has for 20 years of con-
tinuous culture by the authors retained its distinguishing characters, simi-
larly the single known strain of A. medius has been under observation in
this and European laboratories for more than 40 years without appreciable
change.

Thus the problem of assigning a relatively small number of quite distinct
strains is presented. To describe each of them would merely add to the
confusion already existing, hence they have been grouped somewhat, choos-
ing either historic cultures that have become widely distributed or cultures
of marked individuality as representing specific names. Homogeneity
among the strains brought together is not claimed. The names A. glaucus
and E. herbariorum are not identified with particular organisms in this dis-
cussion.

In setting apart a so-called "large-spored series" the authors do not, in
any sense, wish to imply close relationship or genetic continuity within this
subgroup. Species are grouped together primarily as a matter of con-
venience, and (E.) herbariorum is selected as the series designation primarily
because of Man gin's usage of this species name to cover all of the large-
spored forms.

Key

A. Conidial heads green.

1. Asci ripening within 2 to 4 weeks.

a. Ascospores 6.5 to 7.5ju in long axis A . mangini n. comb.

b. Ascospores 7.5 to 8.5ju in long axis A. umbrosus Bain, and Sart.

c. Ascospores 9.0 to lO.Oju in long axis

A. echinulatus (Delacr.) Thorn and Church

2. Asci ripening slowly, 2 to 3 months, colonies favored by 40 per cent sugar.

a. Ascospores with equatorial ridges and furrow A. medius Meiss.

b. Ascospores usually without equatorial ridges and furrow

A. carnoyi (Biourge) Thorn and Raper

B. Conidial heads white A. niveo-glaucus Thorn and Raper

THE ASPERGILLUS GLAUCUS GROUP 127

Aspergillus mangini (Mangin) n. comb.

Synonyms : Eurotium herbariorum ser. minor Mangin, Ann. des Sci.
Nat., Bot. (ser. 9) 10: 365. 1909.
Aspergillus minor (Mangin) Thorn and Raper, U. S. D. A.
Misc. Publ. No. 426, p. 27. 1941.

Colonies upon Czapek's solution agar (3 percent sucrose) very restricted,
attaining a diameter of only 1 to 2 cm. in 3 weeks, irregular and wrinkled,
cream colored to bluish-brown, conidial heads present or lacking, small
perithecia present or lacking, mostly abortive; reverse uncolored to orange-
maroon.

Colonies upon Czapek's solution agar with 20 percent of sucrose plane
or somewhat wrinkled in the central area, spreading evenly, attaining a
diameter of 8 to 10 cm. in 3 weeks (fig. 33 A), predominantly brick-red in
color becoming maroon in age, perithecia abundant and borne in a close
felt of red-encrusted hyphae at the agar surface, conidial heads few in
number, projecting above the perithecial layer, generally distributed over
the entire colony, but occasionally concentrated in localized areas; reverse
in shades of deep red-brown.

Perithecia abundant, largely embedded in and obscured by a close
mycelial felt at the agar surface, yellow to orange, globose to subglobose,
mostly 100 to 120m in diameter, occasionally up to 150m; asci 14 to 16m;
ascospores lenticular, commonly 6.6 to 7.4m by 5.2 to 5.8m occasionally up
to 7.8m in long axis, finely roughened in the equatorial area, ridges low and
rounded or pyramidal in section, furrow generally definite, shallow but
often steep-sided, V-shaped.

Conidial heads few, generally scattered, projecting above the perithecial
layer, pale blue-green in color, radiate, mostly 150 to 200m in diameter, but
frequently larger; conidiophores smooth, pale to dark brown, mostly 700
to 800m in length, occasionally reaching 1 mm. broadening to 15 to 18m
below the vesicular apex; vesicles subglobose, 30 to 40m; sterigmata in a
single series, 8 to 10m by 4 to 5m; conidia dull green, elliptical to sub-
globose mostly 6.0 to 7.5m and frequently 8.0m in long axis.

Represented by culture NRRL No. 117 isolated from an unpainted
board, as type ; and by several additional cultures isolated in this laboratory.
Culture NRRL No. 115, received in 1937 from Oscar W. Richards, of the
Spencer Lens Company, differs from the type by producing ascospores of
somewhat smaller size and with less evident furrow and ridges. Thus, it
may represent a strain transitional between Aspergillus mangini and A.
ruber. However, it is not sufficiently different from A. mangini, either
culturally or morphologically, to warrant its description as a separate
species or as a distinct variety.

Mangin (1909), in his study of the group, found specimens in his collec-

Fig. 33. Comparative growth of large-spored members of the Aspergillus glaucus
group upon 20 percent sucrose Czapek agar at room temperature : A , A . mangini, NRRL
No. 117, 4 weeks. B, A. umbrosus, NRRL No. 120, 4 weeks. C, A. echinulatus,
NRRL No. 131, 6 weeks. D, A. niveoglaucus, NRRL No. 127, 4 weeks. E, A. medius,
NRRL No. 125, 6 weeks. F, A. carnoyi, NRRL No. 126, 4 months. (Reprinted
from Thorn and Raper, "The Aspergillus glaucus Group," U.S.D.A. Misc. Pub. 426:
1-46. 1941.)

12S

THE ASPERGILLUS GLAUCUS GROUP 129

tion with ascospores less than 7.5/x in long axis, yet larger than those of the
small -spored forms which he described. Having no faith in any of the
descriptions existing at the time, he called the aggregate Eurotium her-
bariorum series minor. The cultures cited above have sufficient common
characters to warrant the belief that they are variants of a common stock
which may be constituted a species aggregate to which we apply the
name Aspergillus mangini.

Aspergillus umbrosus Bain, and Sart., in Soc. Mycol. de France, Bui.

Trimest. 28: 267-269, pi. XII. 1912.

Probable synonyms : A . mutabilis Bain, and Sart., in Soc. Mycol. de

France, Bui. Trimest. 27: 458, pi. XVII. 1911.
A. mollis Bain, and Sart., Soc. Mycol. de France,
Bui. Trimest. 27: 453, pi. XVI. 1911.

Colonies upon Czapek's solution agar (3 percent sucrose) very restricted,
attaining a diameter of 0.5 to 1.0 cm. in 3 weeks, raised, tufted, white to
orange-red, bearing neither perithecia nor conidial heads ; reverse colorless
to orange-brown.

Colonies upon Czapek's solution agar with 20 percent of sucrose, plane
or somewhat wrinkled, spreading evenly or irregularly (fig. 33 B), reaching
a diameter of 8 to 10 cm. in 3 weeks, predominantly vinaceous red to orange-
brown in color, consisting largely of a surface felt of sterile hyphae encrusted
with orange-red granules enmeshing abundant perithecia, occasionally
characterized by a loose floccose overgrowth bearing scattered perithecia,
conidial heads pale blue-green, widely scattered and projecting above the
perithecial layer; reverse in red-brown shades.

Perithecia abundant, yellow to orange, globose to subglobose, largely
embedded in a felt of sterile red-encrusted hyphae at the agar sur-
face, occasionally borne in a loose aerial felt, mostly 120 to 140m in
diameter, rarely up to 175m; asci 14 to 16m; ascospores lenticular, mostly
7.2 to 8.0m by 5.6 to 6.4m, occasional spores up to 8.4m in long axis, finely
roughened to smooth in the equatorial areas, ridges low and generally
rounded, furrow shallow, commonly V-shaped (fig. 34 A); conidial heads
few, scattered, projecting above the perithecial layer, pale bluish-green,
radiate, compact, mostly 175 to 250m in diameter; conidiophores smooth,
colorless to brownish, 700 to 850m in length, broadening to 15 to 20m
below the expanded, domelike vesicular apex; vesicle 25 to 40m in diameter;
sterigmata in a single series, 10 to 12m by 4.5 to 6m; conidia pale green,
elliptical to subglobose, spinulose, mostly 7 to 8m in long axis, frequently
larger.

Represented in the present collection by culture XRRL No. 120 received
in 1925 from Dr. Florence A. McCormick, by European strains, and by
several cultures isolated by the authors.

130

A MANUAL OF THE ASPERGILLI

Fig. 34. Ascospores representative of A, A. umbrosus, and B, A. echinulatus. In
each species upper left, right, and center spores represent surface profile views;
lower left, optical section in profile; lower center, surface in face view; and lower
right, optical section in face view. (Reprinted from Thorn and Raper, "The Asper-
gillus glaucus Group," U. S. D. A. Misc. Pub. 426: 1-46. 1941.)

THE ASPERGILLUS GLAUCUS GROUP 131

Culture XRRL Xo. 123 contributed by Dr. Paul Simonart as an un-
named culture from the Biourge collection differs from the species as above
described by consistently producing ascospores with walls entirely smooth,
whereas in other characters the ascospores duplicate essentially those of
XRRL Xo. 120. Further XRRL Xo. 123 produces colonies of lighter color
than other strains under observation and may, in fact, represent a fungus
comparable to that described as Aspergillus mutabilis by Bainier and
Sartory.

Aspergillus u?nbrosus, A. mutabilis, and A. mollis were described by
Bainier and Sartory (1911c, 1912b) primarily upon the basis of colony color
(pigment production) and conidial apparatus, with the ascopsores of A.
umbrosus recorded as slightly less in long axis (8.0 by 5.6m) than those
of the other species (8.4 by 5.6m)- After careful consideration of the
three descriptions and detailed study of the strains in the authors' posses-
sion showing in general the ascospore described by Bainier and Sartory,
it is believed that they had at hand three cultural variants of the same
species. A . umbrosus is retained as the species designation, as it is believed
that their description of this species more adequately pictures the cultural
and morphological characters of the fungi under consideration than either
of the earlier descriptions, which are left as probable synonyms.

Aspergillus echinulatus (Delacr.) Thorn and Church, The Aspergilli,

p. 107. 1926.

Synonyms : Eurotium echinulatum Delacr., Soc. Mycol. de France, Bui.

Trimest. 9: 266, pi. XIV, fig. III. 1893.
.4. brunneus Delacr., in Bui. Soc. Mycol. France 9: 185, PL

XI, fig. 9, 1893; was described as the conidial stage.
E. verruculosum Vuill., Soc. Mycol. de France, Bui. Trimest.

34:83. 1918.

Colonies upon Czapek's solution agar (3 percent sucrose) very restricted,
1.5 to 3.0 cm. in diameter after 4 weeks, marginal area blue-green from
conidial heads and central portion reddish-brown from an overgrowth of
sterile encrusted hyphae, perithecia lacking; reverse deep orange.

Colonies upon Czapek's solution agar with 20 percent of sucrose slow-
growing, plane or somewhat wrinkled, spreading irregularly, attaining a
diameter of 7 to 8 cm. in 4 weeks (fig. 33 C), commonly mottled in appear-
ance due to the uneven distribution of green conidial heads above the
underlying orange-red perithecial layer; conidial heads bottle-green,
abundant, commonly crowded in localized areas but scattered thinly
throughout the remainder of the colony; perithecia abundant and borne
in a felt of hyphae encrusted with red granules at the agar surface, con-
spicuous where not obscured by massed green heads ; reverse cinnamon to
deep red-brown.

132 A MANUAL OF THE ASPERGILLI

Perithecia abundant, embedded in a looose felt of sterile red hyphae at
the agar surface (fig. 26 E), yellow, globose to subglobose, mostly 125 to
150/x in diameter, and occasionally up to 175m; asci 18 to 22m; ascospores
lenticular, mostly 9 to 10m by 6.5 to 7.5m, occasionally up to 11m in long
axis, conspicuously roughened in the equatorial area, furrow pronounced,
broad, ridges prominent and irregular (fig. 34 B).

Conidial heads densely crowded hi localized areas and scattered through-
out the remainder of the colony, bottle-green in color, radiate, consisting of
relatively few, long, divergent chains of conidia, commonly 250 to 300m
in diameter but often larger or smaller; conidiophores smooth-walled,
colorless to brown shades, commonly 700 to 850m in length, occasionally in
excess of 1 mm., broadening from 5 to 7/z at the base to 15 to 20 /x below the
vesicular apex; vesicle 25 to 35/x in diameter, consisting of a domelike
terminus of the broadening conidiophore ; sterigmata in a single series, not
crowded, bottle-shaped, 12 to 15/x by 5 to 7m; conidia elliptical, pyriform,
or subglobose, echinulate, mostly 8 to 10m hi long axis, commonly larger or
smaller, extremely variable.

Represented by NRRL No. 131 isolated in 1921 from figs received from
California. A subculture of this strain, forwarded by Miss Margaret
Church about 1926, is maintained in the Centraalbureau ; the two lines re-
main identical. No ascosporic stage has in the authors' experience been
found in culture NRRL No. 133 received in 1937 from George Smith as
A. echinulatus Delac, and obtained by him from Biourge, but its conidial
development duplicates NRRL No. 131 and it is apparently correctly as-
signed. Da Fonseca's and the Centraalbureau 's isolations of A. echinu-
latus maintained at Baarn produce somewhat smaller ascospores (8 to 9m
by 6.2 to 7.0m) and conidia than No. 131, but otherwise agree essentially
with the species description as given above. Somewhat further reduction
in ascospore size is seen in cultures NRRL No. 523 isolated from honey,
and NRRL No. 137 received from George Smith and Raistrick as Asper-
gillus mongolicus Biourge (nomen nudum). Although these are less red
in color than No. 131 and appear distinct in culture, the authors do not
feel warranted in separating them as a species or variety, believing that they
represent only variations from the general type designated as A. echinulatus.

Bainier and Sartory described A. disjunclus (1911b) and A. repandus
(1911c) as vigorous species possessing ascospores 11 by 6m and 11.2 by
5 6m, respectively. Authentic cultures of these species are not now avail-
able, but the descriptions as published would seem to place them close to
A. echinulatus.

Probable Synonyms

Several Aspergilli with ascospores ranging near that described for A.
echimdatus appear in the literature. Unfortunately, the details of asco-

THE ASPERGILLUS GLAUCUS GROUP 133

spore markings are not given and there is a dearth of data to identify them.
Some of these names are given :

A. disjunctus Bainier and Sartory, in Soc. Mycol. France Bui. 27: 346-368. PI. X,
XI. 1911. Ascospores described as 11.2 by 5.6m with furrow and crests.

.4. repandus Bainier and Sartory, in Soc. Mycol. France Bui. 27: 463. PI. XVIII.
1911. Ascospores described as 11 by 6m with furrow but no crests.

.4. mencieri Sartory and Flament, in Compt. Rend. Soc. Biol. (Paris) 83: 1114-1115.
1920. Ascospores 10 by 4.7m with furrow and crests.

A. godfrini Sartory and Roederer, in Assn. Frangaise pour l'Avancement des
Sciences, 42nd Session, Tunis 1913. pp. 601-603. 1914. Conidial stage only. This
was growing at blood heat and warmer. Its general description suggests affinity
with the large-spored species of the A. glaucus group. It has not been seen in cul-
ture by us.

Strains with the particular measurements reported for the thiee asco-
sporic species listed above have not come into our collection, yet presum-
ably they and many more with minor variations may be found.

Aspergillus medius Meiss., in Bot., Ztg. 55: (337)-344, (353)-357. 1897.

Colonies upon Czapek's solution agar (3 percent sucrose) very restricted,
0.5 to 1.5 cm. diameter in 6 weeks, tufted, consisting of a dense growth of
yellow-brown hyphae bearing neither conidial heads nor perithecia ; reverse
in shades of yellow-brown.

Colonies upon Czapek's solution agar with 20 percent of sucrose at room
temperature very slow-growing (optimum 20°C. ±), strongly wrinkled,
tufted, irregular in outline, attaining a diameter of 5 to 6 cm. in 6 weeks
(fig. 33 E), colony center deep orange-red becoming yellow to white at the
margin, which is characterized by bundles (becoming branching columns
at lower temperatures) of hyphae bearing dark-green conidial heads in the
manner of loose divergent coremia; perithecia ripening very slowly, ma-
turing ascospores in 2 to 3 months, mostly abortive; conidial heads rela-
tively few (more abundant and larger at lower temperatures) and borne
either in coremiform masses or scattered throughout the colony; reverse in
shades of orange -maroon.

Perithecia scattered, mostly abortive, borne in a dense felt of orange-red
hyphae, very slowly ripening, globose to very irregular in form, extremely
variable in size, rarely attaining a diameter of 125m, containing very few
mature ascospores; asci 18 to 20m; ascopsores sparingly produced, lenticu-
lar, mostly 8.8 to 9.6m by 6.0 to 6.8m, occasionally 10m in long axis, some-
what roughened in the equatorial region, furrow broad and shallow, ridges
prominent, relatively thin and irregular.

Conidial heads deep-green, radiate, compact, and of two types: Small
heads 100 to 150m in diameter borne on loose coremiform columns, and
larger heads 200 to 250m in diameter often scattered throughout the colony,

134 A MANUAL OF THE ASPERGILLI

produced more abundantly at 12° to 15° C. than at room temperature;
conidiophores colorless to brown, mostly 250 to 35(V in length, enlarging
to 15 to 20ju below the vesicle; vesicle subglobose, mostly 30 to 40/i in
diameter; sterigmata in a single series, crowded, short, 7 to 8/t by 4 to 5/x;
conidia green, globose to subglobose, finely echinulate, thick-walled, mostly
8 to 10/x in diameter, but frequently larger or smaller.

Represented in the NRRL collection by culture No. 124 which was re-
ceived in 1924 from Raistrick, who in turn received it from the Centraal-
bureau. It is believed to be Meissner's original strain (1897). Subcultures
of this strain are currently maintained at Baarn and by George Smith in
London. The three lines remain identical as shown by parallel cultures
during recent study.

This fungus is distinguished particularly by (1) its very slow growth
upon 20-percent sucrose Czapek agar at room temperature, (2) its tardiness
in producing perithecia and especially in ripening ascospores, (3) its sparse
production of ascospores, and (4) its formation of aerial hyphal bundles
bearing conidial heads in loose coremiform fashion. Further, it grows
much more rapidly upon Czapek agar containing 40 percent of sucrose than
upon that containing 20 percent, a difference in concentration which does
not materially affect the growth rate of such vigorous species as A. repens
and A. chevalieri. Growth is much more rapid at 20° C. than at 28° to
30° C. (fig. 12). The fungus attains a more favorable form at the lower
temperature, at which there is a heavier growth of mycelium, a more
extensive development of aerial hyphal columns, and a greater production
of conidial heads and perithecia.

Culturally this fungus is easily separated. from all other species of the
group, except possibly A. carnoyi.

Aspergillus carnoyi (Biourge) Thorn and Raper, IT. S. Dept. Agr. Misc.

Pub. 42G, p. 34. 1941.

Colonies upon Czapek's solution agar (3 percent sucrose) very restricted
reaching a diameter of only 3 to 4 mm. in 6 to 8 weeks, thin, white, bearing
neither conidial heads nor perithecia; reverse colorless.

Colonies upon Czapek's solution agar with 20 percent of sucrose at room
temperature extremely slow growing (optimum 18° to 20° C), reaching
a diameter of 7 to 8 cm. in 6 to 8 weeks, irregular in outline, somewhat
floccose, forming a deep felt, bearing abundant perithecia and scattered
conidial heads (fig. 33 F), orange-brown in central area to orange at margin
from abundant perithecia in a loose network of sterile hyphae encrusted
with orange-red granules; reverse in orange-red shades.

Perithecia late in developing, abundant, yellow to orange, globose to
subglobose, mostly 125 to 175/* in diameter but frequently larger or smaller,
borne in a loose floccose felt of sterile brown hyphae; asci 16 to 18ju,

THE ASPERGILLUS GLAUCUS GROUP 135

typically 8-spored, frequently with some or all spores aborted; ascospores
lenticular, variable in size and pattern, mostly 7.2 to 8.4/z by 6 to 6.5^ but
often larger (up to 9.0/x in long axis) or smaller (down to 6.5yu in long axis),
generally smooth-walled but occasionally roughened in equatorial area,
generally rounded but often flattened and occasionally indented, with
ridges wholly absent or indefinite, and with furrow absent or present as a
trace only.

Conidial heads sparsely produced, commonly scattered, dull gray-green,
radiate, compact, mostly 150 to 200/z but often up to 250/x in diameter;
conidiophores smooth-walled, colorless, long, commonly up to 2 mm. in
length, uniform in diameter, 12 to 18m, to just below the vesicle; vesicle
subglobose, 40 to 50^ in diameter and occasionally larger; sterigmata in a
single series, crowded, bottle-shaped, 10 to 12^ by 5 to 6m; conidia globose
to subglobose, echinulate, dull green, mostly 8 to 10^.

Species description based upon culture NRRL No. 126 received in 1937
as Aspergillus carnoyi Biourge from George Smith and by him earlier from
Biourge. Presumably the culture is type, although Biourge 's description
of the species remains unpublished. The culture is distinct, not only differ-
ing in its colony character and in the length of its conidiophores but espe-
cially in the variable character of its ascospores. The majority of spores,
although much larger, resemble those of A. repens, whereas spores with
rough walls are occasionally produced. This culture is, therefore, somewhat
of an exception to the general rule of constancy in ascospore pattern, and
the variability of its spores affords one of the best characters for its identifi-
cation.

Aspergillus niveo-glaucus Thorn and Raper, U. S. Dept. Agr. Misc. Pub.

426, p. 35. 1941.
Synonyms: vl. glaucus mut. alba Bloch., Deut. Bot. Gesell. Ber. 50:
248-256. 1932.
A. glaucus var. albida. Speg., An. del Mus. Nac. de Buenos
Aires 6: 332. 1899.

Colonies upon Czapek's solution agar (3 percent sucrose) very restricted,
1 cm. in diameter after 4 weeks, white to cream, bearing abundant small
conidial heads but no perithecia; reverse colorless to yellow-brown.

Colonies upon Czapek's solution agar with 20 percent of sucrose slow
growing, plane, spreading irregularly, 6 to 8 cm. in diameter after 4 weeks
(fig. 33 D), thinning toward the margin, with mycelium in shades of yellow-
orange becoming cinnamon brown in age, more or less obscured by abun-
dant white heads, and with perithecia abundant, yellow (PI. Ill F), em-
bedded and irregularly clustered in a close felt at the agar surface ; reverse
yellow at margin to deep brown at colony center.

Perithecia abundant, yellow, globose, to subglobose, mostly 100 to 125/x

136

A MANUAL OF THE ASPERGILLI

in diameter, occasionally larger, commonly clustered, borne in an inter-
rupted surface felt of buff to brown hyphae; asci 15 to 17/z in diameter;
ascospores lenticular, mostly 7.2 to 7.8/i by 5.0 to 5.6/i, smooth-walled

«

i

3 it I

•

I

t%> a

\

B

^:D

Fig. 35. Aspergillus glaucus group, conidial heads. A and B, A. niveo-glaucus,
NRRL No. 127: A, surface view showing loose, radiate character of heads, X 35;
B, photomicrograph of the same showing large, globose vesicle fertile over almost
the entire surface, X 600. C and D, Aspergillus restrictus, NRRL No. 154: C, surface
view showing long, columnar heads, X 120; D, photomicrograph of the same showing
characteristic small vesicle fertile on the uppermost surface only, X 600.

except in equatorial area, furrows broad and shallow, ridges prominent,
roughened, rounded or acute and often appearing ragged. Conidial heads
abundant, white (fig. 35 A), often becoming browned in age, radiate, mostly
250 to 300 /x in diameter; conidiophores smooth-walled, colorless to brown,

THE ASPERGILLUS GLAUCUS GROUP 137

mostly 1,000 to 1,500m, rarely longer, broadening to 16 to 20// below the
vesicle; vesicle subglobose, 40 to 50/x in diameter (fig. 35 B); sterigmata in
a single series, crowded, 8 to 10/x by 3 to 4m; conidia elliptical to pyriform,
colorless, spinulose, 6 to 8m in long axis.

The cultural description is based upon NRRL No. 127 of this collection
as type; this is duplicated by XRRL No. 130 received from Baarn in 1938
as Aspergillus glaucus Link mut. alba Bloch. Culture NRRL No. 128,
received in 1935 from George Smith and Raistrick and by them from
Biourge as .4. albidus Speg., differs from type by consistently producing a
greater quantity of conidial heads which are commonly of smaller size;
the ascosporic stage of the two is indistinguishable. Apparently the name
"albidus" is a manuscript use in which Spegazzini's variety (1899) has been
raised to species rank, presumably by Biourge. Blochwitz' mutation albus
assigned to .4. glaucus is untenable, because no definite organism can be
designated as A. glaucus. Furthermore, the specific name albus applied to
an Aspergillus has already been used in another section of the genus.

It is possible that Blochwitz (1932c) is right in regarding this as a muta-
tion, but there is nothing to indicate which particular large-spored form is
the parent species. The strain maintains its identity in culture and hence
must be regarded as a species. Yuill (1939), in contrast, has described
white mutants of A. nidulans and A . fumigatus and has properly designated
them as mutants, for they appeared in cultures under observation and are
known to have been derived from typical chromogenic strains.

THE ASPERGILLUS RESTRICTUS SERIES

Thorn and Church (1926) called this series the "Intermediate Forms"
between the A. glaucus and the A. fumigatus groups. George Smith, wish-
ing to emphasize the resemblance of the conidial apparatus to the monover-
ticillate Penicillia called the lot the A. penicilloides group (1931), thus
suggesting Spegazzini's species as the typical member. However, the
important relationship indicated by the structure of the conidial apparatus
is not with Penicillium but with the A. glaucus group of which these forms
appear to be merely reduced members. All of these forms, like many of
the A. glaucus forms, grow characteristically under conditions of physiolog-
ical drought — represented by their frequency upon mildewed textiles as
studied by Smith (1928) and Galloway (1930) or in concentrated cane
products as reported by Owen (1923). Similarly we have found them in
many situations in which physiological drought is attained by physical
dryness or osmotic concentration attained by the presence of high per-
centage of sugar or sodium chloride.

This natural relationship is on the whole better indicated by accepting
Smith's .4. restrictus as typical, and regarding the other known members

138 A MANUAL OF THE ASPERGILLI

of this series as allied species. To perpetuate an assignment to a subdivi-
sion entitled the Microaspergilli, as suggested by some authors, would com-
plicate nomenclature without compensating values. The series is, there-
fore, keyed as a part of the great A. glaucus group.

Series diagnosis: Perithecia not found. Colonies growing weakly
or restrictedly upon Czapek's solution agar, more freely upon wort agar,
especially well on high concentrations of sugar or salt; green, dark green,
grayish-green, to brownish-green in various strains and under varying
conditions; surface growth consisting of conidiophores only, or of mycelial
felts more or less buckled or heaped; conidiophores smooth, slender, more
or less sinuous, septate; vesicles vary from convex or lens-like areas on the
broadened apices of conidiophores to definitely ovate to globose enlarge-
ments, fertile over all or the upper fraction of such surfaces; heads mostly
definitely columnar, less commonly radiate, hemispherical or almost glo-
bose, especially when young; sterigmata in one series, mostly closely packed
over the fertile area, varying from 2 to 3m by 5 to 6m up to 3 to 4/x by 6 to
10m; conidia barrel-shaped to ovate, mostly in dark greenish shades, smooth
or slowly becoming echinulate or roughened as in the A. glaucus group,
commonly adherent into long chains which are packed into columns.

Series Key

A. Conidiophores broadening upward to produce a convex vesicular apex varying

from 8 to 20m in diameter, producing a long slender column of conidia

A. gracilis Bainier

B. Conidiophores broadening more abruptly, forming a more definite vesicular area.

1. Vesicles more convex, toward hemispherical.

a. Slime development evident A. conicus Bloch.

b. Slime absent or not conspicuous A. restrictus G. Smith

2. Vesicles ovate to hemispherical, columns of conidial chains more or less

conspicuous A . penicilloides Speg.

Differences between the above may appear to be of somewhat minor
character. Nevertheless, each of the sections accounts for sufficient litera-
ture to necessitate separate consideration .

Aspergillus gracilis Bainier, in Bui. Soc. Myc. France, 23: 92, pi. IX, figs.

11-14. 1907.

Colonies on Czapek's solution agar very slow growing, reaching a diam-
eter of only a few millimeters in several weeks (fig. 36A), variously plane
or convoluted or buckled with close textured mycelium at first white, then
slowly green to very dark green, with radiating lines of vegetative mycelium
about the denser area of the colony, growing somewhat better upon wort
agar and upon Czapek's agar containing high concentrations of sugar, re-
verse in yellowish shades, conidial heads in columns up to 200 or 300m long

THE ASPERGILLUS GLAUCUS GROUP

139

by 10 to 20 even 25m in diameter, straight or twisted. Conidiophores
mostly arise as very short branches of aerial hyphae up to 20 to 30/z long or
less commonly up to 100 to 125m even up to 250m and gradually broadening
toward the apex to a vesicular area which is very flat, dome-like, almost the
effect of a truncated cone, 8, 10 to 20m or more in diameter, bearing a single
series of sterigmata, 6, 8, 10 by 2 to 3m, or in particular strains growing out
into little conidiophores producing secondary heads ; conidia at first barrel
form then subglobose about 3m in long axis in Bainier's strain, progressively
larger in related strains. No perithecia found.

The type culture has not been seen. Forms with approximately the
morphology described, however, have appeared in strains NRRL No. 145
(Thorn 4246) from moldy corn; Thorn 4197.3 (culture lost) from Owen in

Fig. 36. A, Aspergillus gracilis, NRRL No. 145, on Czapek's solution agar with
20 percent sucrose, after incubation for 2 weeks at room temperature. B, A.
restrictus, NRRL No. 154, on Czapek's solution agar with 20 percent sucrose after
18 days at room temperature.

the sugar laboratory, New Orleans; and a strain from Thaxter appearing as
a contaminant in a Papulaspora culture. A. gracilis may thus be assumed
to represent a form occasionally found especially in very concentrated
substrata. Biourge later contributed a culture, as type, of A. hypo-
janthinus originally described by him as Penicillium hypojanthinum Biourge
(1923). Three other cultures from Biourge labeled P. (Microaspergillus)
hickeyi, Microaspergillus albo-marginatus, and P. (M.) guegueni (figured in
his monograph, Plate XX, but not described) appear in culture to be only
minor variations of this general form. Later, Biourge sent his undescribed
A . sartoryi. This grew more freely and showed the conidiophore and vesicle
of A. gracilis; conidia were 6 to 7m or greater in long axis, definitely rough
and corresponded almost exactly with a culture received from George Smith

140 A MANUAL OF THE ASPERGILL1

as A. gracilis (XRRL No. 156). Whether further accumulation of strains
will justify giving Biourge's proposed name, A. sartoryi, sectional or varietal
status or merely emphasize the completeness of the series as showing great
variations in structural detail is left uncertain. There does not appear to
be any warrant for preserving A. gracilis var. exiguus Bainier and Sartory
(1912 a). According to the description this variety differs slightly in phy-
siological characters from A . gracilis Bainier.

A. conicus Blochwitz, in Dale, Ann. Mycol. 12: 38. 1914. Previously
described by Dale as a Penicillium in Ann. Mycol. 10: 465. 1912.

See also Thorn and Church "The Aspergilli" p. 125. 1926.

Th? outstanding character of this species is found by microscopic ex-
amination of colonies which become buckled or contorted from a close
felting of mycelia. Relationship back toward the A . glaucus group is found
in the sterigmata, and in the elliptical conidia. Heads differing little in
microscopic structure from A. gracilis are found to be more or less
completely submerged in dark green to almost black slime. Many strains
have been collected from widely separated regions showing all gradations
from a trace of slime only to complete submergence of the heads,
particularly in old cultures.

Dale isolated the strain first described from English soil, and sent dupli-
cate cultures labeled Penicillium sp. to Blochwitz and to Thorn; Thom re-
turned the very brief descriptive note without name as published by Dale
in 1912. Later Blochwitz proposed by letter to her the name only, A.
conicus, which was published by Dale in 1914 without further description.
Later, in his own publication, Die Gattung Aspergillus (1929), Blochwitz
denies the slime development as a character of his organism and redescribes
the species in terms to make A . conicus cover the section of this group re-
presented by Smith's A. restrictus (1931). Since the name was already in
the literature for the slimy series which certainly appears in culture from
widely separated places, it should stand as originally applied. Whether
the slime disappears in some strains long kept on artificial media is not
settled.

In 1928, Biourge contributed under the manuscript name A. cyanogenes
a strain which reproduced the characters given by Thom and Church for A .
conicus. This or a nearly related organism appeared as a contaminant in
several of the cultures received from Biourge. The culture (Thom No.
4733.138) figured in his Monograph (1923) as Xo. 126 in Plate XXI, under
P. glabrum Dale was another. Still another strain from Biourge, labeled
A. viridans differed only in the delayed development of the slimy covering
of the columnar mass of conidia.

Plate IV

A (upper left), Aspergillus restrietus Smith, NRRL No. 154. B (upper right), Aspergillus fumigatus
Presenilis, NRRL No. 163. C (center left), Aspergillus nidulans (Eidami Wint., XRRL No. 192. D (center
right), Aspergillus rariecolor (Berk, and Br.) Thorn and Raper, NRRL No. 195-1. E (lower left), Aspergillus
ustus (Bain.) Thorn and Church, NRRL No. 278. F (lower right), Aspergillus flaripes (.Bain, and Sart.)
Thorn and Church, NRRL No. 295. Figure .4 growing upon Czapek's solution agar containing 20 percent
sucrose and 0.5 percent peptone; all other cultures growing upon standard Czapek's solution agar. (Color
photographs by Haines, Northern Regional Research Laboratory. Reproduced through co-operation of
Chas. Pfizer & Co., Inc.)

THE ASPERGILLUS GLAUCUS GROUP 141

Neill in his study of the Aspergilli of New Zealand (1939) applied the
name A. caesiellus Saito (1904) to what was manifestly some strain near A.
conicus Blochwitz, and then placed the whole group in that species.

A. restrictus G. Smith, in Jour. Text. Inst. 22: IT 15, fig. V. 1931.
Species characterization by George Smith

Colonies growing very poorly on Czapek agar; growing moderately well
on wort agar, dark dull green, gradually turning grey or brownish -grey;
reverse in some cultures uncoloured, in others green to dark green ; surface
velvety at first, becoming wrinkled and often acquiring a warted appearance
(PI. IV A and fig. 36 B); heads forming long, compact, slender columns
(fig. 35 C) up to 350m by 20 to 30m in diameter; conidiophores arising mostly
from substratum but also as branches of aerial hyphae, commonly 50
to 100m, occasionally 150-200^ long by 3 to 3.5m in diameter, often with one
or two septa, smooth, sinuous, uncoloured; vesicles flask-shaped 7.5 to 14/i
in diameter; sterigmata in one series, borne on upper surface of vesicles
only, 6 to 9m by 2.5 to 3m (fig. 28 C); conidia rough, spinulose, elliptical or
somewhat pyriform, often showing a distinct connective, dark greenish-
brown, 4 to 6.5m by 3 to 4m, mostly 4.5 to 6m by 3 to 3.5m; perithecia not
found. The young conidia are hyaline and cylindrical and almost appear
to be segments of enormously elongated septate sterigmata. They
gradually swell without increasing in length, at the same time becoming
pigmented, but even in old heads they adhere strongly together in columns
of parallel chains ; and mounts (fig. 35 D) made in lactophenol usually show
compact, twisted, columnar masses of ripe conidia, both attached to and
separated from the heads.

It is evident from Smith's discussion of a large accumulation of cultures,
especially from the textile industry, that the usual colony-type in his ex-
perience is not the "conicus" type with its slime but the columnar head
described as A. restrictus and accepted here as giving the most appropriate
name to the series.

Among organisms belonging to this series, Smith isolated a single strain
which differed from typical A. restrictus in "showing somewhat larger di-
mensions throughout and, more particularly, in the production of conidia
up to 10 m or more in length," and to which he assigned the designation
Aspergillus restrictus var. B. (G. Smith, Jour. Text. Inst. 22: T115. Figs.
IV, VI, and VIII. 1931). Upon examination by us, this culture (NRRL
No. 148) was found to correspond fairly closely to his published description.
However, we do not believe it is sufficiently distinct to warrant continued
separation as a variety since other strains showing intermediate dimensions
arc encountered.

142 A MANUAL OF THE ASPERGILLI

Aspergillus penicilloides Spegazzini, in Rev. Agrar. Veter.
La Plata, p. 246. 1896.

Spegazzini 's description was emended by Thorn and Church (The As-
pergilli, p. 126, 1926) then broadened by Smith (Jour. Text. Inst. 22, pp.
114-115, 1931) as follows:

"Colonies growing fairly slowly on wort agar, rich dark green with paler
edge, turning darker and duller, and finally becoming dirty greenish-grey
overgrown with sterile hyphae; reverse brown, greenish-brown, and dark
green in patches; surface much wrinkled and folded; heads globose when
young, 40 to 70 /x in diameter, becoming columnar, somewhat ragged, and
up to 200^ long; conidiophores arising either from substratum or from aerial
hyphae, smooth, thin-walled, 75 to 150 m long by 6 to 10 n in diameter;
vesicles rather sharply marked off from conidiophores, pear-shaped to sub-
globose, 15 to 23/xin diameter, fertile over the upper half or two-thirds;
sterigmata in one series, crowded, 8 to 10 n by 2.5 to 3.5 n; conidia ovate,
barrel shaped or nearly spherical, usually showing connective, rough, 3.5
to 5m by 3.2 to 4/x, with very dark colored walls."

Thorn and Church had strain No. 4197.3 isolated from cane products in
Louisiana by Owen and agreed to by Spegazzini; NRRL No. 151 (Thorn
No. 7), also from cane products, fits this description satisfactorily; as did
also Biourge's strain labeled A. pertardus. Smith reports various strains
from mildewed textiles.

It would thus appear that the vesicular area in the series varies from the
curved apex of a clavate conidiophore as in A. gracilis, to a fairly well-
defined hemispherical vesicle as in A. restrictus, and finally to an almost
globose body as in A . penicilloides. All have so much in common with each
other and with the A. glaucus group that their relationship as "degraded"
mutants appears probable.

Aspergillus itaconicus Kinoshita, in Botan. Mag. Tokyo 45: 60-61. 1931.

Probable synonym: A. varians Wehmer, in Bot. Centralb. 80: 460-1.

1899; also in Wehmer Monogr. 77-79, Taf. I, fig.
1. 1899-1901.

Diagnosis from Kinoshita's organism obtained from Dr. Westerdijk.
Colonies on Czapek's solution agar forming dense felts 1 to 2 mm. deep,
white or yellowish, ridged and irregular, with scattered long-stalked green
heads upon dry areas and on the glass ; fruiting more abundantly upon malt
agar, and particularly upon Czapek's solution agar containing 20 percent
sugar; reverse of colony and agar yellow to orange-reddish upon some
media; heads large, light green, globose to radiate, breaking up easily under
the coverglass; conidiophores smooth, colorless, 8 to 16 ju or larger in diam-
eter and up to several millimeters in length under some conditions, with

THE ASPERGILLUS GLAUCUS GROUP 143

walls 0.5 to 1.5m in thickness and splitting lengthwise as A. niger
when broken; vesicles 15 to 4G>, globose or subglobose (fig. 28, D); sterie-
matain 1 series 8 to 9 /x by 1.5 to 2 p\ conidiamore or less pyriform, 4.3 to 5m
by 3.5 to 4/x, finely echinulate, of the A . glaucus type.

Diagnosis is drawn from culture NRRL No. 161 (No. 5344 of Thorn) re-
ceived from Westerdijk as the type of Kinoshita's A. itaconicus and agrees
closely with the description given by Wehmer for A . varians (cf . Thorn and
Church, The Aspergilli, p. 127, 1926), not with Thorn and Church's descrip-
tion of their culture No. 115 which was subsequently lost. A. itaconicus
is so closely related in its physiological responses and in several of its struc-
tural characters to the A. glaucus group that it is placed as an ex-
treme variant at the end of this whole group.

Kinoshita described his organism as a producer of itaconic acid (1931a)
and detailed his experimental work (1931b). The culture as distributed to
laboratories outside of Japan seems to conform to Kinoshita's description
and to produce the acid, but in quantities too small for commercial develop-
ment. More recently, Calam, Oxford, and Raistrick (1939) have recovered
itaconic acid as a metabolic product of A. terreus. From an industrial
point of view, this source appears to be far more promising.

VARIATION IN THE ASPERGILLUS GLAUCUS GROUP

The genetic history of the Aspergilli is an untouched field. Separation
into large groups is easily made definite enough to include all but a few
strains. Within these groups, variation is so great that differentiation of
species requires critical examination and comparison of material, including
extensive culture. Unwilling to undertake this, Neill (1939) disposed of
the whole group with yellow perithecia by calling them all A. glaucus. On
the other hand, Mangin (1909), with the same problem of variability before
him, found the ascospores sufficiently distinctive and dependable to warrant
proposing to separate A . amstelodami and A . chevalieri as separate species
in the A. glaucus group, leaving certain aggregates admittedly inadequately
studied. Bainier and Sartory (1911b, 1911c, 1912), working with members
of these ill-defined species, used color production as the basis for separation.
They cited ascospore measurements as incidental details in description.
Because they appear to have had only a few strains in culture and to have
described them all as new species, the task presented few difficulties to
them; but unfortunately without the original cultures, no one has ever been
able to identify their species with confidence. Raistrick and his colleagues
(1934, 1937, 1938, 1939), using cultures named as received and including
an unpublished series from Biourge, have given quantitiative figures as to
pigment production and pigment mixtures for each culture listed by the
name on the tube, without reporting comparative study of the morphology
found.

144 A MANUAL OF THE ASPERGILLI

It is clear that wide mycelial or colony variations may be found in nature
between strains that retain the ascospore characters of the species. Several
such groups have been held by the authors for 30 years or more and fur-
nish convincing evidence that Mangin was justified in using the ascospore
as the stable and readily determinable integrating character. Some of these
forms retain colony characters in fairly stable form through many transfers
on many laboratory substrata and over a period of years. Others grown in
various substrata in Petri-dish cultures show sectors or other irregularities
in colony habit, which can be picked out and established as strain variants
that maintain their special characteristics in continuous culture.

The possibility that variants of similar nature might be induced by chemi-
cal stimulation led Thorn and Steinberg (1939) to select from the authors'
collection certain strains that had remained fairly constant for many years.
Of this group they subjected strain NRRL No. 90 of A. amstelodami, found
apparently stable for 30 years, to extensive chemical stimulation (1940a).
These experiments yielded two groups of effects: (1) A progressive reduc-
tion in the production of conidial heads and of perithecia, and (2) a great
increase in the mass of vegetative mycelia. In no case was spore produc-
tion completely suppressed although reduced to inconspicuous quantity.
The conidia and ascopsores when examined were found to have retained
the size and markings characteristic for the species, whereas the mass of
vegetative mycelium became excessive and formed a floccose or cottony
mass entirely different in colony appearance from the original.

At this point they reversed the procedure and applied stimulants designed
to reestablish spore production (1940b). As a result, the final cultures
show abundant green heads with normal conidia and numerous perithecia
with ascospores retaining the characters of the species. In routine labora-
tory examination these extreme variants would not suggest the original
strain of A . amstelodami, although both types of variants produce conidia
and ascospores typical of the species.

In 1928, Barnes studied the possibility of heat in inducing variations.
He used a strain reported as "Eurotium herbariorum (Wigg) Link," which
had been isolated and maintained in his laboratory for several years without
apparent changes. Unfortunately, no description of his normal strain was
given, but a strain received from Westerdijk as "Barnes' normal strain"
proves to be identical with A. amstelodami (Baarn strain, NRRL No. 89;
or strain No. 90 as used by Thorn and Steinberg). No reasons were offered
for the original identification.

Barnes described a series of experiments in which the spores of his or-
ganism were subjected to heat under varied conditions, then planted.
From the resulting colonies he described 11 variants, 6 of which are avail-
able in the Centraalbureau. The authors' transfers of these have been

THE ASPERGILLUS GLAUCUS GROUP 145

checked against the descriptions in Barnes' paper and obviously represent
his isolations.

In the following list, his designations appear as quotations, followed by
our identifications based upon careful cultural study:

"Flame" variant is A. ruber.

"Green flame" is A. repens.

"Blue conidial" is .4. chevalieri var. intermedins.

"Creamy" is a pale yellowish strain of Yuills' genus Cladosarum.

"D Brown" is A. uslus.

"C Yellow" is A. amstelodami.
Of these, "C Yellow" (NRRL No. 112) shows the ascosporic pattern and
differs little from Barnes' normal strain or the authors' A. amstelodami.
"Flame" (NRRL No. 59), "Green flame" (culture discarded), and "Blue
conidial" (NRRL No. 85) show ascospores of the glaucus group but differ
markedly in pattern. Among large numbers of induced variations, changes
in the characters of the ascospore have not been found during this study.
"D Brown" (culture discarded) produces no ascospores but develops the
hiille cells and conidial heads characteristic of the Aspergillus ustus group.
These four forms belong to ubiquitous species quite abundant as con-
taminants where plant material is handled. Such contamination is not
satisfactorily excluded by the work reported.

"Creamy" (NRRL No. 143) presents a different problem. The possi-
bility that this " Cladosarum" was actually derived from the "normal" A.
amstelodami is not excluded. Proliferation of the sterigmata in the head of
Aspergilli, especially among the A. glaucus lot, is very common. The
branches produced sometimes are found sterile but usually become diminu-
tive conidiophores with very small vesicles and groups of sterigmata pro-
ducing normal spores. The Yuills' Cladosarum olivaceum (NRRL No. 374)
was found as a conspicuous variant or contaminant in their culture of A.
niger (1938). The colony, conidiophore, vesicle, primary sterigmata, and
initial secondary sterigmata are produced as in A. niger, then instead of
chains of conidia with the newest or youngest conidia at the bases of the
chains and connected directly with the sterigmata, chains of cells are pro-
duced that replicate the sterigmata; occasionally a chain is interrupted by
one cell producing a group of new chains, thus acting as a primary sterigma.
These chains of cells lengthen not at the base as in Aspergillus but at the
distal end. In spite of prolonged search, which shows that the cells toward
the outer ends of such chains lose definiteness as sterigmata, the authors
cannot confirm the finding of a single terminal conidium on each chain as
reported by the Yuills. This morphological picture is repeated by Barnes'
"Creamy" strain, which must therefore be interpreted in terms of Yuills'
genus. Only the two isolations are known thus far. Their failure to pro-

14G A MANUAL OF THE ASPERGILLI

duce true spores and their rapid loss of vitality observed in the present cul-
tures lead to the hypothesis that they are both variants from Aspergillus
and belong to the "monster" type of organisms that fail to survive in com-
petitive environments. Rare occurrences of the "monster" type such as
these can hardly be regarded as permanent members of the fungous flora,
even though the individual can be kept viable by regular vegetative trans-
fer. It is doubted, therefore, whether generic designation is warranted.

Occurrence and Economic Importance

The members of the A . glaucus group are among the most common molds
on earth. Th°y are extremely abundant in nature, and live under all sorts
of conditions, thriving in moist and dry situations. However, they are most
conspicuous upon concentrated substrata, such as drying plant products of
all kinds as they come from the field and reach storage just above the ab-
solute low percentage of water for stability. They are equally com-
monplace in sweetened products, including jams, jellies, soft sugars, honey,
soft candies; in salted products such as meats, pickles, etc.; in dried foods
where preservation is a complex of sugars and acids; upon manufactured
leather goods exposed to humid conditions ; upon clothing and textiles stored
in moist atmospheres; and upon soft wood inadequately cured or subjected
to improper storage. Nevertheless they are frequently overlooked in rou-
tine culture because they grow poorly upon substrata commonly employed
and soon become overgrown by bacteria and more rapidly developing molds.
In following the deterioration of products by cultural procedures, these
forms will appear with amazing frequency if care is taken to select media
approximating the osmotic concentration of the substance examined.

Molds of this group herald incipient spoilage. As excess water increases,
first other molds, then bacteria, appear and complicate the decomposition
process. The presence of any of these organisms is evidence of the earliest
stages of decomposition, and is generally followed by the invasion of other
and more destructive molds and bacteria. Insofar as is known, products
infected by these forms in pure culture are non-poisonous; but naturally
occurring materials in which they appear, or even predominate, should be
considered "suspect" because of the possible presence of toxigenic forms.

Pathogenicity

Members of the A . glaucus group have been reported in connection with
various types of ailments of man and domestic animals. A. hageni of
Hallier (1870) and A. repens have been occasionally reported as fruiting in
the external canal of the human ear; A. montevidensis Talice and MacKin-
non (1931) was isolated from a case of otomycosis ; A . mencieri Sartory and
Flament (1920) was described from sputum of a consumptive; Fonseca

THE ASPERGILLUS GLAUCUS GROUP 147

(1930) isolated, in Brazil, A. amstelodami from a case of mycetoma of the
foot; and .4. keratitis Ball was described as on the cornea (Thorn and
Church, The Aspergilli, p. 86, 1926). There is thus data enough to justify
belief that an occasional strain of the A . glaucus group is found in connec-
tion with lesions in man. We know nothing of the manner of infection and
have no evidence that these organisms appear in any consistent manner as
pathogens. Quevedo's case in which A. maydis (1912) appeared as a cause
of poisoning of horses reads plausibly; this, however, is rendered doubtful
by the observations of other investigators who have reported in widely
separated places such masses of mycelium and spores without encountering
similar injury. A . fontoynonti of Gueguen (1909 and 191 1) isolated from an
abcess was not found pathogenic in subsequent animal experiments.
Species which grow well at blood heat have possibilities of pathogenesis;
fortunately, however, most members of the group do not grow at 37° C.

Chapter X
THE ASPERGILLUS FUMIGATUS GROUP

Outstanding Characters

Conidial heads columnar, in shades of green through dark green to

fuliginous.
Vesicles flask-shaped, typically fertile over the upper half.
Sterigmata in one series, crowded.

Conidiophores smooth-walled, usually colored in shades of green.
Conidia globose, echinulate, green.

Working with lung material from birds dying oi aspergillosis. Presenilis,
about 1850, described and figured the species Aspergillus fumigatus to well
that there has never been any doubt as to the morphology of the mold
present. The investigator of molds in culture, however, quickly finds that
this type of conidiophore and head characterizes not a single pathogenic
strain but a multitude of variant forms that are abundant in soil, upon
decaying vegetation, and. in fact, wherever organic materials are under-
going even the slightest aerobic decomposition. To all oi these forms, col-
lectively, the designation Aspergillus fumigatus group is applied. .

The group falls naturally into two series:

Cultures strictly conidial, varying from velvety to floccose I. fumigatus series

Cultures producing perithecia and ascospores, conidial development generally
limited A. fischeri series

THE ASPERGILLUS FUMIGATUS SERIES

Aspergillus fumigatus Presenilis, in Beitrage sur Mykologie. p. 81, pi. 10,

tigs. 1-11. Frankfurt, 1850 53. Thorn and Church.

The Aspergilli. p. 120. 1926.

Colonies upon Czapek's solution agar spreading broadly over the sub-
stratum, in some strains strictly velvety (PI. IV B and fig. 37 A.), in others
more or less floccose with varying amounts of tufted-aerial mycelium to deep
felted or extremely floccose forms (fig. 37 B), white at first, becoming green
with the development of heads but varying considerably in the final shade of
green, often becoming dark green to almost black in age. Reverse and
substratum, in some strains uneolored. in others showing varying amounts
of yellow, or again passing over in dark red shades in age. Conidial heads
columnar, compact, varying in measurement from strain to strain up to 400
by 50m, hut usually much shorter, occasionally very small. Conidiophores

14>

■f '

Fig. 37. Aspergillus fumigatus group. A-C, A. fumigatus: A, Typical, heavy
sponng strain, XRRL No. 178, on Czapek's solution agar, 10 days, room temperature;
B, Floccose, light sporing strain of same species, XRRL Xo. 171 ; and C, Photomicro-
graph of typical conidial heads showing characteristic form of vesicles and crowded
stengmata in a single series, X 500. D-F, Aspergillus fischeri: D, Strain XRRL
Xo. 186, growing upon Czapek's solution agar, characterized by very abundant
perithecia and few conidial heads; E, Portion of a colony enlarged showing crowded
penthecia more or less obscured by loose enveloping hyphae, X 6.0; and F, Asci con-
taining ascospores at various stages of maturity, X 500.

149

150 A MANUAL OF THE ASPERGILLI

short, smooth, usually densely crowded, up to 300m (in occasional strains
up to 500^) m length by 2 to 8m in diameter, frequently more or less green
colored, especially in the upper part, arising directly from submerged
hyphae or as very short branches from aerial hyphae, septate or unseptate,
gradually enlarging upward and passing almost imperceptibly into the
apical flask-shaped vesicles. Vesicles up to 20 to 30m in diameter, usually
fertile on the upper half only (fig. 37 C). Sterigmata in one series, usually
about 6 to 8m (varying from 5 to 10m) by 2 to 3m, crowded, closely packed
with axes roughly parallel to the axis of the conidiophore. Conidia dark
green in mass, echinulate, globose, mostly 2.5 to 3m in diameter with ex-
tremes ranging from 2 to 3.5m. Sclerotia or perithecia are not found. The
species grows well at temperatures up to 45° C. or even higher, and is com-
monly present in compost and other material undergoing decomposition at
high temperatures.

The species description presented is a composite rather than an exact
citation of detailed data about one strain, but NRRL No. 163 (Thom No.
118) may be considered typical. Organisms coming within this series as
described are world-wide in distribution and omnivorous in habit. They
are regularly abundant in soil and in decomposing organic masses ; they are
recoverable from apparently sound cereals, corn, oats, wheat, etc. ; they are
encountered as pathogenes in the air passages of birds and occasionally as
lung parasites of mammals, including man. One strain (NRRL No. 164)
was named A. cellulosae by Hopffe (1919) because of its ability to break
down cellulose, but when examined, it presented no morphologic differences
from hundreds of common isolates from soil.

Efforts to induce perithecium formation by the conidial strains of A.fumi-
gatus have been disappointing. Organisms maintained in culture over long
periods and subjected to multitudes of transfers upon all sorts of substrata
have failed to give any response suggesting perithecium formation.

Extremes of variation in different strains range from colonies character-
ized by crowded conidiophores rising vertically from submerged hyphae to
a height of 300m, or perhaps at times 500m, then producing columns of co-
nidia sometimes up to 400 by 50m, to very floccose forms in which spore for-
mation is generally retarded and in which conidiophores develop as very
short branches of aerial hyphae and produce short columnar heads. Many
of these variants can be isolated and maintained in culture, thus they have
been made the types of species by earlier workers. Others encountered
upon unique substrata have been given specific names in the belief that the
substratum relation was obligate. Yuill (1939) isolated a buff-colored
mutant from a typical green strain and applied to it the designation A.
fumigatus var. helvola (fig. 17 C). Shortly thereafter Steinberg and Thom
(1940) likewise recovered an essentially uncolored mutant from a typical
green strain. Both forms still remain unchanged in culture.

THE ASPERGILLUS FUMIGATUS GROUP 151

If the taxonomist could seize a half dozen widely spaced variants and
destroy those which bridge the gaps between, identification as separate
species would be easy. This, however, becomes impossible when large
numbers of isolates are cultivated and studied in comparative culture. In
attempting to divide this great series into tangible entities, earlier workers
have created a long list of species and, while we do not consider these to be
valid, they are presented in alphabetical order with the places of description.
It is believed that all of these should be regarded as synonyms of A . fumi-
gatus Fres.

A. aviarius Peck, in N. Y. State Museum Rept. 44, p. 25, pi. 4, figs. 9-12. 1891-

A. bronchialis Blumentritt, in Ber. Deutsch. Bot. Ges. 19: 442-446, PI 22; figs 1-A-
1901; also ibid. 23: 419-427, PI. 19, figs. 1, 3, 6, 7, 8, 19, 23. 1909.

A. calyptratus Oudemans, in Arch. Neerl. Ser. II. 7: 283. Tab. XIII. 1902.

A. cellulosae Hopffe, in Centralb. f. Bakt. etc., Abt. 83: 531-537. 1919.

A. desseyi Spegazzini, in Physis (Rev. Soc. Argentina Cien. Nat.) VIII: 115-117,
1 figure. 1925; review only seen, Rev. Appd. Mycol. 4: 542. 1925.

A.fumigatus var. alpha Sion and Alexandrescu, in Compt. Rend. Soc. Biol. (Paris)
64: 288-289. 1908.

A.fumigatus var. minimus Sartory, in Bui. Acad. Med. Paris 3 Ser. 82: 304-305.
1919.

A. fumigatus var. lumescens Blumentritt, in Ber. Deut. Bot. Ges. 23: 419-427,
PL 19, figs. 5, 6, 18-21. 1905.

A. glaucoides Spring, in Bull. Acad. Sci. Belg. 19: 560-572. 1852.

A. lignieresi Cost, et Lucet, in Ann. Sci. Nat. Ser. 9, II: 119, tab. 5, fig. 18-23.
1905.

A. nigrescens Robin, in Histoire Naturelle des Vegetaux Parasites, p. 518, Paris.
1853.

A. pulmonum hominis Welcker, in Kuchenmeisters Parasiten II, p. 144. This is
discussed and figured by Theodor von Dusch, in Virchow's Archiv. (n. f. 1) 11: 561-
566. 1857, but no ground is given for separating it from A.fumigatus.

A. ramosus Hallier, in Zeitschr. Parsit. 2: 266-269, pi. 6, figs. 1-6. 1870.

A. syncephalis Gueguen, in Les Champignons parasites de l'homme et des animaux
299 pp., 12 pi. Paris, 1904.

A. virido-griseus Cost, and Lucet, in Ann. Sci. Nat. Bot. IX. 2: 140. 1905.

THE ASPERGILLUS FISCHERI SERIES

Aspergillus fischeri Wehmer, in Centralb. f. Bakt., etc., 2 abt., 18:

390-2, figs. 1-5. 1907.

Synonyms:^!, fumigatus — ascosporic, see Thorn and Church, in Am.
Jour. Bot. 5: 91-92. 1918. See also Thorn and Church,
The Aspergilli, p. 132. 1926.
Sartorya fumigata Vuillemin, in Compt. Rend. Acad. Sci.
(Paris) 184, No. 3: 136-137. 1927.

The conidial form is strikingly similar to Aspergillus fumigatus. Colonies
grow well upon Czapek's solution agar with conidial heads sparingly pro-

152 A MANUAL OF THE ASPERGILLI

duced at room temperature (fig. 37 D), but more abundantly at 37° C.
Conidial heads frequently small and generally of a lighter green color than
those of typical A. fumigatus. Perithecia quickly and abundantly pro-
duced in 'most strains dominating the colony appearance (fig. 37 E),
commonly up to 300M in diameter, not colored, or very pale salmon, with
walls scarcely colored, consisting of a single layer of cells, crushing easily,
covered by a loose network of uncolored sterile hyphae. Asci abundant,

Fig. 38. Ascospores of Aspergillus fischeri NRRL No 181 (- Thorn No 4651 2)
Upper left, center, and right represent surface, P^.^f^je^fiSKon
section in profile; lower center, surface in face view; and louer right, optical section

in face view.

8-spored, filling the perithecium within a few days, 8 to 10/x by 10 to 12/z
subglobose (fig. 37 F), breaking down quickly to leave the perithecium full
of ripe ascospores. Ascospores biconvex, uncolored, usually about 7 by 4/x,
consisting of a central body 5 by 4M, with two frilled equatorial banks about
1M in width, roughened with echinulations or anastomosing bands on each
convex surface (fig. 38), separating into two valves in germination.

Culture NRRL No. 181 (Thorn No. 4651.2), received as type from
Wehmer in 1923, is apparently identical with numerous isolations rom
American sources. Many strains have been seen, including a series from
sputum of human cases showing lung involvement by X-ray examination.

THE ASPERGILLUS FUMIGATUS GROUP 153

It is regarded as world-wide in distribution but seemingly not abundant
anywhere. The species grows well at temperatures of 37° C. and higher.
Some additional perithecial forms have been described with characters
suggesting relationship with A. fischeri. However, these are inadequately
known in culture and assignment here must remain somewhat provisional.

A. rnalignus Lindt, in Arch. Exp. Path. Pharmakol. 25: 257-271, fig. 1-11. 1889.

Lindt probably had before him some strain which corresponded closely with
Wehmer's Aspergillus fischeri despite the fact that his description of the conidial
apparatus offers sufficient contrast as to lead to question.

A.fumigatoides Bainier and Sartory, in Bull. Soc. Myc. France 25: 112, pi. 5. 1909.

While the describers believed this strain close to A. fumigatus, Thorn and Raper
(1941) found it necessary to place the organism studied under this name by Gould
and Raistrick (1934) in A. Pseudoglaucus, although the original description and
figures of Bainier and Sartory probably represented material close to, or identical
with, A. fischeri.

Sartorya fumigaia (Fres.) Vuill., in Compt. Rend. Acad. Sci. (Paris), 184(3): 136-
137. 1927.

Sartory, Sartory, and Meyer (1926) reported that a culture of A. fumigatus sub-
jected to radiation produced an ascosporic form. Later this was designated by
Vuillemin (1927) as a new genus Sartorya. Neither Vuillemin, nor Sartory, Sartory,
and Meyer appear to have known the relation between A. fumigatus and A. fischeri
although it was pointed out by Thorn and Church in 1918. Since the material named
Sartorya does not appear to have been distributed or fully described, Sartorya fumi-
gate (Fres.) Vuillemin may be regarded as a synonym for A. fischeri Wehmer and the
generic name dropped.

It is believed probable that Aspergillus fischeri Wehmer represents the
primary organism of this group and that from it the conidial form A. fumi-
gatus developed as a species lacking entirely the ascosporic phase. The
fact that A . fumigatus was described first merely reflects the much greater
abundance of this species.

Occurrence and Economic Importance

Aspergillus fumigatus is an extremely cosmopolitan mold and occurs with
particular frequency in soil containing appreciable organic materials, upon
vegetable matter undergoing slow decomposition, and upon imperfectly
dried, stored grains. The mold is an important agent in many decomposi-
tion processes, particularly at temperatures above 37° C. Growing success-
fully at 45° to 50° C, within the lower reaches of thermophilic decomposi-
tion, it is able to operate within a range where most fungi are excluded.
Whereas some forms have been described as very active agents of decom-
position (e.g. A. cellulosae of Hopffe, 1919), their more significant role is
believed that of forerunners of active bacterial decomposition on the one
hand, and as slow destroyers of more resistant tissues on the other. Asper-
gillus fischeri, though much less abundant, may be found in situations
generally similar to those yielding A. fumigatus.

154 A MANUAL OF THE ASPERGILLI

Pathogenesis

There is an extensive pathological literature which covers the occurrence
of A. fumigatus in lesions of birds and mammals, including man, together
with biochemical and animal experimentation. Such experiments have
repeatedly proved that the organism is pathogenic to fowls confined in con-
gested quarters in which moldy grain, straw, and other plant remains are
abundant. Direct inoculation to the cornea in laboratory animals causes
lesions characteristic for the species.

Infection of human beings occasionally appears, and observations seem
to indicate that the patients generally have been exposed to air carrying
large numbers of spores. Allergists have reported asthmatic conditions
arising from sensitization to this species, and Bern ton (1930) reports having
successfully treated a patient by means of an extract prepared from the
spores and mycelium of A. fumigatus. The occurrence of A. fischeri in
cases grouped with A. fumigatus is clear indication that the pathogenic
principle, whatever it is, is generally present in the group although it may
vary in its intensity among different strains as indicated by workers such
as Costantin and Lucet (1905).

Among organisms known to be, or believed to have been, pathogenic
strains of A. fumigatus, the following named forms may be cited: A.
gratioti, A. malignus, A. fumigatoides, A. virido-griseus, A. bronchialis, A.
glaucoides, A. nigrescens, A. pulmonum hominis, A. ramosus, and A. avia-
rius (see p. 151). For a more complete discussion of this group in relation
to disease in birds and mammals, the reader is referred to Thorn and
Church's The Aspergilli (1926) and Dodge's Medical Mycology (1935).

Antibiosis

Anslow and Raistrick (1938a) reported the production by Aspergillus
fumigatus of a substance to which they applied the name, fumigatin, and in
the same year (1938b) reported the species to produce a second metabolic
product termed spinulosin, which they had previously isolated from Peni-
cillium spinulosum. In 1942 fumigatin was further discussed by Oxford
and Raistrick as a powerful agent against such bacteria as Bacillus anthra-
cis, Escherichia coli, Salmonella typhi-murinum, Staphylococcus albus, S.
aureus, Streptococcus viridans, and Vibrio cholorae. Also in 1942 Waksman,
Horning, and Spencer reported an antibiotic substance, termed fumigacin,
to be produced by A. fumigatus, and presented methods of differentiating
this from fumigatin as studied by Raistrick and associates. Fumigacin was
found to be both bactericidal and bacteriostatic in its action and to be
effective in fairly high dilutions in inhibiting the growth of gram -positive
cocci and bacilli ; it was much less effective against the gram-negative mem-
bers of the coli-aerogenes group. Both fumigatin and fumigacin have been
found toxic to experimental animals.

Chapter XI
THE ASPERGILLUS NIDULANS GROUP*

Outstanding Characters

Conidial heads short columnar, usually dark green with primary and
secondary sterigmata.

Conidiophores smooth- walled, more or less browned, usually sinuate,
commonly less than 200m long and terminating in dome-like or hemi-
spherical vesicles.

Conidia globose, echinulate, 3 to 4m in diame'ter.

Perithecia usually present; ascospores purple-red in color and character-
ized by equatorial bands.

Large, thick-walled, globose bodies, termed "hiille cells" (by Eidam),
forming an irregular layer around the perithecia.

Aspergillus (Sterigmatocyslis) nidulans was described by Eidam in 1883.
Since that time the general type of organism covered by his diagnosis has
become fairly well-known and certain striking characters have become
recognized as defining a number of cosmopolitan strains or species, com-
monly referred to as constituting the Aspergillus nidulans group.

The hiille cells of Eidam, first described in A. nidulans, appear also in

various transformations in the A. versicolor, A. ustus, and A . flavipes groups,

together with other common characters indicative of close relationship.

They do not appear in the A . clavatus, A . glaucus, or A . fumigatus groups

which are characterized by single sterigmata, nor do they occur in the

sclerotium forming groups, A. candidus, A. niger, A. wentii, A. tamarii, A.

flavus, and A. ochraceus.

Group Key
I. Ascospores present.

A. Ascospores smooth-walled.

1 . Equatorial ridges two in number.

a. Ridges 0.5 to 1.0m wide, margin entire.

Conidial heads green A. nidulans (Eidam) Wint.

Conidial heads white A. nidulans mut. alba Yuill

b. Ridges 1.5 to 1.8m wide, margin entire

A. nidulans var. latus Thom and Raper

c. Ridges 3.0 to 4.9m wide, margin dissected, starlike

A. variecolor (Berk, and Br.) Thom and Raper

2. Equatorial ridges usually four in number

A. quadrilineatus Thom and Raper

B. Ascospores rough-walled A. rugulosus Thom and Raper

1 Abridged from: Thom and Raper, The Aspergillus nidulans group, Mycologia,
Vol. XXXI, No. 6, 653-669, Nov.-Dec. 1939.

155

156 A MANUAL OF THE ASPERGILLI

II. Ascospores lacking.

A. Hiille cells forming irregular masses, suggestive of sclerotia

A. caespitosus Raper and Thorn

B. Hiille cells absent; heavy walled sterile hyphae present

A. unguis (Emile-Weil and Gaudin) Thorn and Raper

Aspergillus nidulans (Eidam) Wint. in Rab. Krypt.-Fl. I2: 62. 1884.

Synonyms : Sterigmatocystis nidulans Eidam in Cohn, Beitr. Biol. Pflan-
zen 3 : 392-41 1. pi. 20-22. 1883.
Diplostephanus nidulans (Eidam) Langeron, Compt. Rend.
Soc. Biol. Paris, 87: 343-345. 1922.

Colonies upon Czapek's solution agar plane, spreading broadly, dark
cress green (Ridgway, PI. XXXI) from abundant conidial heads during the
first two weeks (PI. IA, IVC, and fig. 41 A) ; perithecia developing fron the
center of the colony outward after the first few days, separately produced,
often abundant (PI. IVC) ; sectoring occasional; reverse of colony in varying
shades of purplish-red during the growing period, becoming very dark in
age. Heads short, columnar, ranging from 40 to 80m by 25 to 40m, com-
monly 60 to 70m by 30 to 35m (PI- IB and fig. 4 C); conidiophores com-
monly sinuous, with walls smooth, in shades of cinnamon brown (PI. IC),
ranging from 60 to 130m, commonly 75 to 100m in length, about 2.5 to 3m
near the foot, increasing to 3.5 to 5m below the hemispherical vesicle (fig.
39 A); vesicle 8 to 10m in diameter; sterigmata in two series, primary 5 to
6m by 2 to 3m and secondary 5 to 6m by 2 to 2.5m; conidia globose, rugulose,
3 to 3.5m in diameter, green in mass.

Perithecia developed separately within or upon the conidial layer (PI.
IA), globose, ranging from 100 to 175m in diameter, commonly 125 to 150m,
with outer layer a yellowish to cinnamon colored envelope of scattered
hyphae bearing hiille cells up to 25m in diameter; wall composed of one
layer of cells, dark reddish-purple; in ripening becoming a mass of 8-spored
asci which break down quickly leaving the ascospores free. Ascospores
purple-red, lenticular, smooth-walled with 2 equatorial crests (PI. ID and
fig. 43 A), spore bodies about 3.8 to 4.5m in length by 3.5 to 4m in breadth,
equatorial crests pleated with margin sinuous and entire ranging from 0.5
to 1m in width (Table 1).

Diagnosis based primarily upon culture XRRL Xo. 187 (Thom Xo.
4640.5) obtained from the Bainier collection in Paris. Other strains as-
signed to Eidam 's species included many isolations from American soil and
decaying vegetation, as well as cultures from European contributors.
Common.

The range of ascospore measurements found in a representative group
of cultures is shown in the accompanying table.

In assigning Eidam's species name to the members of this series it is
obvious that there are discrepancies. He described the perithecium as

THE ASPERGILLUS NIDULANS GROUP

157

Fig. 39. Conidial structure in the Aspergillus nidulans group, X 900: Au Typical
conidiai head of A. nidulans, XRRL Xo. 187; Ao, Diminutive head of the same strain;
B, Typical head of .4. caespitosus, XRRL Xo. 1929.

Fig. 40. Aspergillus nidulans. A, Conidial heads, X 370. Conidiophores arise
either from the substratum or from aerial hyphae. B, Hiille cells, X 740.

Fig. 41. Aspergillus nidulans group; different species growing upon Czapek's
solution agar at room temperature. A, A. nidulans NRRL No. 194, typical strain
producing very abundant dark green conidial heads. B, Naturally occurring muta-
tion characterized by white heads, isolated from the preceding strain by Edward
Yuill and described as A. nidulans mut. alba. C, A. rugulosus NRRL No. 207,
characterized by heavy perithecium production, an almost complete absence of
conidial structures, and a tendency of colonies to split in central areas as shown.
D, A. variecolor NRRL No. 1954, characterized by the production of abundant large
perithecia. E, A. unguis NRRL No. 216, characterized by an absence of perithecia
and hulle cells. F, A. caespitosus NRRL No. 1929, characterized by the production
of masses of hiille cells suggesting abortive perithecia.

158

THE ASPERGILLUS NIDULANS GROUP

159

having a firm almost sclerotioid wall, whereas the wall is found to contain
but one laj^er of cells. He figured the asci as few and scattered in a mycelial
matrix within which ascospore production occupied many weeks. One
isolated strain in our collection produces asci in this manner. For it, the
varietal name, A. nidulans var. latus, was proposed by Thorn and Raper
(1939) on account of very broad crests on the ascospore in contrast to the
usual types in A. nidulans which come much more closely to those indicated
in Eidam's figures.

TABLE 1
Ascospore variation in strains of Aspergillus nidulans

Culture number

NRRL 193..
NRRL 188. .
NRRL 189..
NRRL 194..
NRRL 195*.
NRRL 191 . .
NRRL 187..
NRRL 192..
NRRL 198. .
NRRL 199..

Overall dimension of spores

6.2-6.6
6.0-6.6
6.0-6.4
5.4-5.8
5.4-5.8
5.4-5.8
5.4-5.8
5.0-5.4
4.8-5.4
4.8-5.2

x 3. 6-3. 8M
x 3. 6-3. 9/x
x 3. 6-3. 8m
x 3. 6-3. 8m
x 3. 6-3. 8m
x 3. 6-3. 9m
x 3. 6-3. 8m
x 3. 6-3. 8m
x 3. 6-3. 8m
x 3. 6-3. 8m

Width of crests

1.0m ±

1.0m ±

1.0m ±

0.6-0. 8m

0.6-0. 8m

0.6-0. 8m

0.7-0. 8m

0.5-0. 6m

0.5-0. 6m

0.5m ±

Dimensions of spore bodies

4.2-4.6 x 3. 6-3. 8m
4.0-4.6 x 3. 6-3. 9m
4.0-4.4 x 3. 6-3. 8m
4.0-4.4 x 3. 6-3. 8m
4.0-4.4 x 3. 6-3. 8m
4.0-4.4 x 3. 6-3. 9m
3.9-4.3 x 3. 6-3. 8m
4.0-4.4 x 3. 6-3. 8m
3.8-4.4 x 3. 6-3. 8m
3.8-4.2 x 3.6-3. 8m

* White mutant from culture NRRL No. 194; described by Yuill as Aspergillus
nidulans mut. alba.

A. nidulans mut. alba Yuill, in Jour, of Botany (London)

175, pi. 618. 1939.

Colonies of the variety on Czapek's solution agar differ from the species
in the entire absence of green color (fig. 41 B). The ascospores have the
characters of the species.

Culture obtained by Yuill as a mutant from a normal green strain of A.
nidulans under investigation in his laboratory. Our record number is
NRRL 195; the normal and parent strain is NRRL 194.

Aspergillus nidulans var. latus Thorn and Raper. Mycologia 31 : 657. 1939.

Colonies on Czapek's solution agar differing from the species in colony
development characterized by a felt of predominantly sterile mycelium;
few conidial heads; fairly abundant perithecia developed in the mycelial felt
and each surrounded by a thick covering of hlille cells (fig. 42 C), very slowly
ripening and containing few and scattered asci in abundant sterile mycelium.
Ascospore bodies smooth-walled, purple-red, 3.8 to 4.5m by 3.5 to 4m, with
crests 1.5 to 1.8m in width.

Type culture NRRL No. 200 received from the Centraalbureau in 1909
and remaining constant in culture since that time. Its antecedent history
is not known.

160 A MANUAL OF THE ASPP^RGILLI

Aspergillus quadrilincatus Thorn and Raper. Mycologia 31: 6G0, figs. 2-

4. 1939.

Colonies on Czapek's solution agar spreading, plane or slightly wrinkled,
with tendency toward floccosity, central area gray with a definite purplish
tinge, and olive-green conidial areas toward the margin, occasionally as
sectors; perithecia developing separately but abundantly throughout the
colony; reverse purplish-red; heads short columnar, green, mostly 60 to 70/z
by 30 to 35m, occasionally larger or smaller; conidiophores sinuate, smooth-
walled, dull brownish in color, 50 to 75/i in length by 3.5 to 4.5m wide,
broadenng to 7.5 to 9/x at the hemispherical vesicular areas; primary
sterigmata 5 to 6/x by 2 to 3m, secondary sterigmata 5 to 7 m by 2 to 2.5m;
conidia globose, pale yellow-green, rugulose, 3 to 4m in diameter; perithecia
enveloped by hiille cells, light brownish in color, spherical, partially em-
bedded in the mycelial felt (fig. 42 D), about 125 to 150m in diameter includ-
ing the enveloping hiille-cell layer, with perithecial wall 1-cell layer in
thickness, ripening quickly and with ripe asci breaking down to leave the
ascopores free; ascospores purple-red, lenticular, with smooth wall, with
spore body 4 to 4.8m by 3.4 to 3.8m, and with two pleated equatorial crests
about 0.5m in width paralleled by a secondary narrower pair (fig. 43 B)
which are sometimes indistinct.

Type NRRL No. 201 (Thorn No. 4138.N8) from New Jersey soil and kept
in culture since 1916. Other strains examined include isolations from
Texas, Colorado, Louisiana, and Maryland.

Aspergillus rugulosus Thom and Raper, Mycologia 31: 660-663,

fig. 4. 1939.

Colonies on Czapek's solution agar slowly and restricted^ growing (fig.
41 C), buckled or wrinkled in a mass 2 to 3 mm. deep, enveloping abundant
perithecia at different depths, often eventually splitting in the central area,
purple-gray to purple-brown in age, with green heads sparsely produced
and hence not generally evident, occasionally seen as small groups and
marginal extensions into drying media; reverse in shades of deep purple-
red; conidial heads, short columnar, 75 to 100m by 30 to 40m; conidiophores
sinuous, smooth-walled, pale brownish in color, 50 to 80m long, slender,
varying up to 5m in width, then enlarging to vesicular hemispheres 8 to 10m
in diameter; primary sterigmata 7 to 8m by 3 to 3.5m, secondary sterigmata
6 to 7m by 2.5 to 3m; conidia globose, green, rugulose, 3 to 4m.

Perithecia very abundant, often imbedded in the mycelium as 2 or 3 layers
and each surrounded by hyphae and dark brown hiille cells (fig. 42 E),
globose, 225 to 350m in diameter including mycelial coverings, with dark
reddish-purple walls of one cell thickness, quickly ripening and breaking
down to leave ascospores free; asci 10 to 11m in long axis; ascospores purple-
red, lenticular, walls conspicuously rugulose (fig. 43 C), with spore bodies 4

THE ASPERGILLUS NIDULANS GROUP

161

to 4A/J. by 3.6 to 3.8ju, and with 2 pleated equatorial crests with sinuate and
entire margins about 0.5 to 0.6m in width.

A: A. nidulans

B: A. unguis

immBm

O A. nidulons vor. lotus

D: A. quodrilmeotus

IS&&>»S8J&&_suk s&sg^^Ssfc^sisi&s

E ■' A. rugulosus

F: A. voriecolor

Fig. 42. Diagrammatic representations of cross sections of colonies of different
species of the Aspergillus nidulans group showing the relative abundance of conidial
heads and perithecia and the manner in which these structures are borne : con, conidial
heads; d.h., mantle of divergent hyphae; hul, huile cells; my, mycelial felt; per,
perithecia; ps, pseudostalk of hiille cells and sterile hyphae; st, long, thick walled,
sterile hyphae; sub, substratum; and un, unstalked perithecium. Scale approxi-
mate. (Reprinted from Thom and Raper, "The Aspergillus nidulans Group,"
Mycologia 31: 653-669. 1939.)

Cultures studied included Type NRRL No. 206 (Thom No. 4138.T11)
from New Jersey soil, as discussed by Thom and Church in The Aspergilli,
p. 138, and also isolates from Washington, D. C, Texas, Nebraska, and
California. Very common in soil.

162

A MANUAL OF THE ASPERGILLI

Fig 43 Ascospores of different species of the Aspergillus nidulans group. A, A.
nidulans. B,A.quadrilineatus. C,A.rugulosus. D, A. vanecolor In each species
upper left and right and center left spores represent surface profile views; center
right, surface in face view; lower left, optical section in profile; and lower right,
optical section in face view. (Reprinted from Thorn and Raper, 'The Aspergillus
nidulans Group," Mycologia 31: 653-669. 1939.)

THE ASPERGILLUS NIDULANS GROUP 163

Culturally and microscopically the above strains present similar pictures,
with the exception of the strain recently received from Bliss in California
(NRRL No. 2 1 1 ) . In contrast to the others, this culture produces abundant
conidial heads and relatively fewer perithecia. The ascospores and conidial
structures, however, duplicate those of the typical strains; hence we do not
at present feel warranted in designating this as a variety, or otherwise
separating it from the species A. rugulosus.

Aspergillus variecolor (Berk, and Br.) Thorn and Raper, in Mycologia 31:

663-G67. fig. 4D and fig. 5. 1939.

Synonyms : Emericella variecolor Berk, and Br. in Berkeley, Introd.

Crypt, Bot. p. 340-341; fig. 76. 1857. See Patouillard,

Bull. Soc. Myc. Fr. 7: 43-49. pi. 4. fig. 6-12. 1891.
Inzengaea erythrospora Borzi, Jahrb. Wiss. Bot. (Pringsheim)

16: 450-463. pi. 19, 20. (1884) 1885.
Emericella medias Chowdhury and Mathur, Ann. Myc. 36:

61-63. 1938.
Aspergillus stellatus Curzi, Rend. Acad. Naz. Lincei 19:

424-428. fig. 1. 1934.

Colonies on Czapek's solution agar with vegetative mycelium largely
submerged, sparse, spreading slowly in the agar, producing green heads
freely in the center of the colony, less abundantly in the outer areas, large
gray perithecia produced in clusters in colony center and at the margin in
some strains, with smaller perithecia scattered through the intervening
thinner areas of the colony (fig. 44 A), in other strains producing large
perithecia abundantly throughout the colony (PI. IVD); reverse color in
shades of purple-red. Conidial heads green, columnar (fig. 44 D), relatively
long, mostly 100 to 200m, occasionally up to 300m by 30 to 40m; conidiophores
arising directly from submerged hyphae, straight with smooth walls, cinna-
mon-brown in color, mostly 140 to 200m long by 3 to 5m in diameter,
broadening gradually to become hemispherical vesicles about 8 to 10m in
diameter; primary sterigmata 7 to 8m by 3 to 4m, secondary sterigmata 8 to
9m by 2.5 to 3m; conidia globose, rugulose, 3 to 3.5m; perithecia when clus-
tered (fig. 44 Aa) 300 to 400m in diameter surrounded by a felt of hyphae
and hulle cells and supported by masses of hyphae and hulle cells forming
false stalks (fig. 42 F), giving the structures a pyriform appearance (fig.
44 B) ; scattered perithecia much smaller (fig. 44 C) and with envelope of
supporting cells often much reduced in mass; hulle cells abundant and
essentially like those of the species A. nidvlans.

Perithecial wall when stripped of enveloping cells purple-red, brittle,
composed of a single layer of cells; asci quickly ripening and breaking down
to leave the cavity filled with ascospores; ascospores purple-red, with spore
bodies lenticular and 3.6 to 4m by 2.8 to 3m, with two prominent equatorial

164

A MANUAL OF THE ASPERGILL]

Fig. 44. Aspergillus variecolor. A, Central area of colony; a, cluster of large
pseudostalked perithelia; b, scattered, smaller, unstalked perithecia (see also figure
42 F); c, scattered conidial heads, X 6. B, Enlarged view of large, pseudostalked
perithecium, X 65. C, Enlarged view of small, unstalked perithecia, X 65. D,
Enlarged view of conidial heads, X 65. (Reprinted from Thorn and Raper, "The
Aspergillus nidulans Group," Mycologia 31: 653-669. 1939.)

crests, up to 3.5m in width, pleated and cot to give a stellate appearance to
the ascospores (fig. 43 D).

THE ASPERGILLUS NIDULANS GROUP 165

The above description is based primarily upon a culture received from
Prof. Verona in Italy and carried in our collection as NRRL No. 212 (Thorn
No. 5602.3).

Bliss forwarded a culture (NRRL No. 214) isolated from date fruits in
California which differs from the above in the following particulars: (1)
Conidial heads are produced abundantly, (2) the mycelium is not predomi-
nantly submerged, and (3) the colonies in reverse are deep purple. How-
ever, the ascospores of the two strains are strikingly similar in size and
pattern, the perithecia of each appear pyriform in shape, and the conidial
structures of the two are essentially alike. More recently a strain has been
isolated from Arizona soil (NRRL No. 1954) which produces abundant large
"stalked" perithecia but very few small perithecia or conidial heads.
There is, thus, evidence of considerable natural variation among members of
this species. All of these strains have the same type of stellate ascospore.

Since the type of ascospore described here had already been assigned to
Emericella and Inzengaea, consideration of the literature of these genera is
necessary.

Emericella variecolor, genus and species new, was described by Berkeley
and Broome in 1357 as doubtfully a Gasteromycete or possibly a lichen.
The perithecium with a mass of stellate spores was considered as gastro-
mycetous in character while what we now recognize as the "hulle" cells of
Eidam (figured) suggested to them the possibility of an algal associate.
Berkeley's material was also examined by Montague and part of it deposited
in the Museum d'Histoire Naturelle de Paris. This was reexamined by
Patouillard in 1891 and its ascomycetous nature determined. No conidial
apparatus was found by either Berkeley or Patouillard.

Inzengaea erythrospora as the type species of a new ascomycetous genus
was figured and described by Borzi in 1885, showing stellate red ascospores,
and the hulle cells of Eidam. Borzi 's figure showed a coremium-like conid-
ial apparatus which was designated Coremium Borzianum by Saccardo.
Ed. Fisher in 1893 transferred the species to Emericella of Berkeley and
placed the genus next to Aspergillus in the "Pflanzenjamilien." Saccardo
(in Syll. 9: 610) on the other hand accepted Inzengaea and dropped Emeri-
cella because of the errors in description and placement by Berkeley. Borzi
figured the spores of A. variecolor {Inzengaea erythrospora) correctly but
obviously misinterpreted the germination of the ascospores since he showed
them splitting as if turned 90°, bringing the crest perpendicular to the
center of the valve instead of attached to its edges.

There the taxonomic situation stood until Vuillemin in 1927 concluded
that the ascosporic apparatus, the stellate red ascospores and the cells of
Eidam, as clearly shown in their figures and material, showed the identity

166 A MANUAL OF THE ASPERGILLI

of Emericella and A. nidulans. He therefore transferred A. nidulans to
Emericella as the oldest established genus and apparently did not even
consider Inzengaea.

Ciferri (1938) has recently completed a study of Emericella variecolor
embracing cultural investigations together with a review of the literature
of the genus. He did not recognize the close relationship of this fungus to
Aspergillus nidulans, and was apparently unmindful of the likeness of their
conidial structures and the essential similarity of their perithecia and asco-

spores.

In 1934 Curzi described as Aspergillus stellatus a fungus characterized by
hulle cells and red, stellate ascospores. However, he apparently did not
know of either Berkeley's or Borzi's earlier designation of a similar fungus.
It is unfortunate that his exceedingly descriptive binomial must be reduced
to synonymy.

Fortunately for this discussion, cultures NRRL Nos. 212, 214, and 1954
present both the stellate spores and apparently stalked perithecia figured
by Berkeley, Patouillard, and Borzi. (Compare figures 42, F, and 44,
B with Berkeley's figure 76a, Patouillard 's figures 7 and 8, plate 4, and Bor-
zi's figure 10, plate 19.). This made possible a restudy of the whole
morphologic situation from fresh material. The perithecial body itself
was found not to be stalked but to rest upon a sterile mass of hulle cells
and mycelium giving the superficial appearance noted by earlier observers.

The coremium of Borzi remains unaccounted for. Obviously in Borzi's
discussion the material was rotten olives and very old. No cultures were
made. The conidia-producing apparatus (figured) differs essentially in
type from the conidial apparatus of the Aspergillaceae with which Fischer
correctly placed Emericella because of its perithecia and ascospores. We
are convinced that the coremia belonged to some other fungus.

It is not possible to separate the perithecium of this fungus from that of
the other species in the A. nidulans group nor are there characters to take
this type of perithecium out of a genus with the yellow perithecium of the
great A . glaucus series of species which are widely known. Consistent with
the policy of keeping the Aspergilli in one group, both Emericella and In-
zengaea are dropped for purposes of this discussion.

Aspergillus caespitosus Raper and Thorn, in Mycologia 36: 563-565,

fig. 4. 1944.

Colonies varying markedly upon different media; upon Czapek's solution
agar rather slow growing, attaining a diameter of 6 to 8 cm. in three weeks
at room temperature, plane or somewhat furrowed, mycelium largely sub-
merged and extremely tough, tearing with difficulty, producing numerous
dark green, hemispherical to loosely columnar heads in central colony areas,

THE ASPERGILLUS NIDULANS GROUP

167

characterized particularly by clusters of irregular ovoid to elliptical, thick-
walled hulle cells, at first colorless becoming reddish-purple in age, scattered

**.

8

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V

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Fig. 45. Aspergillus caespitosus ; , NRRL No. 1929. A, Portion of colony on
Czapek's solution with 1 percent liver extract, showing crowded conidial heads in
central portion and scattered hulle cell masses in surrounding areas, two weeks old,
incubation at room temperature, X 1.8. B, Portion of colony enlarged showing
hulle cell masses, X 10. C, Silhouettes of conidial heads developed on hay infusion
agar, X 48. D, Typical conidial head showing form of vesicle and arrangement of
sterigmata, X 600. E, Portion of htille cell mass showing irregular size and form
of component elements, X 265. (Reprinted from Raper and Thorn, "New Species
of Aspergilli from Soil," Mycologia 36: Nov.-Dec. 1944.)

unevenly (fig. 45 A and B) or arranged in irregular concentric zones; reverse
colorless at first, becoming dark reddish-purple in age, particularly beneath
the hulle masses; odor none. Conidial heads dark dull yellow-green to

168 A MANUAL OF THE ASPERGILLI

empire green (Ridgway, PI. XXXII), generally hemispherical to loosely
columnar, mostly 75 to 125m in diameter. Conidiophores straight or
slightly sinuous (fig. 45 D), mostly 250 to 325m in length, occasionally up
to 350m by 5.0 to 6.5m in diameter, of approximately uniform diameter
throughout, relatively thick-walled, (1.2 to 1.5m in basal portion to 0.8
to 1.0m in terminal area), smooth, tan to light brown in color. Vesicle
slightly elongate, the upper hemisphere loosely covered by sterigmata
(fig. 45 D), the lower half sterile and often lightly colored, mostly 15 to 20m
in diameter. Sterigmata in two series (fig. 45 D), primaries normally 6.5
to 8.5m by 3.5 to 5.0m, secondaries 6.5 to 8.0m by 3.0 to 4.5m, typically bottle
form but commonly much swollen and often quite irregular in form and
dimensions. Conidia globose, spinulose, green, mostly 3.5 to 4.5m, rarely
larger, hiiHe cells very abundant, thick-walled, irregularly globose, ovoid or
elliptical (fig. 45 E), ranging from 12 to 18m in globose cells to 12 to 15m
by 25 to 30m in the most elongate bodies, forming compacted masses of
indefinite size, extremely tough and in age becoming almost sclerotioid, at
first colorless but in age characterized by an abundant reddish-purple
intercellular pigmentation.

Colonies upon malt agar characterized by a dense stand of erect conidio-
phores bearing hemispherical to radiate or loosely columnar heads of dark
green color approximately empire green (Ridgway, PL XXXII) and the
complete absence of hulle cells; reverse in light brown shades; odor none.
Details of morphology as upon Czapek's solution agar.

Colonies upon hay infusion agar like those upon malt except less heavily
sporing.

Strains include NRRL No. 1929 (type) isolated from Arkansas soil and
other isolations from Arizona and Texas soils.

This species is of particular interest because of its apparent transitional
position between the A. nidulans group and A. ustus. In the character
of its conidiophores, its reddish-purple pigmentation, and in the general
color and markings of its conidia it retains the characters of A. nidulans
and closely related species. In the absence of fertile perithecia and asco-
spores, the predominantly hemispherical shape of its conidial heads, and in
the variable and irregular form of its hiiHe cells, it is strongly suggestive of
the A. ustus series. While we are convinced of its. intermediate position
between the A. nidulans group and A. ustus, we place it with the former
since we believe it is most closely allied to this group. It is believed sig-
nificant that superficially, cultures of Aspergillus caespitosus and Aspergillus
variecolor (Berk, and Br.) Thorn and Raper (1939) are strikingly similar
upon Czapek's solution agar. This similarity is particularly marked when
plates are viewed in reverse since an intense pigmentation marks the under
surface of perithecia in the latter case and the under surface of older hulle
masses in the former.

THE ASPERGILLUS NIDULANS GROUP

169

Aspergillus unguis (Emile-Weil and Gaudin) Emend. Thorn and Raper,

Myc. 31, p. 667, fig. 6. 1939.

Synonyms: Sterigmatocystis unguis Emile-Weil and Gaudin, Arch. Med.
Expt. Anal. Path. Paris 28: 463-465, fig. 4, 1919.
A. loakiashanensis Shih, Lingnan Sci. Jour. 15 (3): 369. p.
16, fig. 2. 1936.

Colonies on Czapek's solution agar restrictedly growing, plane, spreading
at the margin as irregular lobes (fig. 41 E), yellowish-green, green to dark
green becoming brown in age; without perithecia or hulle cells. Mycelial
preparations show striking sterile, thick-walled hyphae with walls in brown

Fig. -46. Sterile spicule hyphae of Aspergillus unguis. A, Cluster of sterile
hyphae, X 370. B, Apex of sterile hypha, X 740. C and D, Mid-portions of sterile
hyphae showing thick roughened walls, X 740. (Reprinted from Thorn and Raper,
"The Aspergillus nidulans Group," Alycologia 31: 653-669. 1939.)

shades, irregularly roughened (fig. 46), tapering to a blunt point, arising
sometimes from foot-cells suggesting the origin of conidiophores, sometimes
apparently from mycelial cells, often up to 1,000m or more in length, slant-
ing upward but usually rising only slightly above the conidial area (fig.
42 B).

Conidial heads columnar, 75 to 150m by 40 to 50m; conidiophores smooth-
walled, dull brown in color, mostly 45 to 65m in length by 3 to 5m in diameter,
enlarging to vesicular hemispheres 9 to 12m in diameter; primary sterigmata
5 to 6m by 2.5 to 3m, secondary sterigmata 5 to 6m by 2 to 2.5m; conidia
globose, rugulose, dull green, 2.5 to 3.5m in diameter.

Cultures of A. unguis are obtained frequently from medical laboratoies
apparently as more or less active pathogens but occasionally isolated from

170 A MANUAL OF THE ASPERGILLI

soil and decaying organic matter. The question whether the non-
ascosporic members of the group have merely dropped the ascogenous
phase or constitute a separate species was answered when more complete
examination showed the sterile or spicule hyphae to be regularly produced
in the non-ascosporic, but never found in ascosporic series.

Pathogenicity

Aspergillus nidulans in some of its forms and variants has been demon-
strated as a parasite in human nails (onychomycosis), often enough to
establish its pathogenicity. A. nantae Pinoy (1927) probably belongs
here although the data are mainly pathological, hence not adequate for
definite identification of the organism. A. nidulans forme cesarii Pinoy
(1915) isolated from a mycetoma of the lung of a donkey, and A. nidulans
var. nicollei Pinoy (1906) isolated in Tunis from a case of mycetoma, or
madura foot, represent additional strains which were at least secondary
pathogens. The nearly related A. unguis is usually the more common
form isolated from human material. A. Brodeni (Mattlet) Dodge (1935)
from a bronchomycosis in Africa might have been close to A. unguis. A.
nidulans and A. unguis are both widely distributed as saprophytes; hence
are constantly encountered as components of dirt reaching the extremities
by contamination. Infection of the air passages is comparatively rare.

Occurrence and Economic Importance

In addition to their role as occasional disease producing agents, members
of the A. nidulans group are believed to be significant in decomposition
processes. They are among the molds most commonly isolated from soil,
and very frequently appear in considerable abundance upon vegetable ma-
terial undergoing slow decomposition. Aspergillus rugulosus, A. quadriline-
atus, and A . caespitosus occur most frequently in soils from the comparatively
dry, warm soil of Texas, Arizona, and adjoining areas. Aspergillus varie-
color has been isolated from olives in Italy, date fruit in California, and from
Arizona soil. Aspergillus nidulans is abundant and cosmopolitan in its
distribution.

Chapter XII
THE ASPERGILLUS USTUS GROUP

Outstanding Characters

Colonies more or less floccose, at first white but becoming dull in age, in
most members varying from olive-gray through reddish-brown to
fuscous, as conidial structures develop.

Conidiophores in yellow-brown shades; smooth.

Heads irregular in form, ranging from more or less radiate to hemi-
spherical to loosely columnar.

Vesicles hemispherical; sterigmata in two series, loosely arranged.

Conidia roughened, 3.0 to 5.0m, varying from echinulate to marked with
conspicuous color bars, and ranging in color from pale blue-green
through olive-green shades to deep brown (fuligineus).

Hulle cells regularly present, thick-walled, elongate, often more or less
curved and twisted.

Included here are representatives of a most abundant and widespread
group of fungi, especially common in soil and upon decaying vegetation.

Group Key

Colonies predominantly floccose, heavy sporing, hiille cells not aggregated in small

clusters A. ustus (Bain.) Thorn and Church

Colonies more or less floccose, light sporing, hiille cells aggregated in small clusters

A. granulosus Raper and Thorn

Aspergillus ustus (Bainier) Thorn and Church, in The
Aspergilli, p. 152. 1926.

Synonym : Sterigmatocystis usta Bainier, in Bui. Soc. Bot. France 28 :
78. 1881.

Colonies upon Czapek's solution agar spreading broadly, plane, sulcate,
or umbonate, rarely zonate, more or less felted or floccose; at first white,
becoming olive-gray, yellow-brown, fuscous or russet to purplish vinaceous
with the development of mature conidial structures (PI. IV E, and figs.
48 A and B) ; generally heavy sporing, with some conidiophores arising from
the substratum but more abundantly from aerial hyphae ; reverse in shades
of yellow, orange, and brown to almost black in age; odor not pronounced.
Heads radiate to irregularly hemispherical, sometimes loosely columnar,
commonly splitting into more or less well-defined columns in age, very
variable in size, ranging in color from dull green or olive-gray, through gray-

171

172

A MANUAL OF THE ASPERGILLI

ish-brown to fuscous or fuligineus. Conidiophores arising from submerged
hyphae (fig. 47 A) ranging up to 500m long by 3 to 6m, aerially borne conidio-
phores, ranging from very short (fig. 47 Ai) up to 125m by 2 to 5m, sinuous,
sparsely septate, with walls rather thin, smooth, and uniformly colored
some shade of brown. Vesicles hemispherical to subglobose, 8 to 20m in
diameter, smaller in some strains (fig. 47 A). Sterigmata colorless or

Fig. 47. Aspergillus ustus group, X 840. A, Typical conidial head showing com-
paratively loose sterigmata in two series and conspicuously roughened conidia, strain
NRRL No. 278. Au Diminutive head, as often seen in strain NRRL No. 275. B,
Typical head of Aspergillus granulosus, NRRL No. 1932.

colored, semi-radiate, loosely arranged into two series, primary sterigmata
4 to 7m by 3m, secondary sterigmata 5 to 7m by 2.0 to 2.5m- Conidia globose,
3.5 to 5.0m, roughened, echinulate to marked with conspicuous color bars,
ranging from greenish through olive-gray to yellow-brown or fuligineus.
Many strains producing thick-walled hulle cells (fig. 49 E) ranging in form
from irregularly ovate or elongate in some strains, to serpentine, helicoid,
or twisted in others, essentially as in Aspergillus flavipes.

THE ASPERGILLUS USTUS GROUP

173

Upon malt agar, colonies frequentl,y heavier sporing and conidial heads
generally tending to run to dull gray-green shades rather than brown.

Very common in soil and decaying vegetation. Species diagnosis repre-
sents a composite based upon many isolations from this country and abroad.

B

D

Fig. 48. Aspergillus ustus: cultures growing upon Czapek's solution agar at room
temperature, 10 days. A, Strain XRRL No. 275 characterized by loose floccose
colonies and moderate sporulation. B, Strain NRRL No. 278 characterized by
heavier spore production and the presence of abundant hiille cells. C, A. ustus var.
laevis, NRRL No. 1852, characterized by loose floccose colonies and conidial heads
often near brick red in color. D, Strain NRRL No. 1974 characterized by the pro-
duction of very abundant hiille cells in concentric zones.

Individual strains differ markedly in their general habit and colony colora-
tion, in the color of their fruiting structures, in the marking and coloration
of their conidia, in the presence or absence of hiille cells, and in the form of
these structures when present. By a deliberate selection of strains, one
can find sufficient difference to warrant the assignment of specific designa-

174 A MANUAL OF THE ASPERGILLI

tions to particular cultures. Yet all possess the characters noted above and
so constitute a well-defined group whose variations are matters of detail.
Such differences as occur, moreover, tend to become bridged as comparative
cultural and microscopic studies of many isolations are made ; hence the use
of these differences becomes of questionable value for diagnostic purpose.
We have considered it desirable, therefore, to include the whole series under
one name as a single species aggregate, Aspergillus ustus, and to call atten-
tion to some of the major differences which one may expect to encounter
among the members of this species. Although Bainier was not sufficiently
explicit in his description of Sterigmatocystis usta to enable us to identify
with certainty the form with which he worked, his usage is accepted upon
(1) the basis of priority, and (2) the receipt of a culture (Thorn No. 4640.
488) from his laboratory labeled Sterigmatocystis usta, which possessed the
basic characters of the group as herewith set forth.

Long after the publication of Sterigmatocystis usta, Bainier described a
second species belonging to this group, Sterigmatocystis insueta (Bui. Soc.
Mycol. France 24: 85-87, PL VIII, Fig. 1-13. 1908), and emphasized
the fuligineus character of its colonies, the predominant origin of brown
fruiting structures from aerial mycelium, the larger size and the darker
color of its conidia, which were characterized by the presence of pronounced
color bars. Strains showing these characters probably represent the type
most commonly encountered among miscellaneous isolations of forms be-
longing to this group. Recognition of these forms as constituting a distinct
species has been considered, but in the absence of any clearly definable line
of separation from A. ustus it is believed desirable to leave them within the
somewhat extended framework of this species. Cultures possessing these
characteristics very commonly exhibit hiille cells varying in different strains
from ovoid to irregularly elongate, to serpentine, helicoid or otherwise
twisted. One strain showing much-twisted hiille cells was isolated and
contributed by Thaxter under the manuscript name A. helicophorus.
Typically, the conidiophores of these forms are grayish-brown, commonly
quite dark. The vesicle and sterigmata are likewise frequently colored.
Conidial color as seen under high magnifications varies with age from pale
to olive green to fuligineus, and conidial markings from fairly coarse
echinulations to intensively colored bars and tubercles.

Possibly unaware of the existence of Sterigmatocystis usta and S. insueta
(since no mention is made of either species), Abbott (1926) subsequently
described Aspergillus minutus. His description indicated that he was
dealing with a strain essentially similar to those considered by Bainier as
S. insueta. This is confirmed by examination of his type culture, NRRL
No. 283 (Thorn No. 4894.2). A second culture, NRRL No. 285 (Thorn
No. 4894.1), received from Abbott under the manuscript name A. humus
likewise represents a member of this series and differs from the more

THE ASPERGILLUS USTUS GROUP 175

common forms only in its more floccose habits and in the production of
somewhat smaller conidial heads.

Among the more striking members of the Aspergillus ustus series
examined are two isolations from soil collected in Panama and Mexico,
respectively, which upon Czapek's solution agar normally produce strongly
zonate colonies consisting of alternating areas of crowded conidial heads
and heavy hiiHe cell development (fig. 48 D). Except for this difference in
colony appearance, however, these strains appear to be typical of the species
as described above.

A single strain, NRRL No. 1852, isolated from Louisiana soil, possesses
conidial heads of a dull brick red color (Ridgway, PI. XXXIX: russet
vinaceous to sorghum brown) and abundant, much-twisted hulle cells.
While Blochwitz's description is too inadequate to permit of detailed com-
parison, it is suggested that this strain may represent his Aspergillus ustus
var. laevis (Ann. Mycol. 32(1/2): 4. 1934), which was described as
characterized by "red conidia and crooked hiiHe cells." In strain NRRL
No. 1852, the conidia are conspicuously reddish en masse but appear only
slightly colored when viewed with high magnifications. In contrast to
most strains of the Aspergillus ustus group, the conidia are finely echinulate
rather than coarsely roughened. It is suggested that this form (fig. 48 C)
may represent a transition in the direction of Aspergillus flavipes since the
latter species is likewise characterized by much-twisted hiiHe cells, brown
conidiophore walls, and conidia which may show a reddish color in some
strains but are smooth in all.

Aspergillus granulosus Raper and Thorn, in Mycologia 36: 565-568,

fig. 4. 1944.

Colonies upon Czapek's solution agar growing well, attaining a diameter
of 8 to 10 cm. in two to three weeks at room temperature, plane or irregu-
larly furrowed, predominantly floccose, uneven in texture, buff to dull brown
in color from felted sterile mycelia ; conidial heads few in number and gen-
erally arising from the substratum direct, less often from aerial hyphae,
commonly appearing in clusters, pale blue-green in color; colonies charac-
terized particularly by abundant small, colorless clusters of irregularly
globose, ovoid, or elliptical thick-walled hiiHe cells which superficially sug-
gest perithecial initials and which in mass give to the colony a semi-granular
appearance (fig. 50 B, E, and F); reverse in shades of dull yellow and
brown; slight mushroom odor. Conidial heads few in number, commonly
clustered in small groups, most abundant at colony margin, sometimes
occurring on tufts of aerial hyphae, hemispherical to radiate, 75 to 125/x
in diameter, very loose, consisting of comparatively few divergent spore
chains (fig. 50 C), approximately pale niagara green in color (Ridgway,
PI. XXXIII). Conidiophores erect, straight, nonseptate, mostly 350 to

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Fig. 49. Htille cells. A, Characteristic thick-walled, globose to subglobose hiille
cells of the Aspergillus nidulans group, A. variecolor, NRRL. No. 1954, X 450. B,
Aspergillus caespitosus, strain NRRL No. 1930, hiille cells irregular in form and
often poorly developed but of the same general pattern as A. nidulans, X 275. C,
Aspergillus janus, NRRL No. 1787, hiille cells approximately globose and of the
same basic type as in A. nidulans, X 450. D, Aspergillus granulosus, NRRL No.
1932, hiille cells globose to ovoid, somewhat intermediate between A. nidulans and
A. ustus, X 275. E, Aspergillus ustus, NRRL No. 280, characteristic hiille cells,
elongate and much twisted, X 450. Hiille cells in Aspergillus flnvipes are of the
same pattern. F, Aspergillus carneus, NRRL No. 1926, heavy walled, strongly
septate mycelium suggestive of hiille cells, X 275.

176

THE ASPERGILLUS USTUS GKOUP

177

500^ in length, 5.5 to 8.0/x in diameter, approximately uniform in width
throughout, thin-walled, smooth, tan to light brown in color, often slightly

D

Fig. 50. Aspergillus granulosus; NRRL No. 1932. A, Portion of colony on hay
infusion agar showing clusters of conidial structures and small masses of hiille cells,
two weeks old, incubation at room temperature, X 1.5. B, Portion of marginal area
of two colonies on malt extract agar showing characteristic granular appearance
resulting from numerous small clusters of hiille cells, X 1.5. C, Silhouettes
of conidial heads, X 48. D, Conidial head showing slightly elongate vesicle, sterig-
mata in two series, and constriction in conidiophore just beneath the vesicle, X 750.
E, Small clusters, or "granules", of hiille cells, X 48. F, Double cluster of hiille
cells, much enlarged, X 265. (Reprinted from Raper and Thorn, "New Species of
Aspergilli from Soil," Mycologia 36: Nov .-Dec. 1944.)

constricted just beneath the vesicle (figs. 47 B and 50 D). Vesicle ovate
to elliptical, thin-walled and easily broken, largely covered by sterigmata,
12 to 18^ m diameter by 15 to 25/z in length. Sterigmata in two series,

178 A MANUAL OF THE ASPERGILLI

both comparatively short and stout; primaries 3.5 to 5.0m by 3.0 to 4.0m;
secondaries 4.0 to 5.5m by 3.0 to 3.5/z, commonly bottle-form. Conidia
globose, pale green, delicately echinulate. Mostly 4.8 to 5.5/x in diameter,
rarely larger. Hiille cells abundant, irregularly globose, ovoid or somewhat
elongate, commonly 12 to 30m in long axis, walls heavy, 4 to 5m in thickness,
borne primarily in small, colorless clusters which are quite conspicuous at
colony margins and lend to them a characteristic granular appearance.

Colonies upon malt extract agar showing an accentuation of hiille cell
development (fig. 50 B) and a reduction in conidial heads. Otherwise
duplicating the cultural picture presented upon Czapek's solution agar.

Colonies upon hay infusion agar (fig. 50 A) thin but broadly spreading,
characterized by scattered clusters of hiille cells and erect conidial fructifica-
tions giving to the culture a sparsely granular appearance.

Type culture NRRL No. 1932 was isolated in 1942 from a sample of soil
collected in Fayetteville, Arkansas, and contributed by Mr. F. R. Earle.
Additional strains have been isolated from soils collected in Texas, Arizona,
and Costa Rica. It is believed common in soils where the temperature
remains at a high level during part or all of the year.

Different strains vary materially in the number of conidial heads pro-
duced upon common laboratory media such as Czapek's solution and malt
extract agars, ranging from abundant heads in some to only widely scattered
heads in others. All fruit reasonably well, however, upon hay infusion agar,
the medium upon which original isolations were made.

The brown color of the conidiophores, the presence of ovoid to somewhat
irregular hiille cells, and the green color of its conidia place this species in
the group with A. ustus. It differs markedly from the more common
representatives of this group, however, in the lighter and persistently green
color of its conidia, the small clusters rather than irregular masses of hiille
cells, and in possessing somewhat more elongate vesicles. In this latter
character it suggests Aspergillus flavipes but is in turn excluded from this
group by the green color of its spores.

Occurrence and Economic Importance

Representatives of the Aspergillus ustus group are perhaps the most
abundant of all aspergilli in soil. They regularly occur in large numbers
and in considerable variety. The common species, A. ustus, occurs alike
in cultivated and forest soils and in approximately equal abundance in
soils from southern and from north temperate areas. A. granulosus, on
the contrary, has been isolated only from southern sources. Members of
the group are not known to be particularly active agents of decomposition,
but their great abundance in nature is believed indicative of a significant
role in many decay processes.

Their biochemical activities and potentialities are almost completely
unknown.

Chapter XIII
THE ASPERGILLUS FLAVIPES GROUP

Outstanding Characters

Conidiophores smooth, in some shade of yellow, with color often confined

to the outer layer.
Heads barrel-form to columnar when well developed, white or slowly

becoming some shade of vinaceous-buff to avellaneous (Ridgway,

PL XL).
Vesicles subglobose to elliptical.
Conidia colorless, smooth, thin-walled.
HiiHe cells generally present, helicoid or variously twisted.

The range of variation presented within the group has led workers with
only a few representatives before them to offer names and descriptions for
the strains under observation. However, when large numbers of strains
are brought together and cultivated upon a considerable range of substrata
the continuity of the group as a natural and related series of strains becomes
apparent. Further study of this whole group may lead to separation upon
lines accounting for certain published descriptions not at present identifi-
able. For purposes of the present manual, however, it is believed desirable
to broaden the description of Aspergillus flavipes (Bain, and Sart.) Thorn
and Church sufficiently to include a closely related series of organisms
rather than restrict it to the particular strains studied by Bainier.

Aspergillus flavipes (Bain, and Sart.) Thorn and Church,
The Aspergilli, p. 155. 1926.

Synonym: S. flavipes Bainier and Sartory (Bui. Soc. Myc. France 27:
90-96, PL HI., fig. 1-6. 1911).

Colonies upon Czapek's solution agar rather slow growing, becoming 3
to 5 cm. in diameter in about 10 days; mycelium yellowish, dull buff, com-
monly becoming brownish in age; heads pale to dull buff in some strains
to avellaneous or even very light cinnamon in others (PL IV F and fig.
51 A), submerged mycelium persistently colorless in some strains, develop-
ing many shades of yellow, orange, orange-brown to red (Madder-brown
of Ridgway, PL XIII) or almost black in reverse of colonies in others;
in some strains producing at the surface of the agar closely woven yellow
to orange masses of hyphae enmeshing numerous helicoid or variously
twisted, thick-walled hiille cells; occasional strains showing dark masses

179

ISO

A MANUAL OF THE ASPERGILLI

suggestive of sclerotia; aerial mycelium more or less abundant, colorless
or yellow to orange; producing in many strains numerous large drops of
transpired fluid, pale to yellow or orange-red (acting as an indicator, chang-
ing from yellow with acid to orange-red with alkali) ; commonly

Fig. 51. Aspergillus Jlavipes group. A,
Czapek's solution agar. 2 weeks, room tei

.4. flaripes XRRL No. 295 growing on
temperature. B, Strain XRRL No. 287 of
the same species, found bv White (1943) to produce a penicillin-like substance. C,
Strain XRRL Xo. 1959 characterized by the production of an excessive amount of
exudate. D, Photomicrograph of a typical head showing the elongate vesicle and
crowded sterigmata in two series, X "50.

characterized by a disagreeable odor approaching putridity. Heads
typically becoming columnar masses, shading from persistently white,
through shades of pale to deep avellaneous as in A. terreus; but usually in
rather sharp contrast to the color of the mycelium. Conidiophores from
300 to 500m by 4 to op. in crowded areas, up to 2 to 3 mm. in length and

THE ASPERGILLUS FLAVIPES GROUP 181

8 to 10/x in diameter in some strains and under some conditions of culture,
walls more or less yellow under the microscope, with color mostly localized in
the outer layers of the cell-wall and occasionally as disk-like concretions
on the surface of walls otherwise smooth. Vesicles subglobose to elliptical
(fig. 51 D), up to 30 by 4G> in the largest forms, usually with diameter
twice that of the conidiophore in smaller forms. Sterigmata in two series,
colorless or nearly so, closely packed over the apex of the vesicle in small
heads, and covering the vesicle in large heads, primary sterigmata about
6 or 8m by 2 to 3/x, secondary sterigmata 5 to 8/x by 1.5 to 2/x. Conidia 2
to 3ju, smooth, subglobose, colorless or nearly so under high magnification,
with chains aggregated to form columns as seen with the handlens in old
cultures.

Cosmopolitan in distribution and particularly common in soil and upon
decomposing organic materials.

Historically, the name applied to the series is taken from strains 4640.474
and 4640.402 (Thorn Collection) obtained through daFonseca from the
Bainier collection in Paris in 1922. These strains showed smooth yellow
con idiophores 300 to 400m by 3 to 4ju, contrasting with heads that were
rath er persistently white and possessed the general morphology of the series
as described above. Hulle cells were not found. Nevertheless, the close
relationship of these organisms to a great series of cultures obtained from
many sources in which these structures are regularly found justifies us in
broadening the use of the name.

Culture No. 4640.486 (Thorn) received from the Bainier collection as
S. rubescens (XRRL No. 291) shows deep floccose colonies with few heads
and scattered dark Ivyphal masses in age. Hulle cells have not been seen in
this strain.

Among forms commonly obtained from soils collected from widely
scattered areas in this country and abroad, a series of isolates seems to
comply with the description given by Blochwitz for Aspergillus archi-
flavipes (Ann. Mycol. 32(1/2): 84. 1934). This species as described
represents an extreme development toward radiate heads, abundant conidia,
conidiophores 2 to 3 mm. in length and the development of deep brown or
actual red shades of color in the mycelium. Several strains observed in
culture approach this description (fig. 51 C). Heads are at first globose,
then become slowly barrel-form, i.e., short stocky columns. Large brown
drops of transpired fluid are commonly seen which become yellow when
acidified and return to reddish shades when alkali is added. Recognition
of the species does not appear warranted since no clearly definable character
exists which distinguishes these isolates from the less colored forms generally
considered as representing Aspergillus flavipes in a more restricted sense.

182 A MANUAL OF THE ASPERGILLI

Antibiosis

Working with a culture of Aspergillus flavipes from Thorn (No. 4303.46),
NRRL No. 287, White (1943) has recently demonstrated the production of
an antibacterial substance which in its action against Staphylococcus aureus
strongly resembles penicillin. The substance was produced in greatest
amount in a medium containing 5 to 10 percent corn steeping liquor as the
sole nutrient. The addition of sugar is reported as definitely deleterious.

Occurrence and Economic Importance

Members of the Aspergillus flavipes group like hose of the A. ustus,
versicolor, and terreus groups are cosmopolitan in distribution, and are
especially common in fertile soil and upon decaying vegetation. They are
not known to be active agents of decomposition but are capable of growing
in the presence of a limited amount of water, hence are probably significant
in initiating or continuing processes of decay where most micro-organisms
are incapable of growing. Little is known of the biochemical activities or
products of these forms.

Chapter XIV
THE ASPERGILLUS VERSICOLOR GROUP

Outstanding Characters

Conidial heads hemispherical to almost globose, in many different shades

but usually showing green or blue-green.
Conidiophores smooth, colorless, more or less sinuous.
Vesicles globose to ovate or elliptical with radiate sterigmata borne over

the upper half to three-fourths of the surface.
Sterigmata in two series.
Spores globose or subglobose, echinulate.
HiiHe cells found in occasional strains, globose.

Members of the Aspergillus versicolor group are cosmopolitan in distribu-
tion. They occur regularly in soil, upon decaying vegetation, upon stored
grains, upon cured meats, and upon a multitude of other products exposed
to occasional moist air or undergoing slow decomposition. The morphology
of all strains (with the exception of Aspergillus j anus) is basically alike, but
different strains vary tremendously in their cultural appearance. This is
especially true of Aspergillus versicolor where conidial heads in different
strains vary in color from dark blue-green through green to yellow-green,
to yellow and orange, and finally in some strains to yellowish-cream, buff or
flesh color.

Group Key

I. Heads typically globose, less commonly hemispherical; blue-green in color with
the blue element dominant; colony reverse and substratum usually in red or
maroon shades A . sydowi series

A. Conidial heads always blue-green, globose to radiate

A. sydowi (Bain, and Sart.) Thorn and Church

B. Conidial heads of two types: (a) blue-green in color, vesicles subglobose to

elongate, borne on short conidiophores, and, (b) white, clavate, borne
upon long conidiophores A. janus Raper and Thom

II. Heads hemispherical or nearly globose at times; in green shades without blue
admixture, buff to orange-yellow or occasionally flesh colored; colony reverse
usually in pink, yellow-red or purple-red shades, rarely almost colorless

A. versicolor series

A. Conidia echinulate A. versicolor (Vuill.) Tiraboschi

B. Conidia smooth A. humicola Chad, and Sach.

183

184

A MANUAL OF THE ASPERGILLI

Aspergillus sydowi (Bain, and Sart.) Thom and Church, The Aspergilli,

p. 147. 1926.

Synonym: S. sydowi Bainier and Sartory, in Ann. Mycol. 11: 25-29, PI.
Ill, 1913. Compare PI. IV, cultures No. 4235.17 and K22,
Thom and Church, The Aspergilli. 192(5.

Colonies upon Czapek's solution agar growing well at room temperature,
in most strains close-textured and velvety from crowded conidiophores and
heads arising from the substratum (fig. 53 A), in other strains more or less

■ *\

■

-

1

i # « *

o

1 ~"' r*

•■* H

\
A

Fig. 52. Conidial structures in Aspergillus sydowi, NRRL No. 250, X 800. A,
Typical, large conidial head borne upon an erect conidiophore arising from the
substratum. B, Small head, borne as a lateral branch on an aerial hypha, consisting
of a small cluster of double sterigmata. C, Minute heads in which sterigmata appear
in a single series only.

floccose (fig. 53 B) from interlacing and trailing aerial hyphae bearing con-
idial heads ranging from large, well-formed structures to minute fruits
consisting of clusters of simple sterigmata bearing few conidial chains, blue-
green in color, approximately Delft blue or deep Delft blue (Ridgway, PI.
XLII) with the blue effect especially marked in young fruiting areas (PL
V A), reverse usually in shades of red, from coral red to maroon (Ridgway,

Plate V

A^feS »S?&*,aSaCB&S SK^iS0"^ C,hUr?h^R.RL N°- 25°- * C«PP- right)
1'iraboschi, NRRL No 239 D fcen e'r riJhn j.„ u ^ fcente-i- 1-ef t ) , AapnyiUiu versicolor (Vuill
Clower left), Aspergillus te w,fi NRRL v?^ "1t»''S"'° °'' tVll,,1U T/rab^chi, NRRL No. 233. £
Tins objective could not be attainedTn making theit iS "^ ""gn^uW be in cinnamon shades,
(v. Tiegh.) Blochwitz, VrSSO^^^^^^^T^ aJPT* ^^ f**"**" """""
were encountered in reproducing the colore i 7th stZ ) K.,™ S uS vinaceous fawn. Difficulties
all other cultures growing upon CWk's sol ht inn »i« r> i f ?' cult"re ««>wing on malt extract agar;
Research Laboratory. Kduc£P^ Northern Regional

THE ASPERGILLUS VERSICOLOR GROUP

185

Pis. XIII and I) to almost black in some strains in age. Conidial heads
typically radiate to nearly globose (fig. 52 A), ranging from 100 to 150m,
but often reduced to small penicillate clusters of sterigmata especially in
marginal colony areas and upon aerial hyphae (fig. 52 B and C). Conidio-

Fig. 53. Colony types in the Aspergillus versicolor group, on Czapek's solution
agar, 10 days, room temperature. A, Typical heavy sporing strain of A. sydowi.
B, Floccose, lighter sporing strain of the same species. C,A. versicolor, XRRLXo.
239, characterized by abundant green conidial heads and the production of deep
red exudate. D, A. versicolor, XRRL Xo. 227, characterized by flesh-colored conidial
heads.

phores mostly arising from submerged hyphae, up to 500m in length by 5
to 8m in diameter, colorless, smooth, comparatively thick-walled. Vesicles
nearly globose, fertile over almost the entire surface, up to 20m in diameter.
Sterigmata in two series; primary 6 to l\x by 2 to 3m, secondary 7 to 10m by
2 to 2.5m- Conidia globose 2.5 to 3.0m (Bainier), in our culture up to 3.5m
in diameter, conspicuously spinulose, green en masse. Globose hulle cells

186 A MANUAL OF THE ASPERGILLI

closely resembling those of the A. nidulans group have been seen in occa-
sional strains. No sclerotia or perithecia are found, but clamydospores in
solid substrata are reported.

A typical member of our series of isolations was sent to Bainier who con-
curred in our interpretation of his species.

Reduced conidial apparatus appears in varying degree in all strains of
A. sydowi examined. In typical strains, primary sterigmata, with their
clusters of secondaries each bearing a chain of conidia, are found singly
or variously grouped along trailing aerial hyphae. In other strains there is
a progressive development of aerial mycelium, in the form of trailing hyphae
either single or in ropes, coupled with a reduction in the number of typical
A. sydowi heads. Strains are even occasionally seen in which only a few
A. sydowi conidiophores and heads are found in what is otherwise a penicil-
lium-like colony. Thus a series of strains exists which shows a fairly com-
plete gradation from Bainier and Sartory's Sterigmatocystis sydowi to
Penicillium restrictum of Gilman and Abbott. We are led to believe that
the latter species should probably be assigned to this section of the genus
Aspergillus.

While morphologically very close to A. versicolor, A. sydowi is easily pre-
sumptively recognized by the characteristic blue-green color of its conidial
heads and the red colors in the substratum. A partial list of the sources of
the isolations studied includes soil in Washington, D. C, Illinois, Manitoba,
Florida, and Ceylon; moldy silk from a stocking factory; concentrated sugar
products from Louisiana; dried fish in Japan; and bee-hives in Michigan.
It is world-wide in distribution and very adaptable to substrata of widely
different nature.

Probable Synonyms

Aspergillus tiraboschii Carbone (Atti d. Inst. Bol. Univ. Pavia Ser. II. Vol. XIV,
p. 320, 1914) is described in the colors of A. versicolor but with the head of A. sydowi.
It would appear to be more or less intermediate but, unless reisolated, must be
dropped because of incomplete data.

Sterigmatocystis tunetana Langeron (Bull. Soc. Path. Exot. 17: 345-347, text fig.,
1924) . This mold was recorded as isolated from an ulcer of the hand but failed to
produce lesions in animal tests; as described the colonies were blue-green as in
A. sydowi.

A. sydowi var. achlamydosporus Nakazawa, Simo and Watanabe (Jour. Agr. Chem.
Soc. Japan 10(2): 178-179. 1934). The absence of chlamydospores (htille cells ?)
in a strain of A. sydowi is hardly a sound basis for separation.

Sterigmatocystis cyaneus Mattlet (Ann. Soc. Belg. Med. Trop. 6: 32, 1926) was
described without data to separate it from A. sydowi.

S. cameleo Sartory, Sartory, and Meyer (Ann. Mycol. 29: 360-361, PI. Ill, figs. 7-8.
1930) by description must have been some strain of A . sydowi although the very small
smooth conidia do not agree. It may possibly represent a form near A. humicola as
described by Chaudhuri and Sachar (see p. 193).

THE ASPERGILLUS VERSICOLOR GROUP 187

Aspergillus jamts Raper and Thorn, in Mycologia 36 : 55G-561, fig. 1. 1944.

Species characterized by conidial heads of two distinct types, (1) large
white heads borne upon long conidiophores terminating in strongly clavate
vesicles and (2) smaller, dark green heads borne upon short conidiophores
with typically ovate vesicles (PI. I E and F, and V B).

Colonies varying greatty in color and in texture depending upon the sub-
stratum and the temperature of incubation. Upon Czapek's solution agar
at 24° C. (fig. 54 A) colonies spreading irregularly, usually consisting of a
central floccose mass 1 to 2 mm. deep, pale yellow-buff in color, bearing few
and scattered fruiting structures surrounded by an irregular zone of
crowded fructifications with dark green heads occurring in a dense stand
adjacent to the substratum (fig. 54 D) and with numerous long-stalked
white heads projecting above this layer (fig. 54 C); reverse in dull yellow to
light brown shades. When incubated at 20° C, colonies more restricted,
less floccose and consisting almost exclusively of a dense stand of long-
stalked white heads with small green heads absent or developing only in age,
and arising from trailing aerial hyphae entwined among the white fruiting
structures. When incubated at 30 to 32° C. colonies close-textured, pre-
dominantly green but with central area commonly showing irregular patches
of massed hiille cells, buff to dull yellow in color. Conidial heads abundant
and consistently dark green in color. Reverse in dull brown shades.

White conidial heads loose in texture (fig. 54 C), consisting of radiating
and divergent chains of conidia, eommonfy 150 to 200m in diameter, occa-
sionally larger. Conidiophores long, thin, mostly 2 to 2.5mm. in length by
8 to 10.5m in diameter, occasionally larger, erect, essentially uniform in
diameter throughout but often marked by numerous and irregularly spaced
constrictions, walls smooth, colorless, approximately 1 to 1.4/z in thickness.
Vesicles thin-walled, clavate (fig. 54 G), mostly 45 to 60m by 15 to 18m with
individual structures larger or smaller, entire surface loosely covered by
sterigmata as a rule, but often showing barren areas which may occupy any
part of the sterigmatic surface. Sterigmata in two series, primaries 7 to
10m by 3.5 to 4.5m; secondaries 6 to 8m by 2.5 to 3m. Conidia smooth, color-
less, globose to subglobose, mostly 2 to 2.5m, with maximum about 2.8m.

Green conidial heads compact, radiate when young, becoming columnar
in age and often spreading into two divergent columns. Heads at first in
blue to blue-green shades near dark gobelin blue (Ridgwav, PI. XXIV),
becoming dark olive-gray in age (Ridgway, PI. LI), in size commonly rang-
ing from 60 to 75m in diameter to 200 to 300m in length. Conidiophores
erect, commonly 300 to 400m in length, by 6.5 to 8m in diameter, of uniform
thickness throughout, walls smooth, colorless or very faintly green, approx-
imately 1 to 2m thick, enlarging rather abruptly into an ovate vesicle. Ves-
icle thin-walled, variable in form and dimensions, but commonly ovoid (fig.

i o

l\

i

Fig. 54. Aspergillus janus; NRRL No. 1787. A and B, Colonies on Czapek's
solution agar and malt agar, respectively, showing a, scattered white conidial heads,
b, crowded green heads, and c, massed niille cells, two weeks old, incubation at 2-1°

C, X i. C, Characteristic white heads borne on long thin conidiophores, X 7.5.

D, Massed green heads in dense stand adjacent to the substratum, with white heads
projecting from the left, and with localized development of hulle cells in two limited
areas at right, X 7.5. E, Heads of mixed character with older (outer) conidia white
and younger (inner) conidia green, X 24. F, Hulle cells, X 350. G, Typical white
head showing clavate vesicle, double sterigmata and small, smooth conidia, X 450.
H, Head of mixed character, at this stage (early) producing small white, smooth
conidia, X 450. /, Typical green head, showing small, nearly globose vesicle, double
sterigmata and larger, echinulate, green conidia, X 450. (Reprinted from Raper
and Thorn, "New Species of Aspergilli from Soil," Mycologia 36: Nov -Dec. 1944.)

188

THE ASPERGILLUS VERSICOLOR GROUP 189

54 I) and occasionally conspicuously elongate, typically fertile over the
entire area, ranging in size from 20 to 30/x by 12 to 18/z. Sterigmata in
two series, rather loosely arranged, primaries 7 to 10m by 4 to 4.5m; sec-
ondaries 6 to 8.5^ by 2 to 2.8m- Conidia dark green in mass, conspicuously
spinulose (fig. 54 I), globose, mostly 2.5 to 3.5/x, occasionally larger or
smaller.

Conidial heads of mixed character, containing both white and green
spores, commonly encountered (fig. 54 E), usually borne upon long conidio-
phores approaching and often equalling in length those of white heads,
vesicles clavate (fig. 54 H), sterigmata at first bearing colorless smooth-
walled conidia, but subsequently bearing dark green spinulose conidia. At
temperatures of 24° C. and above, thick-walled hulle cells abundant, irregu-
lar in form (fig. 54 F), commonly globose to subglobose, not infrequently
elongate, commonly more or less curved and often lobed.

Colonies upon malt extract agar growing luxuriantly (fig. 54 B), generally
loose in texture with aerial mycelium prominent, conidial heads normally
more abundant than upon Czapek agar, the proportion of white to green
heads varying with the temperature of incubation.

Colonies upon hay infusion agar spreading broadly, consisting of a thin
submerged mycelium from which develop erect, white and green conidial
structures, the relative proportion of these types being dependent upon the
temperature of incubation ; since comparatively meager growth occurs upon
this medium, and since there is a minimum of aerial vegetative hyphae, it
constitutes a very favorable substratum upon which to observe the forma-
tion of the contrasting fruiting structures characteristic of the species.

The binomial Aspergillus janus was selected for this species because of the
contrasting types of conidial heads produced — it is literally a "two-faced"
mold.

Type culture NRRL No. 1787 was isolated in February 1942 from
Panama soil collected during the summer of 1941 by John T. Bonner of
Harvard University. Three additional isolations by members of the
Northern Regional Research Laboratory staff have since been made from
Panama soils subsequently collected by Mr. Benjamin T. Coghill.

It is believed that this species represents a normal component of the
microflora of Panama. Additional evidence in support of this view is
furnished by the fact that in 1925 Professor Roland Thaxter sent to Thorn
under the label "white Panama Aspergillus" a representative of this species.
The form was never described by Thaxter, and viable cultures of it were lost
from our collection some time prior to 1930. As the correspondence of the
time is remembered, Thaxter was plagued by the presence of a small green
mold which repeatedly appeared in his cultures as a "contaminant."
Fortunately, the original tube received from Thaxter has been preserved,

190 A MANUAL OF THE ASPERGILLI

and re-examination of this culture leaves no question but that he was deal-
ing with a strain of the species here described, and that the green form
which troubled him so much was not, in fact, a contaminant but a different
phase of the same fungus.

Aspergillus janus var. brevis Raper and Thorn, in Mycologia 36 : 561-563,

fig. 2. 1944.

The variety differs from the species in a number of particulars, foremost
among which are (1) the reduced length of the conidiophores bearing both
white and green heads and (2) a consistent tendency for white and green
conidial structures to develop in approximately pure stands and to appear
as contrasting radial sectors.

White conidial heads are of the same general pattern and form as in the
species, but are of somewhat smaller dimensions, are borne upon conidio-
phores generally less than 2 mm. in length by 6 to 8m in diameter, and are
characterized by elongate but not strongly clavate vesicles measuring 20
to 25^ by 14 to 18m; conidia are smooth-walled, colorless, globose to sub-
globose, 2.2 to 2.8m in diameter. Green conidial heads are compact, glo-
bose to somewhat columnar, borne upon conidiophores 75 to 125m by 4 to
6m with globose to subglobose vesicles measuring 8 to 15m by 10 to 18m;
conidia are dark blue-green, strongly echinulate, and 3.5 to 4.5m in diameter.

The vesicles of white heads in the variety brevis are of approximately the
same size and form as the vesicles of green heads in the species itself,
whereas the conidiophores bearing each type of head are approximately
one-half the length of those bearing the same type of head in the species.
The most striking character distinguishing the variety, however, is the
manner in which areas of white and green heads are sharply separated along
radial lines. Conidial heads of mixed character are produced and usually
can be found along the frontier between white and green sectors.

Type culture NRRL No. 1935 was isolated in July 1942 from a sample of
soil collected in Alameda in southern Mexico, and forwarded to us in June
by Mr. William B. Roos.

Aspergillus versicolor (Vuill.) Tiraboschi, in Ann. Botan. (Rome)

7:9. 1908.

Synonym: S. versicolor Vuillemin. Mirsky, B., in These de Med.
Nancy, no. 27, p. 15 et seq. 1903. See Thorn and Church,
The Aspergilli, p. 142, 1926.

Colonies upon Czapek's solution agar rather slow growing, compact, in
some strains velvety and consisting almost entirely of closely crowded
conidiophores arising from the substratum, in other strains showing a
marked development of floccose hyphae bearing more or less abundant

THE ASPERGILLUS VERSICOLOR GROUP

191

conidiophores as short aerial branches, in still others a combination of both
growth types with colony centers initially floccose and outer areas almost
velvety, at first white, passing through shades of yellow, orange-yellow, tan,

Fig. 55. Aspergillus versicolor, strain NRRL Xo. 227. Photomicrograph showing
details of head structure. Note particularly the double series of sterigmata, X 2100.

to yellowish-green shades such as pea green or sage green (Ridgway, PI.
XLVII) depending upon the strain (PI. V C and D, and fig. 53 C and D) and
conditions of culture, occasionally with the green colors almost or com-

192 A MANUAL OF THE ASPERGILLI

pletely lacking; reverse and substratum occasionally colorless or nearly so,
mostly passing through shades of yellow to orange then rose, purple-red or
red, the particular shade and intensity of color normally persisting as a
strain or varietal character. Heads roughly hemispherical, radiate, up to
100 to 125m in diameter; rarely approaching columnar. Conidiophores
colorless, smooth, up to 500 or even 700m by 5m or approaching 10/x near the
vesicles. Vesicles 12 to 20m in diameter, fertile area hemispherical or semi-
elliptical (fig. 55) passing almost imperceptibly into the funnel-like enlarged
apex of the conidiophore. Sterigmata in two series, primary commonly 8
to 10/x by 3m, occasionally less, secondary 5 to 10m by 2 to 2.5m. Conidia
globose, usually delicately echinulate, mostly 2.5 to 3m, occasionally 3.5m or
4m, usually borne in loosely radiating chains.

In occasional highly colored strains vesicles and sterigmata may be
colored.

Hiille cells of the Aspergillus nidulans type are occasionally seen.

Neither perithecia nor sclerotia have been found.

The diagnosis is drawn broadly enough to cover a common and very
abundant series of organisms which vary greatly in colony appearance. In
some strains aerial growth is composed of conidiophores and heads only; in
others it is made up of floccose felts or ropes of hyphae bearing short conidio-
phores and small heads which retain the characteristic arrangement of
vesicles, sterigmata, and conidial chains.

The wide range of colony structure and coloration of strains,1 and the
varied conditions under which these have been collected probably account
for the appearance of many names in the literature which refer to cultures of
the group but which cannot be safely separated and identified by the de-
scriptions given.

The following list of species are believed to represent synonyms:

S. ambari Beauregard (Ann. de Micrographie 10: 255-278, pi. 1. 1898). Probably
one of the series, but never identified again.

S. bicolor J. Ray (Rev. Gen. Bot. 9: 193-212, 245-259, 282-304, PI. 12-17. 1897)
is probably one of this series. Primary sterigmata were reported filled with red
coloring matter and conidia globose, spinulose, up to 2.5ju in diameter.

Sterigmatocystis brodeni Mattlet (Name only in Ann. Soc. Beige Med. Trop. 6:
31, 1926; discussion, ibid 4: 167-171, figs. 1, 2. 1924). Apparently a member of
this series.

A. flavo-viridescens Hanzawa (Jour. Coll. Agr. Tohoku Imp. Univ. Sapporo 4:
232-3, pi. 21, figs. 1-4. 1911). Culture No. 4291.10 (Thorn) received from Hanzawa
under this name belongs to the A. versicolor group.

A. versicolor var. glauca Blochwitz (Ann. Mycol. 32(1/2): 86. 1934). This variety
has the color of the A. glaucus group which is more deeply blue than A. versicolor.
Isolated in the skin clinic at Kiel upon human skin showing "ringworm.'

>>

1 For a more complete discussion of the range of cultural types found in this series,
the reader is referred to Thorn and Church, The Aspergilli, pp. 142-145, 1926.

THE ASPERGILLUS VERSICOLOR GROUP 193

8. glauca Bainier (Bui. Soc. Bot. France 27: 29-30, pi. 1, fig. 3, 1880, ibid. 28: 77,
1881). From extract of henbane, dregs of wine, casks, and corks.

A. globosus Jensen (N. Y. Cornell Agr. Exp. Sta. Bui. 315, p. 482, 1912). The
type culture received from Whetzel (Thom No. 2705) is certainly a member of this
series, and is characterized by yellowish-green to olive-green conidial areas, with
colony reverse in yellowish-orange to wine red.

S. polychroma Ferraris (Fl. Ital. Crypt. Hyph. p. 640). Syn. A. versicolor, fide
Tiraboschi, in Ann. de Botanica (Rome) 7: 9, 1908.

S. spuria Schroeter (Cohn Krypto. Fl. von Schlesien 3: 2 Halfte, Lief. 1, p. 218,
1893). Position in doubt. May represent a form of A. versicolor similar to the flesh
colored forms discussed by Thom and Church in The Aspergilli, p. 145, 1926, or may
belong with A. carneus (See p. 201).

A . tabacinus Nakazawa, Simo, and Watanabe (Jour. Agr. Chem. Soc. Japan 10(2):
177-178, 1934). The detailed figures given in contrast to their own strain of A. versi-
color showed differences which disappear when large numbers of isolations are
studied.

Aspergillus humicola Chaudhuri and Sachar, in Ann. Mycol. 32: 97. 1934.

Characterization after Chaudhuri and Sachar

Colonies on Czapek's solution agar, at first white passing through shades
of olive-gray (Ridgway, PI. XLVI. 20.o_yy) velvety at margin, floccose
toward the center; reverse and substratum in shades of yellow, heads radi-
ate. Conidiophores arising directly from the substratum, up to 300m in
length by 4 to 5.4m in diameter, or as short branches, about 70m long, from
aerial hyphae; walls smooth and almost colorless. Vesicles 9 to 15m in
diameter, colorless, flask-shaped, with sterigmata radiating from the whole
surface of the larger heads, or only borne in the upper third in small heads;
primary sterigmata 3.6 to 5.4m by 1.8 to 2m; secondary sterigmata 3.6 by
1.8m- Conidia globose, smooth, 2 to 3m in diameter, in radiating chains.

Neill (1939) is believed to have correctly placed this organism with A.
versicolor and its allies despite the smoothness of its conidia. The "almost"
colorless conidiophore suggests relationship to Aspergillus ustus; in this
group certain forms (e.g., Blochwitz's A. ustus var. laevis) apparently have
spores smooth or nearly so (see p. 175).

Pathogenesis

Aspergillus sydowi is not reported by name as a parasite, but strains
described in terms which must be interpreted as placing them with A. sydowi
includes A. tunetanus (Langeron) Dodge from fleshy lesions on a hand in
Tunis: A. Vancampenhouti (Mattlet) Dodge also from tropical Africa; A.
cyaneus (Mattlet) Dodge from the same region. The evidence at hand
links A. sydowi more closely with A. nidulans than was formerly supposed,
although such relationship is strongly indicated by the identical character
of their hulle cells. The fragmentary descriptions commonly given for

194 A MANUAL OF THE ASPERGILLI

individual pathogenic molds isolated by persons unfamiliar with the litera-
ture are rarely definite enough to separate nearly related forms.

An occasional culture of A. versicolor is obtained from apparently patho-
genic sources. Thus far experimental work has not shown evidence of
actual lesions in human flesh. Little colonies bearing green heads and
conidia were drawn with a breast pump from an inflamed mammary gland
and kept in culture for many years but failed to grow in laboratory media at
blood heat, Aspergillus versicolor var. glauca Blochwitz was isolated from
human skin showing "ringworm" at the skin clinic in Kiel, but apparently
pathogenicity was not proved experimentally. Strains of A. versicolor are
frequently observed upon dried salted lean beef, thus showing its capacity
to grow in and upon meat products, but not giving direct evidence of par-
ticipation in any pathological process.

Occurrence and Economic Importance

Members of the Aspergillus versicolor group appear widely distributed in
soil, on spoiling and drying food stuffs, breads, cereals, old cheese, dried
meats, cured India rubber, musty vegetable products, and other substrata
characterized by a moderately low water content or containing factors toxic
to most organisns. They are reported as capable of decomposing certain
paraffins. The production of proteolytic enzymes by most strains is shown
by the digestion of milk and the liquefaction of gelatin. Aspergillus
sydowi, in particular, is a characteristic component of all soil examined.

Fat production by .4. sydowi has been studied quite extensively by Pro-
fessor Peterson and associates at the University of Wisconsin. For reference
to this work see the various papers listed in the Topical Bibliography under
the heading "Chemistry of Mold Tissues."

Chapter XV
THE ASPERGILLUS TERREUS GROUP

Outstanding Characters

Heads columnar, in cinnamon, pale buff or light flesh colors.
Conidiophores smooth, colorless, rarely exceeding 250m in length.
Vesicles hemispherical, with upper half to two-thirds covered by sterig-

mata.
Sterigmata in two series, generally crowded.
Conidia smooth, globose to slightly elliptical, small.

Included here are members of a variable and cosmopolitan group of
Aspergilli especially common in soil. They differ markedly in color and in
colony appearance and to a lesser degree in the texture of their conidial
heads. The group may be separated as follows:

Group Key

I. Conidial heads in cinnamon or orange-brown shades, compact, uniform in di-
ameter throughout A . lerreus series

A. Colonies velvety, conidiophores mostly in a dense stand arising from the

substratum.

1 . Conidial heads in dull cinnamon shades A . lerreus Thorn

2. Conidial heads orange-brown near xanthine orange (Ridgway)

A. lerreus var. boedijni (Bloch.) n. var.

B. Colonies floccose, conidial heads arising from aerial hyphae.

1. Mycelium colorless, heads light pinkish-cinnamon in color

A . lerreus var. floccosus Shih

2. Mycelium yellow, heads developing late, in cream or light tan shades

.4. terreus var. aureus n. var.

II. Conidial heads white or flesh colored, loose textured not strictly uniform in
diameter A . carneus series

A. Colonies in light flesh colors, ranging from near white to vinaceous-fawn

(Ridgway). Thick-walled hyphae suggestive of hiille cells are generally
present A. carneus (van Tieghem) Blochwitz

B. Colonies persistently white or becoming dull ivory in age. Thick-walled

cells often present A. niveus Bloch.

Aspergillus terreus Thorn, in Turesson, Gote, Svensk Botanisk Tidskrift

10: 5, 1916, without description; diagnosis Thorn and Church,

in Amer. Jour. Bot. 5: 85-6. 1918.

Synonym: A. galeritus Blochwitz, in Ann. Mycol. 27(3/4): 205. Taf.
III. 1929.

Colonies upon Czapek's solution agar growing well at room temperature
and up to 37° C, spreading, plane or marked by shallow radial furrows,

195

196

A MANUAL OF THE ASPERGILLI

Fig. 56. Aspergillus terreus. A, Colony margin of a typical strain, NRRL No.
265, showing crowded, long columnar heads, X 14. B, Scattered but typical conidial
heads of an ultra-violet light-produced mutation unable to utilize NO3 nitrogen,
X 50. Both cultures on Czapek's solution agar, 2 weeks.

THE ASPERGILLUS TERREUS GROUP 197

velvety (fig. 58 A) or in some strains showing tendency toward floccosity in
central colony areas, heavy sporing throughout with massed columnar heads
giving to colonies their characteristic color and texture, in color ranging
through various cinnamon shades (Ridgway, PI. XXIX) depending upon
the abundance and maturity of the heads (PI. V E), amber exudate produced
in some strains, odor transient to none, reverse in dull yellow to brown
shades. Conidial heads long columnar (fig. 56 A), with conidial chains
compacted together, of uniform diameter throughout entire length, com-
monly ranging from 150 to 500m or more in length by 30 to 50m at maturity,
ranging from cinnamon-buff through cinnamon to Sayal brown (Ridgway,
PI. XXIX). Conidiophores more or less flexuous, smooth, colorless, com-
monly ranging from 100 to 250m by 4.5 to 6.0m, approximately uniform in
width throughout (fig. 57 A and 19 A). Vesicles hemispherical, dome-like,
commonly 10 to 16m in diameter, merging almost imperceptibly into the
supporting conidiophore. Sterigmata in two series, primaries crowded
(fig. 57 A), parallel, 5.0 to 7.0m by 2.0 to 2.5m, secondaries closely packed
5.5 to 7.5m by 1.5 to 2.0m- Conidia globose to slightly elliptical, commonly
1.8 to 2.4m in diameter.

Species description based upon type strain NRRL No. 255 (Thorn No.
144) and innumerable additional isolations from soils and other sources in
this country and abroad. The species is especially abundant in warm and
comparatively dry arable soils.

The great majority of isolates belonging to this series fall within A.
terreus in its strictest sense, and duplicate in all essential particulars the
description given above for this species. Nevertheless, wide natural varia-
tion among strains is encountered when great numbers are isolated from
widely separated sources. Some of these are quite striking in appearance
and have apparently furnished the bases for species and varietal description
by other workers.

Aspergillus terreus var. boedijni (Bloch.) n. var.

Blcchwitz, in Ann. Mycol. 32(1/2): 83, 1934, described Aspergillus
boedijni as a new species differing from Aspergillus terreus primarily in the
color of its conidia. These were reported as pure yellow at first, becoming
pure brown or ochraceous-brown in age, and brighter than .4. galeritus
Blochwitz.1 In our experience strains are occasionally encountered which
are characterized by a bright, orange-brown color instead of the dull cinna-

1 A. galeritus Blochwitz (Ann. Mycol. 27(3/4) : 205, Taf. Ill, 1929) is a redescription
of A. terreus Thorn. Blochwitz acknowledged having Thorn's type at hand when
he renamed this species. No reason was given beyond the claim that he had had the
organism in culture for some years before Thorn's description oi' A. terreus was
published.

198 A MANUAL OF THE ASPERGILLI

mon shades typical of the species. Among cultures currently under exami-
nation, this character is noted in an isolate from Argentine soil, is somewhat
more marked in NRRL No. 680 from Dr. G. A. Ledingham, Ottawa,
Canada, and is particularly striking in NRRL No. 1913 isolated by Dr. C
W. Emmons from Arizona soil. In all of these strains the basic morphology
is that of a typical A. terreus, hence Blochwitz's separation is reduced to
varietal rank.

Aspergillus terreus var. floccosus Shih, in Lingnan Sci. Jour. 15 : 372, PI. 16,

fig. 3, 1936.

Strains characterized by deep floccose colonies in which conidial heads
are less abundant, develop late, and are borne almost entirely upon aerial
hyphae, are frequently encountered (fig. 58 B). While the conidial struc-
tures of certain of these strains appear entirely normal, in the majority of
isolates the heads are somewhat less compact and generally lighter in color.
This color difference is particularly marked among isolates from soils col-
lected in Texas, Central America, and Cuba with color commonly light
pinkish-cinnamon (Ridgway, PI. XXIX) to vinaceous-buff (PL XL) in age.
No sharp line of separation can be drawn between typical strains of A.
terreus and the floccose forms under consideration since isolates of interme-
diate character are encountered; nevertheless, these strongly floccose
cultures occur with sufficient frequency to warrant recognition of Shih's
varietal designation if his interpretation is somewhat broadened. The
variety is considered by the writers as a strongly floccose Aspergillus terreus
in which the head is commonly less compact and lighter in color, but with
the basic morphology of the conidial apparatus remaining that of the species
proper. This variety is represented by such strains as NRRL Nos. 1920
and 1921, isolated from Cuban soil contributed by Professor J. M. Osorio,
University of Havana; No. 1922, isolated from Texas soil, collected and sent
to us by Dr. F. E. Clark from Greenville, Texas.

Other strains examined almost completely bridge the gap between the
pale colored strains of A. terreus var. floccosus and the light flesh colored
forms characteristic of Aspergillus carneus.

Aspergillus terreus var. aureus n. var.

This new and striking variety differs from the species in a number of par-
ticulars: colonies upon Czapek's solution and malt extract agars are com-
paratively slow growing, floccose, ranging up to 3 to 4 mm. deep, and are
bright golden yellow in color. Conidial structures are produced tardily and
in limited numbers. Conidiophores are appreciably longer than those of
the species, often becoming 500m or more in length, and bear columnar
heads, generally loose in texture, ranging in color from cream or light buff

THE ASPERGILLUS TERREUS GROUP

199

to light pinkish-cinnamon. Microscopically, the conidial structures ap-
proximate those of the species itself. Separation as a new variety is based
primarily upon the characteristic coloration of the growing colony.

Fig. 57. Conidial structures of members of the Aspergillus terreus group, X 840.
A, A. terreus, type strain NRRL No. 255 (Thorn No. 164). B, A. terreus var. aureus,
NRRL No. 1923. d, C2, and C3, A. carneus, NRRL No. 1928, conidial heads vary
greatly in size. Du D2, and D3, A. niveus, NRRL No. 515, conidial heads of varying
dimensions.

Type strain NRRL No. 1923 (fig. 16 D) was isolated from Texas soil
contributed by Dr. F. E. Clark. Additional strains showing approximately
the same cultural and morphological characteristics have been isolated
from soils collected in Arkansas and Arizona. In A. terreus var. aureus the
yellow coloring matter is lodged in the vegetative mycelium, and there are
no suggestions of hulle cells; in A. carneus, however, approximately the same

200

A MANUAL OF THE ASPERGILLI

yellow tints are developed through the massing in localized colony areas of
thick-walled hyphae suggestive of hiille cells (see p. 201).

At least two other species have been described which are believed to
represent probably synonyms of Aspergillus terreus:

Aspergillus fuscus Anions (Archief. voor de Suikerindustrie in Nederlandsch.
Indie Jaarg. 29, Deell, pp. 8-10, 1921) by description is obviously a form closely
related to, if not identical with, A. terreus.

Fig. 58. Aspergillus terreus group; different species growing upon Czapek s solu-
tion agar at room temperature. A, Typical, heavy sporing strain of A. terreus,
NRRL No. 265. B, A. terreus var. floccosus, characterized by loose floccose colonies
and limited spore production. C, A. carneus, NRRL No. 1928, heavy sporing and
characterized by flesh-colored conidia. D, A. niveus, NRRL No. 515, characterized
by white conidial heads.

Aspergillus cmnamominus (Weiss) Dodge, in Med. Myc, p. 627. 1935.

Synonym: S. cinnamominus Weiss, in Ann. Parasitol. Hum. Comp. 8: 189-193,
5 figs 1930.
Characterization from Dodge: Hyphae septate and branched; conidiophores
simple, 5m in diameter, vesicle 12 by 9M. Primary phialides cylindric, 4M long, bearing

THE ASPERGILLUS TERREUS GROUP 201

2 to 3, sometimes 4 secondary phialides, 5 to 6ju long. Conidia spherical, 1 to 2/x in
diameter, brown. Other spores, borne laterally on short branches (phialides ?),

3 to 4p. Chlamydospores occasional. Under some conditions, only monstrous
phialides are formed, with a "pleomorphic" mycelium resulting. Described as pres-
ent in lesions of pityriasis versicolor flava. Inoculation into the human skin repro-
duced the original disease. From the description given the organism is some strain
of the A. terreus group.

Aspergillus carneus (v. Tiegh.) Blochwitz

Synonyms: Sterigmatocystis carnea v. Tiegh., in Bull. Soc. Bot. France
24: 103. 1877. Cited also in Saccardo Sylloge 4: 74, and
in Wehmer's Monograph, p. 127 (Mem. Soc. Phys. Hist.
Nat. Gen. pp. 1-157. 1899-1901).
Sterigmatocystis spuria Schroter as a bibliographic change in
Colin Krypt. Fl. Schleisen 3 : 2 Helfte Lief. 1, p. 218. 1893.
Aspergillus carneus Blochwitz, in Ann. Mycol. 31(1/2): 81.
1933.

Colonies upon Czapek's solution agar growing well at room temperature
spreading, plane or radially furrowed, more or less floccose, at first white
but becoming pale vinaceous-fawn to vinaceous-fawn (Ridgway, PI. XL)
with the development of mature fruiting structures (PI. V F, and fig. 58 C),
ranging from 1 to 2 mm. or more deep in central areas to very thin and
spreading at colony margin, comparatively heavy sporing throughout with
fructifications arising from both aerial and submerged mycelium, some
strains showing limited areas yellow in color from an underlying felt of
heavy-walled sterile hyphae suggestive of hulle cells (fig. 49 F) ; odor often
pronounced, somewhat putrid; reverse in orange-yellow, bright orange, to
deep brown shades; conidial heads loosely columnar, averaging 150 to 200m
by 25 to 35m, but commonly somewhat larger, varying in color as the colony.
Conidiophores variable in length, mostly 250 to 400m but ranging up to 1
mm., occasionally bearing secondary fruiting structures as short and irregu-
larly placed branches, smooth, sinuous, uncolored, mostly 3.5 to 6.0m in
diameter. Vesicles hemispherical, ranging from 5.5 to 9m, rarely as much as
10m (fig. 57 Ci). Sterigmata in two series, primary 5.5 to 6m by 2 to 2.5m,
secondary 5 to 5.5m by 1.8 to 2m, commonly very few primary sterigmata
present. Conidia globose to subglobose, thin-walled, averaging 2.4 to 2.8m,
with maximum rarely exceeding 3.2m.

Colonies upon malt extract agar growing more restrictedly, heavier spor-
ing, with pigmentation generally more pronounced and with conidial heads
averaging slightly larger than on Czapek's solution agar, otherwise dupli-
cating the above description.

This species is represented by strains NRRL No. 527, isolated as an air
contaminant in Washington; NRRL No. 298, isolated from Kansas soil;
and other soil isolations from different parts of the United States, Mexico,

202 A MANUAL OF THE ASPERGILLI

Cuba, and Central America. Strains differing from the above in the
absence of any yellow, thick-walled hulle-like cells are occasionally isolated
from soils. NRRL No. 1928, isolated from Arkansas soil, is representative.
In the absence of any yellow component, the color of these strains is more
accurately described as pale to light grayish-vinaceous-fawn (Ridgway, PI.
XXXIX).

The name Aspergillus carneus is revived to cover the forms under con-
sideration since their most obvious identifying characteristic is the pale
flesh color of their massed conidial heads. It is the belief of the writers that
van Tieghem probably had in hand some member of this series when he
proposed the name S. carnea, although, due to the inadequacy of his descrip-
tion, it is now impossible to establish this point with certainty. In any case,
the name is excellently descriptive of strains commonly encountered, hence
its application in this connection.

In describing Aspergillus carneus as a new species, Blochwitz (1933)
acknowledged the earlier use of this specific name by van Tieghem but dis-
regarded its validity. He undoubtedly applied it to a member of the
species as it is considered by us since he noted that it differed from A.
terreus (A. galeritus) principally in the flesh to rose color of its conidia. It
is believed that Blochwitz's A. niveus var. nubila (1934) likewise represented
a strain of A. carneus characterized primarily by conidia of darker rose, a
condition which in older cultures is frequently suggested by NRRL No.
1928 cited above.

Oilman and Abbott in their "Summary of Soil Fungi" (1927) called atten-
tion to the repeated isolation, from Louisiana soils, of forms with the
"general morphology of the Aspergillus candidus group but producing bright
pink conidial heads." While the writers think the affinities of these forms
lie more with Aspergillus terreus (columnar heads, colorless conidiophores)
and Aspergillus fiavipes (elongate, irregular hiille cells) than A. candidus,
we have every reason to believe they were dealing with forms similar to
those here designated A. carneus.

S. albo-rosea Sartory, Sartory, and Meyer (Ann. Mycol. 28: 358-359, PI. Ill,
fig. 1-6, 1930) apparently represents a member of this series. This is indicated by
the described coloration of colonies and more particularly by the detailed measure-
ments cited for it.

Aspergillus niveus Blochwitz, in Ann. Mycol. 27(3/4): 205-6, fig. 2,

Taf. III. 1929.

Synonym: A. eburneus Biourge, name attached to a culture received by
Thorn (No. 5402.1 : NRRL No. 515).

Colonies upon Czapek's solution agar white, plane or radially furrowed,
rather slow growing, forming a dense felt of mycelium and conidiophores up

THE ASPERGILLUS TERREUS GROUP 203

to 600m to 1 mm. deep, thinning toward the margin (fig. 58 D) and com-
monly spreading in unevenly radiating lines, commonly producing abundant
amber to brown exudate ; reverse in dark yellow shades through greenish to
brownish-black; odor slight. Conidiophores smooth with Avails colorless,
sinuate, more or less septate, slender, 4 to 6m in diameter, enlarging to a
hemispherical vesicle 8 to 15m in diameter or sometimes larger at the apex,
commonly 300 to 600m in length, occasionally up to 1000m long, and on other
substrata sometimes longer. Conidial heads showing chains of conidia in
comparatively loose columns, most frequently 20 to 30m in diameter but in
large heads up to 60m, with the general appearance of a snow white A.
terreus. Vesicular area hemispherical (fig. 57 D). Sterigmata in two series,
primary sterigmata 5 to 8m by 2.5 to 3.0m, secondary sterigmata 5 to 7m by
2 to 2.5m- Conidia 2.0 to 2.5m, rarely more, smooth, thin-walled, colorless.

Represented in the NRRL collection by Nos. 515 (Thorn No. 5402.1),
288, and 1955. Repeatedly isolated from soil but less common than
Aspergillus carneus and the ubiquitous A. terreus.

Typically the conidial apparatus is that of a white, loosety columnar
form of A. terreus. As described by Blochwitz, it does not show yellow in
culture. This is true of strain XRRL No. 515, although in age this culture
reaches dull ivory to pale buff on agar slants. In other strains, such as
XRRL No. 1955 from Dr. Timonin in Ottawa, Canada, limited areas mav
become yellow from the development of massed thick-walled hyphae as in
A. carneus.

Sterigmatocystis pusilla Peyronel (I germi atmospherici dei funghi con micelio.
Thesis. Padova. 1931, p. 21) probably represents a synonym of A. niveus Blochwitz.
It was described as follows: Colonies white, very thin; sterile hyphae creeping,
sparingly branched, falsely septate, hyaline, 1.5 to 5/x in diameter; conidiophores
erect, unseptate, hyaline, 60 to 80m by 2.5 to 3m, with apical vesicles hyaline, obovoid
or subglobose, 7 to 10m in diameter, sterigmata radiate, in two series, with primaries
5 to 7/x by 2 to 2.5m and secondaries 3 to 5m by 2m in groups of 2 to 3; conidia globose
2 to 2.5m, hyaline, smooth. Habitat: from air in northern Italy at altitude of 1,700
meters. From description this would appear to represent a short-stalked member of
A. niveus.

In their most typical manifestation, the conidial heads of Aspergillus
niveus are loosely columnar; vesicles are dome-like, and fertile over the
upper one-half to two-thirds only. They thus stand out in sharp contrast
against the typically globose heads and completely fertile vesicles of A.
candidus. But all heads of A. candidus are not globose, and all vesicles are
not fertile over their entire surface. In most strains small columnar heads,
not particularly different from those of A. niveus, can be found (fig. 60 D).
Considering this character, together with (1) the snow-white heads, (2) the
smooth colorless conidiophores, and (3) the small smooth conidia of both
species, one can readily imagine that we are here dealing with two interlock-

204 A MANUAL OF THE ASPERGILLI

ing groups. One basic character separating the two is the production of
sclerotia. Characteristic reddish-purple to black sclerotia are commonly
found in white-spored strains producing wholly or in part large globose
heads (A . candidus) ; they have never been seen in white-spored strains pro-
ducing only loose columnar heads (A . niveus). Until additional intermediate
forms are isolated and studied, the relationship between these white-spored
forms must remain a matter of conjecture, although in this manual they
are placed adjacent in what we believe to represent a natural placement of
the different groups.

Pathogenesis

Strains of A. terreus grow under a wide range of temperature, including
37° C. It is not surprising, therefore, that an occasional member of the
series is reported as a human parasite. One of these was re-described as
Stcrigmatocystis hortai by Langeron (1922). This culture, NRRL No. 274
(Thorn No. 5071.1), received from France, was originally isolated from a
human ear in Brazil. It is believed to be type and represents a characteris-
tic strain of A. terreus. Another was found in metastatic lesions on a corn-
husker's hand and forearm in Nebraska. Recently a strain was isolated
from an aborted fetus from a cow in Maryland; the culture was entirely
typical of A. terreus.

Occurrence and Economic Importance

Members of the Aspergillus terreus group are typically soil organisms,
hence are most abundant in soil and upon decaying vegetation. They
frequently occur, however, upon a great variety of materials useful to man,
including grains in storage, straw and forage products, cotton and other
fibrous materials not adequately protected from excessive moisture, etc.
Aspergillus terreus and Aspergillus carneus are especially widespread in warm
arable soils, and have been isolated in great abundance from soils collected
in southern and southwestern United States. They are generally less com-
mon in forest than in cultivated soils, and are rarely found in acid forest
soils from the colder temperate zone. There is little evidence that these
forms are especially active agents of decay, but their great abundance in
nature indicates that they undoubtedly play a significant role in the slow
decomposition of organic materials. Aspergillus terreus and A. carneus
grow well at temperatures of 35 to 37° C, a character which possibly
accounts for their great abundance in southern soils and their relative scar-
city in soils from northern areas.

Aspergillus terreus has become of special biochemical interest since the
discovery in 1939 by Calam, Oxford, and Raistrick that certain strains of
this species are capable of producing itaconic acid from sugars. Extensive

Plate VI

A (upper left), Aspergillus Candidas Link, XRRL No. 308. B (upper right), Aspergillus niger group, strain
NRRL No. 67. C (center left), Tan-spored mutant of strain 67 produced bv ultraviolet radiation. D (center
"gD d' AsPer0illus arenaceous Smith, NRRL No. 517. E flower left ), Aspergillus alhaceus Thorn and Church.
XRRLN'o. 315. F (lower right), Aspergillus wentii Wehmer, NRRL No. 375. All cultures growing upon
Czapeks solution agar. (Color photographs by Haines, Northern Regional Research Laboratory. Repro-
duced through co-operation of Chas. Pfizer & Co., Inc.)

THE ASPERGILLUS TERREUS GROUP 205

investigations on this fermentation have been conducted in the Fermenta-
tion Division of the Northern Regional Research Laboratory, and papers
reporting these studies are currently in press. More than 300 strains have
been tested and the most productive cultures selected for intensive study.
Improved nutrient solutions have been developed and critical environmen-
tal factors explored with the result that substantial yields of itaconic acid
are now obtained. It is believed that this process may become of industrial
importance within a reasonable period of time. (See Lockwood, Raper,
Moyer, and Coghill; Lockwood and Reeves; Lockwood and Ward; Moyer
and Coghill; and Raper, Coghill, and Hollaender, in the Topical Bibliog-
raphy under "Itaconic Acid.")

Timonin (1942) reported the production of citrinin by a white-spored
Aspergillus identified by him as one of the A. candidus group. Careful
examination and comparison of his culture with representative strains of
A. candidus, A. carneus, and A. niveus show its true relationship to be with
A. niveus in the terreus group, as it is considered here, rather than with A.
candidus.

Chapter XVI
THE ASPERGILLUS CANDIDUS GROUP

Outstanding Characters

Conidial heads persistently white or becoming yellowish cream in age;

typically globose, but approaching columnar in small heads.
Conidiophores smooth, colorless or slightly yellowed in terminal areas.
Sterigmata in two series, with primaries often much enlarged, sometimes

varying greatly in size within the same head.
Conidia globose or subglobose, smooth.
Sclerotia present in some strains, dark, approaching purple to black

when mature.

Grouping by color lead Thorn and Church (1926) to establish their Section
IX, or the so-called "White-spored Aspergilli." Only vaguely did they
indicate that they had included a heterogeneous lot rather than described
a natural group. Further study of the strains included in the "white"
section showed the tardy development of colors approaching avellaneous
or even carneus. The continued comparison of great numbers of strains
in all groups has revealed such wide variations in color that the authors
have come to regard whiteness, or lack of color, as a character of secondary
importance in the allocation of strains to particular groups. This view-
point is supported by the appearance, under controlled conditions, of
white variants or mutants in a number of colored series. Yuill (1939)
observed and isolated such colorless mutants from Aspergillus fumigatus
and A. nidulans; Steinberg and Thom (1940) reported the same type of
mutant for the former species; while Raper, Coghill, and Hollaender (in
press) have succeeded in producing white mutants in Aspergillus terreus
by irradiating spores with ultra-violet. Colorless members of the Asper-
gillus glaucus group, represented by A. niveo-glaucus, have been isolated by
Blochwitz, Thom and Raper, and other investigators. Long before the
work of Yuill, Schiemann (1916) had developed two color variants of
Aspergillus niger, A. schiemanni (Schiemann) Thom (1926, p. 172)
and A. cinnamomeus Schiemann (1912) which differed only from the
parent strain by a progressive reduction of the amount of coloring
substance, presumably the aspergilline of Linossier (1891). Steinberg
and Thom (1940), working with a strain of A. niger, again produced
variants approximating those of Schiemann, and Whelden (1940) secured
the same by irradiating spores of A. niger with cathode rays. In cultures

206

THE ASPERGILLUS CANDIDUS GROUP 207

collected from nature from world-wide sources, strains characterized by
heads approaching white are occasionally observed in other groups. It is
apparent, then, that the capacity to produce a coloring substance, while
ordinarily inherited or passed on in successive colonies of an organism, is
not always uniformly maintained.

In the present treatment, the writers have sought to include within the
Aspergillus Candidas group only such forms as are clearly and closely related
to it. The group is thus limited, essentially, to a single series containing
only one clearly definable species, A. candidus. Different isolations vary
materially in their general cultural appearance and in the details of their
microscopic structure. Nevertheless, all possess the typically globose,
white to dull buff or light gray conidial heads, the smooth colorless coni-
diophores, and the small, smooth, colorless conidia.

To facilitate recognition of members of the Aspergillus candidus group
and to assist in the proper assignment of other white or light-colored
species and strains, a general key covering all of these forms is presented.

GENERAL KEY OF WHITE ASPERGILLI

A. Heads (large ones) globose or radiate; conidiophores smooth-walled, colorless or

yellowed toward the vesicle only; sclerotia occasionally seen A. candidus group

B. Heads white, hemispherical to columnar; conidiophores smooth-walled, colorless

A. niveus series, see p. 202

C. Heads initially white, tending to be columnar; conidiophores smooth-walled,

showing some shade of yellow; contorted hulle cells usually found

A. flavipes, series, see p. 179

D. Heads white, borne upon long, smooth-walled, colorless conidiophores terminat-

ing in clavate vesicles, sterigmata in two series

White-spored phase of A. janus, see p. 187

E. Strains of white Aspergilli possessing the basic characters of their colored counter-

parts also occur as mutations in the A. fumigatus, A. nidulans, A. terreus, and
glaucus groups.

Aspergillus candidus Link, Obs. p. 16, 1809. Thorn and Church, The

Aspergilli, p. 157. 1926.

Colonies upon Czapek's solution agar persistently white, or becoming
cream or yellowish-cream in age (PI. VI A and fig. 59), often thin, veg-
etative mycelium often largely submerged, surface growth usually con-
sisting of conidiophores and heads, and with scanty sterile mycelium or
anastomosing ropes of hyphae bearing short-stalked fruiting structures;
sclerotia produced in occasional strains; reverse usually uncolored. Heads
white, globose, radiate, varying in the same culture from large globose
masses 200 to 300m in diameter to small heads often less than 100/x in
diameter, commonly more or less elongated in heads with incomplete de-
velopment of sterigmatic surface. Conidiophores varying with the strain,

208

A MANUAL OF THE ASPERGILLI

in short or dwarf races less than 500m long, in other strains ranging up to
500 to 1000m or longer, varying from 5m in diameter in dwarf forms to 10
to 20m in long-stalked forms, with walls thick, smooth, colorless or slightly
yellowed near the vesicle in certain strains in age. Vesicles typically

Fig. 59. Aspergillus candidus. A, Strain XRRL No. 305 on Czapek's solution
agar at room temperature, 10 days. B, Strain NRRL No. 314 on Czapek's solution
agar at room temperature, three weeks. Note contrast between compact colony
of No. 305, consisting of crowded conidiophores, and loose spreading colony of No.
314 in which production of conidial heads is very irregular. C, Strain NRRL No.
312, portion of colony showing scattered black sclerotia, X 3. D, Strain NRRL
No. 308, conidial heads, X 18.

globose, ranging from 40m in diameter in very large heads (fig. 60 A), and
typically fertile over the whole surface, to small globose heads (fig. 60 B),
often very much reduced to support simple groups of sterigmata appearing
almost penicillate (fig. 60 D). Sterigmata typically in two series, usually
colorless, primary varying greatly in different strains, in different heads of

Fig. 60. Conidial structure of Aspergillus candidus, X 900: A, Large globose head
showing large primary sterigmata, strain NRRL No. 312; B, Smaller globose head
showing small primary sterigmata, strain NRRL No. 308; C, Head showing primary
sterigmata of variable size, some septate, strain NRRL No. 312; D, Diminutive
head, same strain. Great variation in dimension is characteristic of the fruiting
apparatus of most A. candidus strains.

209

210 A MANUAL OF THE ASPERGILLI

the same strain, and occasionally in the same head (fig. 60 C), ranging from
5/i in length in some cases to 15 or 20/t and even 30/x under other conditions,
commonly septate; secondary sterigmata usually uniform in all heads,
from 5 to 8/x by 2 to 2.5 or 3/x. Conidia colorless, globose or subglobose
in most strains to elliptical or barrel-form in others, thin-walled, 2.5 to
3.5ju or occasionally 4/x, smooth.

Reddish-purple to black sclerotia, consisting of thick-walled, parenchyma-
like cells, occur in many strains (fig. 59 C).

As there is no possibility of determining which of the white Aspergilli
was in Link's possession, the name is here used to cover a whole series of
strains which are found everywhere but are most frequent in the later
stages of decay in vegetation and are especially characteristic of moldy
grain. Included within the series as we consider it are two rather different
cultural entities. The first of these is characterized by thin colonies in
which the mycelium is largely submerged and only fruiting structures,
commonly arranged in concentric zones, rise above the level of the sub-
stratum (fig. 59 B). In other strains, colonies are rather floccose and some-
what felted and often attain a depth of 2 mm., with fruiting structures
arising from aerial as Well as submerged mycelia. Sclerotia, generally in
purple or black shades, regularly and consistently develop in many strains
including representatives of both of the above colony types. Conidiophores
vary greatly in size and characteristically reach their greatest dimensions
upon the drier portion of agar slants, or upon slightly moistened grain and
other comparable products low in water content. The sterigmata in
organisms of this group warrant particular attention since the two series
commonly, but not consistently, differ tremendously in size. In many
strains the primary sterigmata are characteristically wedge-shaped and
reach dimensions of 25 to 30m by 10 to 12/* (fig. 60 A and C) ; such structures
are commonly septate. In other heads from the same culture the primary
sterigmata may be relatively small and measure 6 to 8/x by 2.5 to 3. 5/i
(fig. 60 B). Secondary sterigmata are consistently small and of the dimen-
sions indicated in the species description.

"White" Aspergilli regularly constitute a normal element in the micro-
population from moist or improperly dried grains and of comparatively
dry vegetation undergoing slow decay. From such material, many in-
vestigators have described molds characterized by white heads which
obviously belong in this group, but without supplying sufficient critical
data to permit subsequent verification of the exact types under study.
Some of these descriptions were based upon molds growing in culture;
more of them were not. A few of the more tangible of these probable
synonyms will be briefly considered in this connection ; others will be found in
the general species index (pp. 331-?).

THE ASPERGILLUS CANDIDUS GROUP 211

S. alba Bainier (Bull. Soc. Bot. France 27: 30. 1880) was isolated from oatmeal,
and while incompletely described, obviously represents a member of the A. candidus
series. Several publications present elaborate comparative tables to separate strains
accepted as A. Candidas and A. alb us, but the many strains obtainable vary into each
other so completely that little or no basis for separation exists in fact.

A. albus Wilhelm (Beitr. z. Kenntn. d. Pilzgattung Aspergillus, Inag. Diss. Strass-
burg, p. 69, 1877) was described with characters which clearly ally it with A . candidus
but without sufficient differences to separate it from other members of this group.

S. blanc-jaune Bainier nomen nudum — A culture from Bainier's collection received
by Thom under this name (No. 4640.490) represents a somewhat diminutive but other-
wise typical member of this series.

S. albo-lutea Sartory and Meyer (Cited by Blochwitz in Ann. Mycol. 31: 73. 1933).
Conidia were reported as turning yellowish in age. This character is common to
many members of the group and has been so noted by Wehmer (1889-1901), Thom and
Church (1926), and others. Retention of the species name is not warranted.

A. basidiosepla Sartory, Sartory and Meyer (Ann. Mycol. 27: 317-320, PI. 7. 1929)
apparently represents a member of the A. candidus series with comparatively long
(28 to 30m) primary sterigmata which in age are characteristically septate. This char-
acter, which appears also in some members of the A. niger and A. ochraceus groups,
however, is not sufficiently unique to warrant specific separation.

A. niveus var. major Blochwitz (Ann. Mycol. 32(1/2): 86. 1934). Described as
showing vesicles globose, rarely oboval, or pear-shaped entirely covered with radiat-
ing sterigmata which are rarely absent toward the base; closely growing conidio-
phores 2 to 2.5 mm. high. These characters suggest relationship with A. candidus
rather than A. niveus.

A. okazakii Okazaki, in Centralb. f. Bakt. etc., 2 abt., 19, p. 481-484, taf. I. 1907;
see also Centralb. f. Bakt. etc., 2 abt., 42, p. 225. 1914. This is cited by Saccardo
in Syll. 22: 1260. 1913, as S. okazakii Saito but apparently without adequate ground
for attributing the name or description to Saito.

Colonies described as white to sulphur yellow; conidiophores hyaline, straight or
sinuate, smooth or asperulate, 200 to 500m by 8 to 12m figured as undulate, especially
toward the base, with walls 2 to 3m thick; heads 80 to 100m in diameter; vesicles 12 to
40m in diameter; primary sterigmata 15 to 20m by 6 to 8m, secondary 8 to 14m by 2.5 to
4m; conidia globose, hyaline, 2.5 to 5.4m, smooth, with connectives. In the event that
continued study of these white forms reveals the existence in nature of strains with
more or less roughened conidiophores and conidial heads ranging to yellows, recogni-
tion of A. okazakii as a separate species would be warranted. Based upon current
information, however, we believe it preferable to consider it synonymous with A.
candidus Link.

A. sachari of Chaudhuri and Sachar (Ann. Mycol. 32: 95. 1934) is more or less
arbitrarily left where the authors put it — as one of the A. sulphureus series near A.
quercinus in the A. ochraceus group. The heads are pale yellow, the sclerotia are
near the colors of that group, but the conidiophore is described as colorless and
smooth which would put it in A. candidus.

A. sterigmatophorus Saccardo, in Mycologicae Venetae Specimen. Atti d. Soc.
Ven. Trent, d. Sci. Nat. 2, fasc. 2: 232. Tab. XVII, fig. 5-8. 1873; Syn. S. italica
Sacc. in F. italici no. 109, 1881 ; changed to S. italica Sacc. as a note only in Michelia
1:91. 1877; Latin diagnosis of S. italica in Saccardo Sylloge 4: 72. 1886. Described
from decaying corn kernels (Zea mays): white, sparse, with conidiophores un-
branched, 2 to 3 septate above; with vesicles globose; sterigmata described as dicho-
tomously or trichotomously branched with ultimate cells bearing conidial chains;

212 A MANUAL OF THE ASPERGILLI

conidia globose about 6^ in diameter, with connectives. The description repeats
observations as to occurrence and appearance that are frequently seen. No one
has since reported a member of the A. candidus group with conidia 6m in diameter.
Whether the organism described by Saccardo was a large-spored mutant, not since
isolated, remains open to question.

OTHER WHITE ASPERGILLI

Other Aspergilli characterized by white heads but differing basically
in morphology from the A. candidus group occur in the Aspergillus glaucus,
A. nidulans, A. fumigatus, and A. terreus groups. Except for an absence
of spore color, these duplicate the morphology of the groups to which
assigned. In fact, in all cases except that of A. niveo-glaucus in the A.
glaucus group, they represent colorless mutations produced experimentally
from typical parent strains (Yuill, 1939; Steinberg and Thorn, 1940; Raper,
Coghill, and Hollaender, in press). While A. niveo-glaucus was isolated
from nature and hence its parentage is not known, it is suspected that
this represents a mutation of some form close to Aspergillus echinulatus.
A. halophilus of Sartory et al (Ann. Mycol. 28: (3/4) pp. 362-3, PI. 3, 1930)
similarly belongs in the A. glaucus group. Attention has been called earlier
to the fact that A. candidus differs from A. niger primarily in the absence
of color and in possessing smooth spores. The question may arise whether
we are not here dealing with a whole series of mutations from colored forms.
While this is possible, no proof is at hand. The fact that they constitute
such a typical and abundant element of the micropopulation of soil, decaying
vegetation, etc., demands that they be considered along with other major
groups of the Aspergilli quite aside from any questions of possible origin.

Sclerotia

In this arrangement of the Aspergilli, sclerotia, as compact globose or
subglobose bodies composed of thick-walled pseudo-parenchyma, are not
found in the groups characterized by the production of perithecia and
ascospores, and only rarely, if at all, in groups characterized by the presence
of hulle cells. In the great groups beginning with A. candidus, sclerotia
appear with sufficient frequency to be morphologically significant as
indicative of class relationship. Fundamentally, the typical A. candidus
strain differs little from the black Aspergilli except for the absence of the
dark color and rough spores.

Group Relationships

While there is much evidence of relationship with the black Aspergilli
(smooth-walled conidiophores, globose vesicles and heads, and the presence
of sclerotia), there are also certain indications of relationship to Asper-
gillus niveus. Typically both are characterized by snow-white conidial

THE ASPERGILLUS CANDIDUS GROUP 213

heads, and in all strains of Aspergillus candidus there are more or less abun-
dant small heads which bear few and loosely arranged sterigmata in a
manner strongly suggestive of typical conidial structures of A. niveus.
Although it is our belief that these similarities in structure do not of
necessity reflect close relationship between A. candidus and A. niveus, we
do feel that there is need for additional study of strains which appear to be
more or less transitional between the two groups.

Occurrence and Economic Importance

Members of the Aspergillus candidus group are very widely distributed
in nature and occur with reasonable frequency upon vegtation in the later
stages of decay. They are especially common upon moldy grains and
are obviously able to grow in the presence of a very limited amount of
moisture.

The biochemical and physiological activities of these form have not been
studied extensively. A. okazakii was employed by Okazaki (1907 and
1914) for the production of a proteolytic enzyme preparation, "digestin,"
and is the basis of a Japanese patent, No. 11461, covering this process.
Recently Timonin (1942) has employed a strain reported as belonging to
the A. candidus group for the production of citrinin. Upon examination,
however, this strain is found more nearly to represent A. niveus than A.
candidus in the sense it is considered here.

Chapter XVII
ASPERGILLUS NIGER GROUP

Outstanding Characters

Conidial bonds carbon black, brownish-black or purple-brown; in mutants,

shading toward colorless but not actually white.
Heads typically large and globose; but small heads produced in some

strains, in extreme cases consisting of only a few sterigmata and chains

of eonidia.
Conidiophores smooth, colorless or tinged with yellow --brown colors in

the upper one-third or less, splitting lengthwise into strips and shreds

when broken.
Vesicles globose in large heads, fertile over the entire surface; in small

heads often reduced to dome-like apices of short conidophores ("fumi-

gatifonn")-
Conidia rough, mostly showing bars or bands of brown-black coloring

matter.
Sclerotia characteristic of many strains, more or less irregular, ranging

from buff through gray to almost black.

The designation "Aspergillus niger"1 is commonly used to cover a great
aggregate of Aspergilli differing in details of morphology, but having in
common the production of conidial heads which are black, brownish-black,
purplish-brown, or in some strains lighter in color but retaining the general
appearance oi the group.

When examined by the hundred as they are isolated from natural sources,
the vast majority of cultures studied show the general morphology of
Aspergillus niger van Tieghem. if a reasonable allowance be made for strain
variation which is characteristic of all of these groups of cosmopolitan molds.
At the same time, other organisms scarcely distinguishable in general
cultural appearance often show marked differences in microscopic char-
acters, hence have formed the basis of species descriptions. Other species
have been segregated by various authors in the belief that they bore an
obligate relation to the particular substrata from which isolated (e.g..
.4. strychni. A. ficuum), but such specificity has not proved dependable.
There are. in addition, a considerable number of probable variants whose

1 Biourge in his last manuscript proposed the use of the name Pulli for the black
Aspergilli in recognition of the belief that Mieheli J~29^ had one of them before him
as his "Aspergillus capitolus capitulo pullo."

214

ASPERGILLUS NIGER GROUP 215

origin is not known, and which may or may not have genetic connection,
but which do show superficial resemblances a1 least. Some of these have
been described as species and can be more or less readily identified. Al-
together, there is a considerable Lisl of names which, at one time or another,
have been applied to the black Aspergilli. It is now quite impossible to
interpret many of these descriptions, or to find out exactly what type of
organisms the describers had under observation. Realizing the futility
of attempting to separate all of these species, we have endeavored to in-
clude in the group key only such forms as possess well-marked character-,
and those which the mycologist may reasonably expect to encounter in
laboratory culture.

Group Key

I. Sterigmata in two scries.

A. Conidia mostly less than 5m in diameter Aspergillus niger series

1. Conidia strongly colored, rough, definitely marked with echinulae,

tubercles or color bars.

a. Primary sterigmata mostly under 20m in length.

1/ Colonies with a penetrating actinomyces-like odor

A.foetidus n. sp. <= A. aureus Xak.)
2.' Colonies without such odor.

a. Colonies black or deep brown to black.

1." Heads abundant over the entire colony

A. awamori Xakazawa
2." Heads few, scattered at margins of colonies

A. miyakrensis Xakazawa
b.' Colonies yellow-brown

Occasional Isolates. See also A. wentii group.

b. Primary sterigmata mostly 20 to 30m long

A. niger van Tieghem series and species.

c. Primary sterigmata mostly 40 to 60m long

A. phcenicis (Cda.j Thorn

d. Primary sterigmata up to 100 to 120m long

A. pulverulentus (McAlpinej Thom

2. Conidia with color almost suppressed or diffused, leaving smooth walls

with colorless spinules.

a. Conidia almost colorless, in mass very pale cimmamon

A. niger mut. cinnamomeus n. comb.

b. Conidia with color diffused, sometimes a trace of roughening

A. niger mut. Schiemanni n. comb.

B. Conidia more than 5m in diameter Aspergillus carbonarius series

1. Primary sterigmata less than 20m in length.

a. Conidia purplish-black, 6 to 10m in diameter

A . atropurpureus Zimmerman

b. Conidia brown, 5 to 7 or 8m in diameter. .A. fumaricus Wehmer

2. Primary sterigmata 20 to 45m in length. Conidia 6 to 8m in diameter

A.fonsecaeus n. sp. (= S.fusca Bainierj

3. Primary sterigmata up to 100 or 120m. Conidia 5.5 to 10.5m

A. carbonari u.s_ fBainier) Thom

216 A MANUAL OF THE ASPERGILLI

II. Sterigmata in one series (Secondary sterigmata occasional in some strains)

Aspergillus luchuensis series

A. Colonies black or black-brown.

1. Sterigmata 6 by 3/* (very short) A. luchuensis Inui

2. Sterigmata about 15 to 20/xconidia 3 to 3.5yu.. .A. nanus Montagne

and/or A. subfuscus Johan-Olsen

B. Colonies in reddish-brown shades.

1. Conidia globose A. japonicus Saito

2. Conidia elliptical A. violaceo-fuscus Gasperini

Because of their great abundance in nature, the series most closely-
related to and including van Tieghem's species, based upon strains which
satisfy his original description in a somewhat broadened sense, will be
discussed first.

ASPERGILLUS NIGER SERIES

Species Characterized by Comparatively Small Primary Sterigmata and

Small Conidia

Aspergillus niger van Tieghem, in Ann. Sci. Nat. Bot., s. 5, t. 8, p. 240.

1867.

Synonym: Sterigmatocystsis nigra van Tieghem, in Bui. Soc. Bot.
France 24: 102-103. 1877. See also Thorn and Currie, Jour. Agr.
Res. 7: 1-15. 1916; and Thorn and Church, The Aspergilli, p. 167.
1926.

Characterization: Colonies rapidly growing with abundant submerged
mycelium, colorless, or in some strains with more or less yellow color in
the hyphae and in the substratum, with aerial hyphae usually scantily
produced, but abundant in age in certain strains. Conidial heads fuscous,
blackish-brown, purple-brown, in every shade to carbonaceous black
(PI. VI B), varying in intensity with the quantity of coloring matter pro-
duced; typically globose or radiate (fig. 63 C), commonly up to 300, 500,
or occasionally 1000m in diameter with periphery variously splitting into
radiating columns of conidia; small heads, more or less columnar and
consisting of a few conidial chains often borne on trailing hyphae or short
conidiophores near the substratum. Conidiophores mostly rising directly
from the substratum, uncolored or yellow to brown near the vesicle only,
smooth, with walls thick, frequently uneven on the inner surface and split-
ting lengthwise into strips when broken (fig. 64 B and C), unseptate or
with occasional thin septa, varying greatly in length and diameter in
different strains and in colonies on different media or even in sections of
the same colony, thus ranging from strains with conidiophores 200 to 400m
by 7 to 10m to forms with conidiophores several millimeters long and 20m
or more in diameter. Vesicles globose or subglobose, thick- walled, com-

Fig. 61. Aspergillus niger group: cultures growing upon Czapek's solution agar
at room temperature, 10 days. A, A. niger, NRRL No. 334, typical strain. B, A.
niger mut. schiemanni characterized by colonies light brown in color. C, A.foetidus,
NRRL No. 341, characterized by a yellowish vegetative mycelium and a strong
actinomyces-like odor. D, A . niger, NRRL No. 346, characterized by the production
of abundant sclerotia. E, A. phoenicis, NRRL No. 1956, characterized by long
uncrowded conidiophores. F, A. riolaceo-fuscus, NRRL No. 360, characterized by
compact, close-textured colonies and small heads with uniseriate sterigmata.

217

Fig. 62. Conidial structures in the Aspergillus niger group, X 500: A, A. niger,
NRRL No. 326, typical head showing globose vesicle and primary sterigmata about
twice the length of secondaries; B, A. phoenicis, NRRL No. 1956, characterized by
long and occasionally septate primary sterigmata; C, A. carbonarius, NRRL No.
369, characterized by la^rge primary sterigmata and large conidia; D, A. violaceo-
fuscus, NRRL No. 360, characterized by small heads and sterigmata in a single series.

218

ASPERGILLUS NIGER GROUP 219

monly 20 to 50m, occasionally up to 100m in diameter (fig. 62 A), colorless
or more commonly more or less intensely yellow-brown. Sterigmata in
one series in young colonies and in small heads, but typically in two series,
colorless at times, usually more or less intensely brown, even carbonaceous,
primary sterigmata closely packed, covering the vesicle, varying greatly
in size in the same colony but usually 20 to 30m in length by 6 to 8m in
diameter at the outer end; secondary sterigmata more uniform, ranging
usually from 6 to 10m by 2 to 3m (fig. 62 A), both series often more or less
brown to almost black. Conidia globose when ripe, with walls at first
smooth with diffused brown or fuscous color (Ridgway, PL XLVI), then
rough or spinulose from coloring substance deposited as tubercles, bars
or loops between the outer primary wall and the inner, or secondary, wall,
mostly 2.5 to 4m, occasionally up to 5m in diameter.

Sclerotia globose, superficial, regularly produced by certain strains
(fig. 61 D), sporadically by some, and not found in many others.

A. niger approximating the description of van Tieghem furnishes the
most common morphological entity among the black Aspergilli. A few
of the substrata and locations found in our record include chronic irritants
in the human ear, pin-point colonies in the human lung, spoiling raw sugar,
rancid butter and other fats, floating and submerged mycelium in many
chemical solutions. It is abundant in soil cultures from every part of
the world, and apparently especially so in the tropics. Molds under this
name have been used in literally hundreds of biochemical investigations.

The introduction of a complete description from culture for each member
of the series typically represented by van Tieghem's A. niger, but which
vary from it in detailed measurements, calls for repetition of many common
characters. In place of such descriptions the names and citations of the
forms selected, either as unique or as representing sections of the series
often encountered, are presented with the more important differences
which furnish the bases of separation.

Aspergillus foetidus n. sp.

Synonym:^, aureus Nakazawa, in Inst. Gov't. Res. Formosa, Rept.

Vol. 1, 1907.
Not:i4. aureus Berkeley, in English Flora Vol. 5, p. 346. 1836.
Not S. aurea Greco, in Origine des Tumeurs et Mycoses

Argentines, Buenos Aires, pp. 671-694, fig. 418-428. 1916.

Colonies upon Czapek's solution agar rather slow growing, producing a
floccose basal mat of mycelium with abundant but uncrowded black heads
above a mass of pale orange mycelium which is deeply orange in reverse.
Heads up to 225m in diameter are borne on conidiophores about 500m

220 A MANUAL OF THE ASPERGILLI

long; vesicles comonly 20 to 30m in diameter, occasionally much larger.
Primary and secondary sterigmata are both 7 to 10m by 2 to 4m; conidia
are globose, spinulose, up to 4 or 4.5m-

Colonies have a penetrating actinomyces-like odor (also like Penicillium
biforme noted in Thorn, The Penicillia, p. 320. 1929), unlike any other
species of Aspergillus. Numerous experiments over several years failed
to justify the belief that the culture was contaminated with some actino-
mycete. The species is known only from Nakazawa's isolates which have
maintained the odor and orange color for many years.

In the Awamori fermentation, A. foetidus (A. aureus Nakazawa) was
j8, the unfavorable organism which gave a yellow color to the "Koji" used.
Nakazawa, Simo, and Watanabe (Jour. Agr. Chem. Soc, Japan, No. 144,
pp. 931-974, illustr., 1936) listed five varieties of A. aureus as follows:
var. minor, var. murinus, var. acidus, var. pallidus, and var. brevis.

Aspergillus awamori Nakazawa, in Inst, of Gov't. Res. Formosa, Rept.

Vol. 1, 1907 and Vol. 2, 1912.

The measurements given are only slightly different from those of A.
foetidus ( = A. aureus). The unique odor and the yellow color in the
mycelium and substratum were lacking. Colonies blackish-brown (deep
chocolate) when heads were fully developed, conidiophores 1 to 2.5 mm.
by 9 to 15m; vesicles globose, 30 to 45m hi diameter; primary sterigmata
9 to 12m by 3.5 to 5.5m, and secondary 4.5 to 8m by 1.5 to 3.5m; conidia
globose or somewhat elliptical 3 to 5m in long axis, fairly spinulose.

Nakazawa, Simo, and Watanabe (Jour. Agr. Chem. Soc. Japan, No. 144,
pp. 931-974, illust., 1936) studying the fermentation industries of Formosa
found two general types of Aspergilli in the Awamori fermentation, a and
(3. Type a was A . awamori for which they described the following varieties :
var. minimus, var. piceus, var. ferrugineus, var. fuscus, var. fumeus. This
"awamori" series produced the more desirable type of product; citric acid
was present, as well as alcohol due to the yeast used in the inoculum.

Aspergillus niger var. fermentari us Nakazawa, Simo and Watanabe, in Jour. Agr.
Chem. Soc. Japan, 10(2) 1934. pp. 171-172, summarized on p. 184. Reported as a
variety with "conidiophores 1037 to 2438m by 13.1 to 16.0m; vesicles 22.6 to 73.6m; pri-
mary sterigmata 12.7 to 16.1m by 3.3 to 7.2m, secondary 6.8 to 9.8m by 3.3 to 4.6m; and
conidia globose 2.3 to 4.6m in diameter." This form obviously belongs close to A.
awamori.

Aspergillus miyakoensis Nakazawa, Simo, and Watanabe, in Agr. Chem.
Soc. Japan, Jour. 12(9): 963-4, fig. on 973. 1936.

The colonies figured by the authors show a cottony mycelium with
long-stalked heads in a broad zone near the margin. The measurements

ASPERGILLUS NIGER GROUP

221

differ from ^4. foetidus as follows: primary sterigmata are reported to be
12 to 20/x by 4.4 to 9m in contrast to secondaries 7 to 10/x by 2.5 to 5/x, whereas

Fig. 63. Conidia, Aspergillus niger group. A , Single long chain of conidia, X 1000
(Photograph by Edward Yuill). B, Conidia of a typical strain of A. niger, NRRL
No. 344, X 700. C, Conidia of the citric and gluconic acid producing strain, NRRL
No. 67, X 700. The large size and coarsely roughened walls are characteristic of
the conidia of the latter strain.

in A. foetidus both series measure 7 to 10/x by 2 to 4m; conidia are globose
3.7 to 5.6m in diameter.

The species repeats the colony appearance of certain mutants produced
by means of chemical stimulants by Thorn and Steinberg (1939) from the
latter 's standard strain of A. niger.

222

A MANUAL OF THE ASPERGILLI

.4. hennebergi Blochwitz, in Ann. Mycol. 33: 238-9. 1935.

Colonies described as showing the colors and general aspect of ,4. tamarii or A.
wentii but with conidiophores browned as in the upper part of the conidiophores of
the A. niger group, and with "red" sclerotia; relationship doubtful. See also the
A. wentii group.

Aspergillus niger van Tieghem, in Ann. Sci. Nat. Bot., s. 5, t. 8, p. 240. 1867.

Van Tieghem 's strain is not fully verifiable among organisms now main-
tained in culture, although Biourge (personal communication) believed he
had it. We believe the name can be most appropriately used to cover

1

4, < v •*> t

A

t

B

Fig. 64. Broken conidiophores of Aspergillus niger, NRRL No. 326, X 700. A,
Conidiophore showing a clean break suggesting a glass tube. B, Broken conidiophore
showing fibrous wall structure. C, Portion of conidiophore crushed and further
revealing the fibrous structure of the wall.

the exceedingly abundant isolates having the approximate measurements
of sterigmata and conidia noted in van Tieghem's description. The reader
can assume, therefore, that in the opinion of the authors any possible
description for the species itself would read essentially like that already
given for the series on pages 216-219.

Species Characterized by Large Primary Sterigmata and Small Conidia

4 . phoenicis (Corda) Thorn, in The Aspergilli p. 175. 1926. In describing
the black Aspergillus found upon dates, Patouillard and Delacroix (Bui.
Soc. Myc. France 7: 118-120. 1891) compared their material to specimens

ASPERGILLUS NIGER GROUP 223

in the museum labeled "Ustilago phoenicis" and attributed to Corda, thus
establishing the identity of the organism of Corda, which was not recogniz-
able from any previous references. A. ustilago Beck (1892) is described
with the same measurements. The measurements of sterigmata place
A. phoenicis in the section of the group with primary sterigmata 50 to 60m
in length; it was first described as Ustilago phoenicis by Corda (Icones
Fung. IV., p. 9, pi. 3, fig. 26. 1840) and transferred by Patouillard and
Delacroix, as noted above, to 8. phoenicis (Corda) Patouill. and Delacr.
If we accept the use of "phoenicis" attributed to Corda as correct, this
species becomes the type of the section of the black Aspergilli with primary
sterigmata of intermediate length and conidia not over 4m in diameter
(fig. 61 E and 62 B). This was cited by Thorn and Currie as A. phoenicis
(Corda) Pat, and Delacr., and continued recognition of the species, to
cover a group of strains occasionally encountered, appears warranted.

Aspergillus pulverulentus (McAlpine) Thorn, in Jour. Agr. Res. 7:10-11.

1916.

Synonym: £. pulverulenta McAlpine, in Agr. Gaz. X. S. Wales (1896)
7: 302. 1897. See also Thorn and Church, The Aspergilli,
p. 179. 1926.

McAlpine's data include: "White to dirty yellow mycelium;" heads
155 to 340m in diameter, radiate with chains mostly separate; conidiophores
erect, stiff, up to 7 mm. by 20m with walls up to 5m thick; vesicles 70 to 170m
in diameter, globose or nearly so; primary sterigmata up to 144m long by
8m, secondary 14 to 18m long; conidia globose, rough, about 4m in diameter.
Colonies with these general characters have been studied in culture at
least twice and maintained for long periods; one came from Spain, the
other from Texas. A. strychni Lindau (Hedwigia Bd. 43, Rept, 5, p.
306-7. 1904) is one of this series.

Light Colored Forms

Aspergillus niger mut. cinnamomcus (Schiemann) n. comb.

Synonym: A. cinnamomeus Schiem., in Ztschr. Induktive Abstam. u.
Vererbungslehre, Bd. 8, Heft 112, pp. 1-35, 16 fig., 2 pi.
(1 col.) 1912. See also Thom and Church, The Aspergilli,
p. 164. 1926.

Colonies upon Czapek's solution agar at room temperature, rapidly
growing and spreading, producing an aerial growth of conidiophores and
heads reaching a pale cinnamon upon maturity. Reverse only slightly

224 A MANUAL OF THE ASPERGILLI

colored in the same shade. Conidial heads not crowded, globose. Conidio-
phores smooth, thick-walled, with upper portion more or less brown, about
1.5 mm. in length by 12 to 20m in diameter. Vesicles up to 40 to 50 m in
diameter, crushing readily. Sterigmata in two series; primary about
15 to 20m by 3 to 5m, sometimes larger, secondary about 8 by 2 to 3/jl.
Conidia 3 to 4m, thin-walled, globose or subglobose, smooth or nearly so,
almost colorless when viewed singly, pale yellowish to cinnamon in mass.
Diagnosis based upon culture NRRL No. 348 (Thorn No. 3534b) re-
ceived from . Schiemann as a mutation induced by introducing potassium
bichromate into the culture medium. Approximately the same mutant
appeared in Steinberg and Thorn's series of induced mutations (1939, 1940).
Occasional cultures close to A. cinnamomeus have ben obtained from un-
known sources in nature.

Aspergillus niger mut. Schiemanni (Schiemann) n. comb.

Synonyms: A. Schiemanni (Schiemann) Thom, Jour. Agr. Res. 7: 13.
1916.
A. fuscus Schiemann, in Ztschr. Induktive Abstam, u.
Vererbungslehre, Bd. 8, Heft Y2, p. 1-35, 16 fig. 2 pi.
(1 col.) 1912.
Not A. fuscus Bonorden (Bot. Ztg. Jahr. 19: 202. 1861);
Not S. fusca Bainier (Bui. Soc. Bot. France 27: 29, PI. 1, fig. 5.
1880)

Colonies upon Czapek's solution agar at room temperature, rapidly
growing and spreading, developing a surface growth of conidiophores and
heads forming a crowded fruiting area 2 to 3 mm. deep in slanted tubes,
becoming a shade of brown near fawn color (Ridgway, PI. XL); reverse
yellowish (fig. 61 D). Conidial heads large, fairly crowded. Conidiophores
coarse, 2.5 mm. or more long by 15 to 25m wide. Vesicles up to 50 to 60m
in diameter. Sterigmata in two series; primary, 15 to 40m by 4 to 6m,
sometimes larger, secondary 7 to 8m by 2 to 3m- Conidia thin-walled, smooth
except sometimes a trace of markings, 3.5 to 4.5 or 5m in diameter.

Culture NRRL No. 361 (Thom No. 3534C) was received from Schiemann.
Culture NRRL No. 362 was received from Biourge under the same name but
shows a much deeper brown color (near Natal brown, Ridgway, PI. XL).

The mutant, A. niger mut. Schiemanni, is distinguished from the parent
type of A. niger by the color and smoothness of its spores. The name,
A . fuscus Schiemann, is invalidated by the prior usage of the names A . fuscus
by Bonorden and S. fusca by Bainier.

Mutations of approximately this same type have been secured from
cultures of strain NRRL No. 67 (PL VI C) by ultra-violet irradiation

ASPERGILLUS NIGER GROUP 225

(Raper, Coghill, and Hollaender) ; from cultures of other black Aspergilli
through Cathode ray irradiation (Whelden 1939); by cultivation in the
presence of various chemicals (Steinberg and Thom: 1939, 1940); and
finally by the appearance in plate culture under normal conditions of a light-
spored sector in an apparently typical culture of Aspergillus niger. See
discussion Chapter VI. Two isolates labeled A. awamori Nakazawa
which resemble this mutant in color have been received from Japan, one
from Hanzawa, the other from Nakazawa. Both produce primary ster-
igmata less than 20m in length but otherwise are close to Schiemann's
mutant.

Probable Synonyms

Many species have been described by investigators working at different
periods and at widely separated stations which obviously belong within
the Aspergillus niger series as it is here considered. Some of these will be
briefly noted since they were reported to present unique cultural or morpho-
ological features, or since they account for interesting or important bio-
chemical reactions.

A. giganteus Mattlet (Ann. Soc. Beige Med. Trop. 6: 36. 1926) was described as a
new species with conidiophores 2 to 6 mm. long; conidia 4 to 5m in diameter, rough,
black-brown; but without other marks of separation. It would seem to have been a
rather extreme variant in the two characters.

A. atropurpureus Blochwitz (Ann. Mycol. 32(1/2) : 86. 1934.) Not A. atropurpu-
reus Zimmermann (Centralb. f. Bakt. etc., 2 Abt., 8, No. 5/7, p. 218. 1902). Bloch-
witz proposed to use the name A. atropurpureus for the purple-brown members of
the A. niger group as he obtained them from the tropics. He gave conidial measure-
ments as 2.5 to 3.5m in diameter, and added that chemical differences in the coloring
substances warranted separation from the black, or blacker forms.

A.ficuum (Reich) Hennings, in Hedwigia, 34, p. 86. 1895; and Reichardt, in
Verhandl. K.K. Zoll. Bot. Gesell. Wien, 17: 335, 1867. Regarded as A. niger by
Wehmer, in Centralb. f. Bakt. etc., 2 Abt., 18, No. 13/15, p. 394-395. 1907. See also
Thom and Church, The Aspergilli, p. 174. 1926.

Slight differences in morphology between this and A. niger v. Tieg. are reported by
Hennings, but disregarded by Wehmer. Culture NRRL No. 364 (Thom. No. 142),
received in 1909 from Westerdijk under this name, was reported by Thom and Currie
(1916) as the most rapid producer of oxalic acid of all strains of A . niger tested. There
is no morphological basis for separating this strain from A. niger, and it is distin-
guished in culture only by the tendency, after many years in laboratory culture, to
produce rather floccose colonies.

A.batatae Saito, in Centralb. f. Bakt. etc., 2 Abt., 18, No. 1/3, p. 34. 1907. Saito's
organism as described is close to A. niger: colonies reach brownish-black through
yellow shades; conidiophores 2 to 4 mm. by 12 to 20m, smooth, thick-walled, brown in
upper portion; vesicles 35 to 50m, globose; primary sterigmata 24 to 40m long by 8m
at apex, secondary 10 by 3.2m; conidia globose, brown, finely roughened, 4 to 5m in
diameter. A culture under this name from Formosa (NRRL No. 363) presented the

226 A MANUAL OF THE ASPERGILLI

comparatively large conidia described by Saito but possessed no other marks which
would assist in identification.

.4. iuteo-niger (Lutz) Thorn and Church, in The Aspergilli, p. 166. 1926. Syn.:
S. luteo-nigra Lutz, in Bull. Soc. Bot. France 53: 48-52. 1907. Thorn and Church
(1926) found one or more black Aspergilli in which the conidia appeared smooth in
ordinary laboratory slide amounts in which they were treated with alcohol followed
by lacto-phenol or other mounting fluids. Fragments floating in the mounting
medium led to the test of such conidia in dry mounts, in oil, and in pure glycerine.
Under these conditions, the conidia showed the typical marks of the A. niger series.
Since smooth conidia were the sole definite contrast between A. Iuteo-niger Lutz and
.4. niger v. Tiegh., the strains studied were accepted as Lutz' organism. Since that
time, it has been shown that many black Aspergilli, ranging widely among the variant
forms encountered, pass through a physiological stage in which the outer wall and
color bars, when subjected to alcohol or other fluid causing active osmotic currents,
break in pieces and float away leaving the conidia smooth and colorless, or only partly
shaded toward the dark color of the group. It thus appears that A. Iuteo-niger may
be dropped as failing to designate any definite group of strains and as failing to pre-
sent any character of diagnostic importance.

ASPERGILLUS CARBONARIUS SERIES

Species Characterized by Conidia in Excess of 5. 0^— Sterigmata in Two

Series

Aspergillus atropurpureus Zimmermann, in Centralb. f. Bakt., etc., 2 Abt.,

8, Xo. 5/7, p. 218. 1902.

Conidiophores hyaline or somewhat brownish in age, up to 800 by 16m
to 20/x ; vesicles 60 to 80m in diameter, hyaline to brown; sterigmata, primary
16 by 6m, secondary 3 to 4m by 1.5 to 2.0m; conidia globose, rough, with
prominent warts, purplish-black, 6 to 10m in diameter. Isolated from
Coffea liberica, in Java. Culture not studied by us. The species is possibly
valid, and is presented as representative of occasional forms characterized
by small sterigmata and large purple-black spores.

Aspergillus fumaricus Wehmer, in Ber. Deut. Chem. Gesell. 51, Xo. 14:

1663-1668, figs. 1-6. 1918.

This species was named, but not fully described by Wehmer, and was
not distributed by him. Its biochemical activity as a producer of fumaric
acid is covered in the paper cited above, together with the admission that
it belonged to the .4. niger group. Culture Xo. 4668.2 (C. Thorn) received
from Xeuberg under this name presents a variant strain of the group which
may be described as follows: Colonies producing a mass of yellow my-
celium; conidiophores scantily and tardily produced up to 1, 2, or 3 mm.
long, by 20 to 22m in diameter, smooth, bearing large radiate, yellow-brown
heads. Vesicles up to 60 or even 100m in diameter, with walls thin, easily
crushed. Sterigmata in two series, primary sterigmata up to 15m by 4 to

ASPERGILLUS NIGER GROUP 227

5m, secondary sterigmata rather coarse, some growing out into aborted
hyphae. Conidia commonly 5m, occasionally 7 to 8m in diameter, with
lengthwise color bars as in A. niger but paler in color.

Culture No. 4668.2 (Thorn) is accepted as correctly named. By de-
scription, A. fumaricus Wehmer differs little from A. atropurpureus Zimm.
except for the production of yellow-brown rather than purple-black conidial
heads. Differences in color definition and discrimination by different
workers tend to leave both species in doubt.

Aspergillus fonsccaeus n. sp.

Synonym: S. fusca Bainier, in Bui. Soc. Bot. France 27: 29, PI. 1, fig. 5;
1880. (Bainier's material grown upon moist bread in Tou-
louse in 1880, was preserved in Roumeguere's Fungi Gallici
Exsiccati Xo. 995)

Bainier described S. fusca in terms closely parallel to van Tieghem's
A. niger, except that the conidia were about double that species in size;
Bainier reported these as rarely exceeding 9.4m in diameter, while our exam-
ination of the exsiccati showed them to be mostly 5 to 8.5m in diameter
and marked as in A. niger and A. earbonarius. Da Fonseca, in Rio de Ja-
neiro, contributed a culture (Thorn Xo. 4707.878, XRRL Xo. 67, and strain
Xo. 67 of Herrick, May, and associates) possessing approximately these
spore measurements, which has retained its characteristic features for more
than 20 years and which has proved industrially useful. We believe,
therefore, that recognition of a species with sterigmata of intermediate
length and conidia about double the dimensions of A. niger van Tieghem
is warranted. The name A. fonsecaeus is proposed since the binomial
A. fuscus had already been used for an Aspergillus by Bonorden in 1861
(Bot. Ztg. Jahrg. 19: Xo. 29, p. 202) and has been used at least twice
subsequent to this. The species would then include strains characterized
by large, subglobose, coarsely roughened conidia ranging from 5.5 or 6.0m
to 8.5 or 9.0m (fig. 63 C), including the strain "67" used by Herrick, May,
AVells, Moyer, et al. of the Industrial Farm Products Research Division,
U. S. Department of Agriculture, Arlington Farm, Virginia, for the pro-
duction of citric and gluconic acids (see literature citations pp. 290-295).
The following description is based primarily upon strain XRRL Xo. 67:
Colonies upon Czapek's solution agar growing rapidly at temperatures
from 24° to 30° C, attaining a diameter of 7 to 8 cm. in eight to ten days,
consisting of a basal vegetative mycelium that is largely submerged and
colorless, and abundant conidial structures commonly arranged in more or
less conspicuous concentric zones; heads carbon black or brownish-black,
imparting to the colony a like coloration ; reverse colorless in young colonies,

228 A MANUAL OF THE ASPERGILLI

commonly darkening in age and often becoming almost black after 2 to 3
weeks. Conidial heads large, globose, radiate or with chains of conidia
massed in an indefinite number of loose divergent columns, commonly 300
to 500^ but often attaining a diameter up to 1 mm. Conidiophores varying
in length from 1.5 to 3.5 mm. but averaging about 2.0 to 3.0 mm., mostly
20 to 30m in diameter with walls 2.0 to 3.0m thick, smooth, often colored
in dark shades in the region beneath the vesicle. Vesicles globose, fertile
over the entire surface, commonly 50 to 75m in diameter (fig. 62 C), usually
in brown shades, often quite dark. Sterigmata in two series, usually
brown, often dark: primary sterigmata variable in different heads and in
different cultures, ranging from 15 to 20m by 6 to 8m in some to 35 to 45m
by 10 to 13m in others; secondary sterigmata ranging from 8 to 14m by 5
to 6.5m but averaging about 9 to 10m by 5 to 6m. Conidia large (fig. 63 C),
globose, conspicuously roughened with prominent color bars, ranging from
5.5 to 8.5m-

Since strain "67" appears in the industrial fermentation literature as
Aspergillus niger and has been consistently distributed under this name
over a period of several years, it is not our purpose here to challenge this
designation, for this binomial is often used in a very general sense to cover
any black member of this group. We do wish to emphasize, however, that
this strain does not represent the common type of black Aspergillus usually
isolated in routine examination of soil and moldy materials in general.
The fact that it possesses large spores is of the greatest value in checking
its purity and further commends it for use in industrial operations. This
strain was originally received from Rio de Janeiro, and recently Dr.
Dorival M. Cardoso of Sao Paulo, Brazil, has reported (personal communica-
tion) that he has repeatedly isolated large-spored forms apparently closely
related to it. It would, therefore, appear likely that this large-spored form
represents a type of organism much more abundant in South America than
in this country.

Aspergillus pulchellus (Speg.) Thorn and Church, in The Aspergilli, p. 181.

1926.

Synonym: Aspergillopsis pulchellus Speggazzini, in Myc. Arg. V. in Ann.
Mus. Nac. Buenos Aires Ser. 3, t. 13: 436. 1911.

This species was described with colonies intensely black; conidiophores 1 to 2 mm.
by 18 to 20m, with walls darkened; vesicles 50 to 60m in diameter; primary sterigmata
10 by 30m; and conidia 8 to 10m in diameter and rough. The species appears to differ
from van Tieghem's A. niger principally in its very large conidia, and it is extremely
doubtful if it could be separated from A. fonsecaeus as described above.

A. dipus of Ferdinandsen and Wing (Bot. Tids. 30: 220, fig. 6. 1910) represents
another organism undoubtedly close to the forms under consideration. The presence
of conspicuous foot cells, upon which character the species was based, is common to
all members of the group, hence, is not a valid basis for separation.

ASPERGILLUS NIGER GROUP 229

Aspergillus carbonarius (Bainier) Thorn, in Jour. Agr. Res. 7: 12. 1916.

Synonym : S. carbonaria Bainier, in Bui. Soc. Bot, France 27 : 27-28. 1880.

Colonies grown upon Czapek's solution agar show vegetative mycelium
white or with some yellow in submerged areas, broadly spreading, more or
less zonate; sclerotia produced upon the surface of the substratum in old
cultures; fruiting areas carbon-black. Conidiophores colorless below,
yellow to yellow-brown toward the apex, 4 to 6 mm. or more in length
and up to 25m hi diameter, with walls smooth, sometimes as much as 4m
in thickness. Heads globose, varying in diameter up to 500m- Vesicles
up to 90m in diameter, fertile over the entire surface, commonly with contents
yellow-brown to black and in old heads f orming with the primary sterigmata
a hard, brittle, carbonaceous mass. Sterigmata in two series, primary
sometimes one septate, from 20 to 40m long in young or small heads, and
up to 120m long in large heads by 5 to 13m in diameter at the apex, secondary,
8 to 14m by 3 to 6m- Conidia at first smooth, becoming rough when ripe,
5.5 to 10.5m in diameter. Colonies grow well upon all culture media used,
with temperature optimum below 37° C. A culture from Dr. A. F. Blakeslee
(NRRL No. 369, Thorn No. 4030.1) reproduces in detail the morphology
recorded by Bainier. A. carbonarius has also been received in culture from
the Gold Coast of Africa.

The same morphology was also found in one of Dr. Farlow's specimens,
S. acini-uvae Caballero (Bol. R. Soc. Esp. Hist. Nat, 28: 429. 1928).
This was described as it appeared upon rotting grapes as follows : vesicles
75 to 105m by 73 to 98m; primary sterigmata 59 to 80m by 13 to 22m and
secondary 15 to 20m by 5 to 7m ; conidia globose, rough, 6 to 10m- Blochwitz
in "Die Aspergillaceen" (Ann. Mycol. 27(3/4): 204, 222, and 232. 1929),
with part of the type specimen before him, declares it to be A. carbonarius,
apparently without cultivating it. Mosseray (LaCellule 43 : 222. 1934)
places Caballero's organism in Aspergillus pulchellus.

We cannot agree with Blochwitz (idem. p. 221, 222, fig. 12) in describing
the vesicle in A. carbonarius as subglobose, and figuring the head as hemi-
spherical. Bainier's original figures show the vesicle fertile to the very
base. This is characteristic of the strains observed from various sources.

ASPERGILLUS LUCHUENSIS SERIES

Species Normally Showing One Series of Sterigmata, Occasionally Two.

Aspergillus luchuensis Inui, in Jour. Col. Sci. Imp. Univ. Tokyo 15: 469,

PI. 22, fig. 1-8. 1901. See also Usami, in Centralb. Bakt.

etc., 2 Abt., 43, p. 250. 1915; The Aspergilli,

p. 171. PI. IV. 1926.

Colonies upon Czapek's solution agar spreading rapidly, producing
abundant conidiophores and conidial heads which give a purple-black color

230 A MANUAL OF THE ASPERGILLI

to the whole colony, reverse in pale yellow shades. Conidial heads globose,
up to 250 to 300m in diameter, splitting in age into short columns of spores.
Conidiophores up to 1500m by 10m, smooth, yellow toward the vesicle. Ves-
icles yellow, up to 40m in diameter, fertile over the entire surface. Ster-
igmata mostly in one series, 6m or a little larger by 3m ; branched sterigmata
occasionally appear. Conidia globose, 3.5 to 4m, roughened with spines
rather than bars of coloring substances.

Representatives of the species have been received from Japan (NRRL
No. 356; Thorn No. 4291.3), from West Africa, from Bermuda, and from
various points in the United States. While not as abundant as forms
with double sterigmata, isolates with this kind of head are not uncommon.

Inui also described A. perniciosus (Jour. Coll. Sci. Tokyo 15, p. 473,
T. XXI, figs. 9-12. 1901) with color data closer to A. wentii than A.
luchuensis. Its morphology, however, seems to belong here. It has not
been rediscovered and discussed adequately in relation to either group.
Culture No. 4707.757 (Thom), received from da Fonseca in Brazil, possibly
represents this species: colonies in center at least transiently greenish,
but without true green color; conidiophores not crowded, sinuous, apparently
smooth when observed with low magnifications but with traces of pitting
evident when examined with an oil-immersion objective, up to 1000m by
10 to 15m; primary sterigmata 10 to 16m by 3 to 4m, secondary up to 8m by
2 to 3m ; conidia 4 to 5m in diameter, with yellow markings in the form of
loops and bars.

A. luchuensis var. rubeolas Shih (Lingnan Sci. Jour. 15(3): 374. 1933)
differs from the species by becoming chocolate brown rather than black.
This would suggest careful comparison with A. japonicus Saito.

Aspergillus japonicus Saito, in Bot. Mag. (Tokyo) 20: 61, 5 figs. 1906.

Colonies upon Czapek's solution agar growing rapidly and spreading
evenly, characterized by its purple-brown heads and the presence of few
to many light brown sclerotia ; reverse colorless or nearly so. Conidiophores
500 to 1000m by 12 to 15m, figured as showing concretions on the surface,
with walls more or less brown. Vesicles globose, fertile over the whole
surface, with walls brown and marked by the bases of sterigmata. Ster-
igmata in one series, 7 to 9m by 5 to 6m, commonly falling away in mounts
from old cultures. Conidia globose, echinulate, 4 to 5m in diameter.
Sclerotia scattered throughout the colony, 650 to 1000m in diameter, white
to pale yellow in color and often overgrown by mycelium and conidiophores.

Type material has not been seen. The diagnosis is based upon two
strains contributed by Dr. A. F. Blakeslee (NRRL No. 358: Thom No.
4030.3, and NRRL No. 359: Thom No. 4030.5) which come fairly close to

ASPERGILLUS NIGER GROUP 231

Saito's description. A purple-brown strain (Thorn No. 5362.7) collected
by Manns in Honduras showed slightly darker colors and slightly different
measurements. A strain received from Raistrick as coming from Blochwitz
through the Centraalbureau in Baarn labeled A. alropurpareus Zimmermann
resembled the two Blakeslee cultures quite closely. It certainly did not
comply with Zimmermann 's description (see p. 226). A. luchuensis var.
rubeolis Shih is probably closely related if not identical with .4. japonicus
Saito.

Aspergillus violacco-fuscus Gasperini, in Atti. Soc. Toscana Sci. Nat. Pisa,

Mem. 8, fasc. 2, p. 326. 1887.

Colonies upon Czapek's solution agar, comparatively slow growing
(fig. 61 F), purplish-brown with a faint violet shade, passing to purple drab
in age, reverse colorless to dark purplish. Conidial heads purplish-brown,
globose, not crowded, 100 to 150m in diameter. Conidiophores mostly
less than 1 mm. in length but sometimes reaching 2 mm., 12 to 18m in di-
ameter. Vesicles globose, varying up to 60m in diameter. Sterigmata
generally in one series (fig. 62 D), 5 to 8m by 3m, occasionally in two series
with secondaries 2 to 4m long. Conidia elliptical, 3.5 to 5m by 5 to 6.5m, at
first hyaline, becoming violaceous, somewhat roughened.

By description this is a variant member of the great group of black
Aspergilli characterized by short sterigmata and elliptical conic|ia.

Gasperini's material has not been seen but three cultures have been
studied which are believed to differ from his species only in having some-
what smaller conidia. The first of these was received in 1914, from Puerto
Rico (NRRL No. 360: Thorn No. 3522.30), while the others were subse-
quently obtained from Jamaica and from Professor Raistrick in England.

In cultivating black Aspergilli an occasional strain produces heads at
first showing a single series of sterigmata; then, as the colony becomes
older, heads with both primary and secondary sterigmata dominate the
culture. Upon careful examination many strains show both large heads
with sterigmata in two series and, on shorter conidiophores mixed among
the larger ones, small heads with simple sterigmata only or with both
types mixed. Colonies of this kind probably accounted for .4. nanus
Montagne, .4. subfuscus Johan-Olsen (Sopp.) and are known to account for
A. pyri English.

Aspergillus nanus Montagne, in Syllog. Generum Specierumque Cryptogamarum,
p. 300, No. 112, Paris, 1856. Sacc. Syll. 4: 71. 1886. Species reported as a member
of the black group with a single series of sterigmata about 15m in length and spores
3m in diameter. This may have represented young fruiting structures of a typical
A. niger.

Aspergillus subfuscus Johan-Olsen, in Meddelelser fra Xaturh. forening i Kris-

232 A MANUAL OF THE ASPERGILLI

tiania, 1885. Described as showing smooth globose spores, 3.0 to 3.5m and a single
series of sterigmata in culture, up to 20m in length, but with some secondary sterig-
mata seen in the original material. The separation of this from A. niger appears
questionable.

Aspergillus pyri English n. n., in Doctoral Thesis, State College of Washington,
Pullman, Wash. pp. 76-78, 1940; cited in abstract. A form described as showing a
single series of sterigmata 14.4 to 19.2m by 3.6m and spinulose conidia 3.6 to 4.8m in
diameter. In personal correspondence of June 1943, English stated that as a result
of continued study of this strain and the finding of double sterigmata, he ques-
tioned the desirability of maintaining the species designation.

SYNOPSIS OF SPECIES PROPOSED BY MOSSERAY

Biourge, who was a discriminating collector, accumulated 63 strains of
"black" (or related) Aspergilli for each of which he could see sufficient
individuality to warrant preservation. Mosseray, working in Biourge's
laboratory, studied these strains, attempted to establish species lines among
them, and proposed new specific names for all forms which he believed
undescribed. With the whole 63 strains in parallel culture, all bearing the
names applied by Mosseray, Biourge and Simonart were inclined to with-
draw part of Mosseray's new species names, a position with which the
writers heartily agree. Nevertheless, to present one concept of the range
of variation confronted by the student of this group, Mosseray's synopsis2
has been translated with minor emendations, and is herewith presented.

Mosseray's Synopsis

A. Conidia 6 to 10m in diameter, rough; vesicles subglobose; primary sterigmata often
100m or more in length; colonies jet-black.

a. Sporulation more or less dense ; heads large ; reverse fumose or very dark olive

with mycelium more or less wrinkled.
Conidiophores 2, 4, or even 6 mm. long; sclerotia present in "natural" media

A. carbonarius (Bainier) Thorn and Currie

Conidiophores 1 to 2 mm. long; sclerotia not reported A. pulchellus (Speg.)

Thorn and Church, Syn. S. acini-uvae Caballero

b. Sporulation less dense; heads small, also with very small heads with single

2 Translated and emended from Mosseray, Raoul, Les Aspergillus de la section
"niger" Thorn and Church, in La Cellule XLIII: 271-273. 1934. All data are based
upon colonies grown in slanted test tubes using Biourge's formula of "neutral Raulin
agar" of the following composition:

Water (distilled) 1000 cc . (NH4) 2HP04 0.400 g.

Sucrose 50 g. (NH4)2S04 0.200 g.

Tartaric acid 0.40 g. FeS04 cryst 0.050 g.

MgCC-3 0.250 g. ZnS04 0.050 g.

NH4N03 2.500 g. Agar 20.0 g.

K2C03 0.400 g.

Sterilized at 120°C. for 20 minutes.

ASPERGILLUS NIGER GROUP 233

sterigmata and on very short conidiophores among them; mycelium gray-
yellow; reverse dark reddish-brown and mycelium much wrinkled

A. pseudo-carbonarius (Bainier nn.) Mosseray

B. Conidia 2.5-4, even to 5m in diameter, mostly more or less rough and more or less
colored but some smooth or nearly so ; vesicles normally globose ; primary sterig-
mata mostly 20m long or longer, sometimes up to 100m.
a. Colonies deep purple-brown or clear purple-brown.
I. Conidiophores short (up to 2 mm. on an average).

a. Wrinkles in reverse shallow, transverse (in tubes).

Heads small ; conidiophores short (up to 1 mm.) 8 to 15m in diameter,
primary sterigmata 5 to 20m; reverse cream colored

A. microcephalus Mosseray
Heads "medium"; conidiophores up to 2.5 mm.; primary sterigmata
up to 45m; reverse dark reddish-brown, often spotted and becom-
ing very dark A. phoenicis (Corda) Thorn

Syn. A. longobasidia (Bainier n.n.) Mosseray

Syn. A. bainieri Mosseray 1934

/3. Wrinkles in reverse of mycelium fairly numerous, occasionally

anastomosing.

Appearance "mealy", deep purple-brown; conidiophores very short

up to 0.5 mm. and 7 to 13m in diameter; primary sterigmata 4 to

12m; reverse deep olive-brown A. pseudo-citricus Mosseray

Appearance sub-granular; brown-purple, at times clear brown to
umber; conidiophores up to 1.5 mm. by 7 to 20m; conidia smooth or
nearly so; growth rapid. Reverse slightly colored:
Reverse cream to pale brown; primary sterigmata 8 to 20m

A . fuliginosus Peck

Syn. S.fuliginosa Bainier

Syn. A. praecox Mosseray 1934a

Reverse very deep brown; primary sterigmata up to 50m; sclerotia

occasional A. sclerotifer Mosseray

Reverse uncolored to citron yellow; primary sterigmata 8 to 20m

A. citrino-niger Mosseray
7. Wrinkles sharp and numerous (reticulated). Reverse becoming
rapidly dark, almost black.

Aspect mealy; color purple-brown; conidiophores up to 1 mm;
drops not colored; primary sterigmata 12 to 30m; conidia smooth

or nearly so A . densus Mosseray

Aspect mealy to granulate, deep purple-brown; drops numerous,
bronze; primary sterigmata 12 to 30m. • -A. rutilans Mosseray
Aspect subgranular, brown-purple; drops numerous, black, some
of them very large; primary sterigmata 12 to 30m

A. guttifer Mosseray
Reverse not so dark, sometimes olive or colorless.
Reverse deep olive, often spotted; primary sterigmata 15 to 30m

A. Buntingii Mosseray
Reverse not colored; commonly showing areas sterile or free
from spores; mycelium slightly yellow at first; primary sterig-
mata 20 to 50m A. variegatus Mosseray

234 A MANUAL OF THE ASPERGILLI

II. Conidiophores up to 3 mm., rarely 4 mm. long.
a.. Sporulation normal.

Reverse uncolored or slightly olive; drops few or none; primary
sterigmata 20 to 40m; mycelium often yellow at first

A. niger Van Tiegham
Reverse orange-brown or purple-brown; drops large, black; pri-
mary sterigmata 8 to 25m; mycelium colorless or rarely yellow

A. Biourgei Mosseray

Reverse pale reddish-brown; drops very few; heads very small;

primary sterigmata 15 to 30m; mycelium colorless or sometimes

rose; sporulation very slow A. Churchii Mosseray

Reverse golden yellow; drops none; primary sterigmata 20 to 40m;
mycelium reddish-yellow at first

A. luteo-niger (Lutz) Thorn and Church
Reverse dark olive-brown; conidiophores up to 1 to 2 mm.; primary
sterigmata very variable; vesicles subglobose

A. anomalus Mosseray

Reverse cream to brown, spotted slightly, wrinkled with a dark

band in center, sporulation scattered, pale, conidiophores up to

4 mm. long A. tubingensis (Schober) Mosseray

/3. Sporulation abnormal.

Sporulation absent in patches and in the margin, with a granular
appearance, and proliferation of mycelium throughout the co-

nidial area A. granulatus Mosseray

Sporulation massed at the thin end of the agar, less dense toward

the bottom of the tube; mycelium wooly.
Mycelium wooly, gray-white or yellowish-gray, carrying on the
thin areas numerous simple heads ("fumigatiformes") and some
normal heads. Reverse cream, slightly wrinkled

A. velutinus Mosseray
Mycelium less wooly, white; conidiophores 1 to 3 mm. long, very
abundant at the thin end of the agar, the remainder sterile. Re-
verse much wrinkled, cream. .... A . ficuum (Reich.) Hennings
III. Conidiophores tall, up to 1 cm.; sporulation scanty, mostly toward the
thin end of the agar.

Heads large, splitting into divergent columns; conidiophores up to
1 cm. by 30m; primary sterigmata up to 100m; reverse cream or slightly

rose A . elalior Mosseray.

Heads small, mycelium gray-rose; conidiophores rarely up to 1 cm.,
more often 2 to 6 mm.; very numerous little "fumigatiform" heads, on
short conidiophores at the surface; reverse clear rose or salmon

A. pseudo-elatior Mosseray

IV. Conidiophores very irregular from less than 1 mm. to 3 mm. with much

larger heads; reverse clear reddish-brown; mycelium yellow at first

then dark brownish-red; sporulation delayed. A. pseudo-niger Mosseray

b. Colonies not purple-brown but clear brown; morphology of A. niger; conidia

mostly smooth or nearly so.

I. Colonies clear brown or sepia; conidiophores up to 1 mm. ; reverse olive or
lighter; with reticulated wrinkles; vesicles 20 to 50m; primary sterig-
mata 12 to 50m; conidia smooth A. olivaceo-fuscus Mosseray

ASPERGILLUS NIGER GROUP 235

II. Colonies umber; conidiophores 1 to 3 mm.; reverse reticulated, dark

reddish-brown; conidia smooth or nearly so A. Schiemanni Thorn

Syn. A. fuscus Schiemann
III. Colonies reddish salmon; conidiophores 1, 2, or even 3 mm.; reverse
slightly wrinkled, uncolored, or pale cream; conidia smooth

A. cinnamomeus Schiemann

C. Conidia 3 to 5m, globose, smooth, slightly colored; vesicles globose; conidiophores

up to 1 mm., slender ; primary sterigmata 12 to 20m; colonies appearing somewhat
granular, deep brown; reverse olive passing to bronze; mycelium sulphur yellow
at first A . citricus (Wehmer) Mosseray

D. Conidia 3 to 5m, globose or elliptical, smooth or slightly rough ; colonies violaceous.

a. Sterigmata in two series; vesicles globose; colonies purplish-violet or mauve;

reverse violet-brown: conidia 3 to 5m, globose, smooth, uncolored

A. awamori Usami

b. Sterigmata in one series, short; vesicles subglobose; colonies in violaceous

shades to mauve.

Colonies mauve ("violet livide"), reverse uncolored, wrinkled; conidia
globose or obovate, smooth 3 to 5m in long axis; sporulation slow; narrowly
growing A . malvaceus Mosseray

Colonies violaceous or dark violet slate; reverse dark yellow or orange,
slightly wrinkled; conidia globose and rough, 3 to 4.5m; sporulation very
rapid, and colonies broadly spreading, with sclerotia common on rice or
other "natural" substrata, rare upon sugar media A.japonicus Saito

Colonies violet-brown; reverse purplish-brown, wrinkled; conidia globose,
rough, 3 to 5m in long axis; sporulation slow and more or less incompletely
covering the surface; dwarf heads abundant. .A. atro-violaceus Mosseray

Colonies dark brown or carob brown, with a mealy or granular appearance,
reverse dark brown, or olive at the margins, wrinkled; conidia smooth,
globose, 2.5 to 4m; vesicles globose or pyriform; primary sterigmata 3.5 to
7.5m in diameter -4.. atro-fuscus Mosseray

If the material available for study were limited to Biourge's 63 cultures,
identification to strain, variety, or species might be possible. When Mos-
seray subsequently returned to Brussels as Mycologist at the Jardin
Botanique de l'Etat and was confronted by hundreds of other strains
largely from the Belgian Congo, many of which presented further varia-
tion, he began to see the impossibility of describing them all in terms which
would permit subsequent identification. Biourge at that point (personal
conference) withdrew his support from many of the diagnostic features
accepted in Mosseray's memoire and agreed to the proposal that specific
names in black Aspergilli could only be serviceable as bringing together
aggregates of strains showing common and fairly dependable morphological
characters. This was the attitude held by Thorn and Church in The
Aspergilli (1926) based upon the examination of many hundreds of black
Aspergilli, and it remains the position of the writers at this time.

Since variation is characteristic of the whole genus — not one group

236 A MANUAL OF THE ASPERGILLI

alone, the subject is covered in the chapters on Morphology and on Varia-
tion.

Coloration

Color has been emphasized in most of our attempts to separate these
strains in culture. Linossier (1891) extracted from his strain of A. niger
a coloring substance soluble in hot water which he called aspergilline.
This substance is readily demonstrated. Blochwitz (1929, p. 219) reports
this material to be soluble in "NH3 and alkalies," and we have extracted it
with hot water. Microscopic comparison of conidiophores, heads, and
separate conidia from numerous strains leads to the conclusion that the
same substance may give a brown color to the upper one-third of the
conidiophore, to the vesicle and its contents, to the walls of the sterigmata,
and in A. carbonarius so fill the whole vesicle and sterigmatic area with a
brittle mass as to justify Bainier in calling the head carbonaceous. Conidia
in a few strains have appeared to possess smooth, uniformly brown cell
walls. As a group, however, the black aspergilli have globose conidia with
a firm, uncolored or slightly colored inner wall, a very thin outer wall, and
between the two the coloring substance, presumably aspergilline, deposited
as granules, warts, or bars running lengthwise of the cells and presenting
a pattern characteristic of the whole group. When pale-colored variants
(mutants?) such as A. cinnamomeus (p. 223), or darker ones such as A.
schiemanni (p. 224), or the variously brown to deep carbon black ones
like A. carbonarius are compared, the relative quantities of coloring matter
seem to account for the progressive darkening of the head colors as de-
scribed in the series.

In the "reverse", or underside, of the mycelium and in the culture
substratum, a range of shades from colorless to yellows to reddish-brown,
and even very dark shades, is reported for particular species growing upon
specified substrata. The chemical reactions back of these colors are not
known for the black aspergilli. Blochwitz has reported extractions of
color from various species of Aspergillus and other molds, and observed
that variations in these colors were readily obtained with reagents. Some
of the extracted materials were reported to act as indicators. It is, there-
fore, doubtful whether the succession of colors present in such a related
series of organisms represent substances differing .fundamentally one from
another, or merely successive steps in the transformation and reactions of
the same general type of product.

Occurrence and Economic Importance

The black aspergilli are probably more common than any other repre-
sentatives of the genus. They are world-wide in distribution and occur

ASPERGILLUS NIGER GROUP 237

in and upon the greatest variety of substrata, including grains, forage
products, spoiled fruits and vegetables, exposed cotton textiles and fabrics,
leather, dairy products and other protein-rich substrata, and decaying
vegetation in the field. They are abundant in all soils examined; and from
studies which have been made by the authors and other investigators, it
would appear that they are particularly abundant in soils from tropical and
sub-tropical areas.

With the possible exception of the A. flavus-oryzae group, which is of
great economic importance in the Orient, the black aspergilli are un-
doubtedly more widely used in industry than any other group of molds.
Since the present volume is primarily a manual designed to assist the
worker in the study, diagnosis, and maintenance of the aspergilli that come
into his hands, no attempt will be made to discuss the various fermentations
and other biochemical activities of the black aspergilli. However, these
fermentations are of great importance and will be briefly noted. For the
reader who is interested in these fermentations, or perchance, is actually
conducting them, a fairly complete list of references is presented for each
in the "Topical Bibliography" (Chapter XXII). In each case it has been
our aim to present sufficient references to provide the reader with a reason-
ably comprehensive guide to the literature of the field.

Gallic Acid: Raulin and his coworkers in Paris during the early 1860's
identified the organism active in the production of gallic acid by the fermenta-
tion of gallnuts and other tannin-bearing substances. The species was
first discussed as Ascophora nigrans. Van Tieghem in 1867, named and
described the organism correctly as A. niger. Recurrent investigations
have been conducted on this fermentation from that period to the present
time (see Topical Bibliography, p. 294).

Citric Acid: In 1917 Currie published his fundamental studies on the
formation of citric acid by strains of A. niger and thereby established the
basis for one of the most important of all industrial mold fermentations.
Subsequent to this, investigators in the United States and abroad have
made many additional and important contributions. While no attempt
will be made to cite all of these, the works of Bernhauer, Wehmer, Doelger
and Prescott, and the U. S. Department of Agriculture group, including
Wells, May, Moyer, Herrick, and Ward, are considered to be outstanding.
A selected list of references to the citric acid fermentation is presented
on pages 290-293.

Fumaric Acid: Certain strains of the A. niger group produce appreciable
amounts of fumaric acid, and it was to one of these forms that Wehmer in
1918 applied the n&me A. fumaricus. At the present time, however, fumaric
acid is produced in industry by fermentation with species of Rhizopus
rather than strains of the black aspergilli (see p. 293).

238 A MANUAL OF THE ASPERGILLI

Gluconic Acid: Selected strains of A. niger are used industrially for the
production of gluconic acid. This process, like the citric fermentation,
has been investigated by many workers. Outstanding contributions have
been made by Molliard, Bernhauer, and the U. S. Department of Agri-
culture group, including Herrick, May, Wells, Moyer, Gastrock, Porges,
and others (see pp. 295-297).

Oxalic Acid: Under certain conditions some strains of A. niger produce
appreciable quantities of oxalic acid. While it is usually avoided rather
than encouraged, this fermentation has been investigated by Wehmer (1891
and 1892), Raistrick and Clark (1919), Jacquot (1938), and others (see
pp. 298-299).

The production of these various acids in quantities sufficient to have
economic importance represents to an appreciable degree specific strain
characteristics, and the greatest possible care must be exercised in main-
taining these strains in a state of high productivity. It has been found,
however, that in certain cases the same strains can be made to produce
substantial yields of two or more of these acids by varying the composition
of the nutrient solution and certain environmental factors. The reader is
referred to the extensive literature on this subject.

Enzymes: While they are not commonly cultivated for the production
of enzymes as such, as are members of the A. flavus-oryzae group, the black
aspergilli produce a number of enzymes in appreciable quantities. Be-
ginning with Fernbach in Germany (1890) and Bourquolet (1893) in
France, various authors have devoted considerable study to their formation.
A number of papers relative to enzyme production by members of the group
are cited in the "Topical Bibliography" under the subtitle "Enzymes of
Aspergillus niger" (pp. 294-295).

Fat Production: The mycelium of A. niger contains appreciable fatty
materials (Pontillon, 1932; Bernhauer and Patzelt, 1935; Schmidt, 1935;
and others). The waste mycelium from the citric acid fermentation is
reported to provide a satisfactory source of sterols for irradiation in the
production of Vitamin D.

Soil Testing: A. niger has been successfully employed as an assay
organism for determining mineral deficiency of soils, particularly de-
ficiencies in phosphorus and potassium. Papers on the so-called A. niger
method of soil testing were first published by Kiessling and Schmidt and
by Schlots, Smith, and Brown in the same year (1932), to be followed by
more elaborate studies by Stock (1933), Niklas and associates (1933), and
others. Additional references to this method are presented in the "Topical
Bibliography" (pp. 313-314). The strain employed by Niklas is main-
tained in the culture collection of the Northern Regional Research Lab-
oratory as No. 323.

ASPERGILLUS NIGER GROUP 239

Mildew: Strains of A. niger represent a common cause of mildew on ex-
posed wood surfaces and cotton fabrics. Partansky and McPherson
(1940) used a strain of this species successfully for testing the mold-
resistant properties of oil paints. Aspergillus niger is commonly included
in the mixtures of miscellaneous molds used for testing the effectiveness
of mildew- and rot-proofing agents when impregnated in textiles and fabrics.
Where pure cultures are employed, species of Chaetomium and Metarrhizium
are generally used.

Mold Physiology

Members of the Aspergillus niger group have been used extensively in
investigations on mold physiology, probably more than any other form.
As early as 1909 Latham studied nitrogen assimilation by A. niger (S.
nigra) to be followed in 1911 by Dox studying phosphorus assimilation by
the same species. Beginning in 1918 and continuing up to the present
time, Steinberg has published a succession of papers on the physiology of A .
niger, with special reference to the role of heavy metals in its nutrition.
A single strain of A. niger which is carried in the NRRL collection as No. 334
(Thorn No. 4247) has been used throughout these investigations. Studies
of a somewhat similar character have been conducted by Bortels (1927),
Levy (1932), Gollmick (1936), and others. Citation of these papers, to-
gether with many additional references are presented in the Topical
Bibliography under the subtitle "Physiology". An attempt has been
made to present sufficient references to serve as a point of entrance to the
literature of the field.

Pathogenesis

Members of the Aspergillus niger group are commonly isolated from
the external ear of man, this being the source of the classic Sterigmatocystis
antacustica of Cramer. Other species reported to have been isolated from
cases of otomycosis include A. niger van Tieghem, A. phoenicis (Cda.) Thorn
and Church (with long primary sterigmata), A. giganteus (Mattlet) Dodge,
and A. Macfiei Dodge. A. Macfiei showed no signs of pathogenicity
when tested on experimental animals. There is no morphology to dis-
tinguish either of the latter two forms from the ubiquitous saprophytic
types, and it appears probable that many strains can become established
in the auditory canal under certain favorable and probably temporary
conditions.

Once entrenched in the flesh about the auditory canal, the mycelium has
been found to be very persistent. One case is known in which occasional
abscesses have occurred over a period of 25 years during which desultory
treatment has quieted the inflammation but has not destroyed the parasite.

240 A MANUAL OF THE ASPERGILLI

In the same way, extensive development of these forms as points of
infection in the lungs may give rise to conditions diagnosed as pseudo-
tuberculosis which may persist for long periods without resulting either in
death or complete recovery of the patient. Spores of A. niger are air
borne and hence are commonly drawn into the respiratory tract. They are
occasionally reported as causative agents in allergic reactions.

Single Strain Cultures

The great importance of some of the black aspergilli as fermentative
agents makes punctilious preservation of the actual strain employed the
only means either of insuring a process against breakdown or of intro-
ducing it in a new locality. Even when such a culture is maintained with
the utmost care, natural variation sometimes occurs. Induced variation,
such as that described by Steinberg and Thom in a series of papers (1939-40)
presents a further hazard in processes where cultures are grown at extreme
H-ion concentrations, or in the presence of chemical or other substances
which may actually be toxic at the concentrations employed. In a process
in one laboratory a strain appeared that presented a colony aspect in which
only a few typical black heads developed in a background of dwarfed and
fractional conidial fruiting structures. In another case, a mutant appeared
which differed so radically from A. niger in its secondary sterigmata and
spore chains that the Yuills described it as a new genus and species, Clado-
sarum olivaceum (Trans. Brit. Myc. Soc. 22: 194-200, Pis. 11-13. 1938).

The user of one of these Aspergilli, then, needs to know his organism in
cultural aspect, in microscopic details, and in essential reactions in standard-
ized and reproducible substrata. Furthermore, he must be able to main-
tain it free from contamination with other species, and likewise free from
such variation, either natural or induced, as will interfere with consistent
and controlled results.

Chapter XVIII
THE ASPERGILLUS WENTII GROUP

Outstanding Characters

Conidial heads large, typically globose, often splitting irregularly in
age — varying in color from dull yellowish to ecru-olive, and from light
to dark brown depending upon the species and strain.

Conidiophores smooth-walled or nearly so, but often appearing finely
roughened when examined in dry mounts.

Vesicles globose, fertile over the entire surface. Sterigmata in two
series.

Conidia commonly elliptical, smooth or somewhat roughened depending
upon the species.

Sclerotia present or lacking, dark brown to black, characteristically
white-tipped when young.

The Aspergillus wentii group as presented here is recognized as somewhat
artificial, in comparison with such strictly natural groupings as the A.
glaucus, A. nidulans, and A. clavatus groups. The degree of relationship
between the species included is open to question. Yet all of the forms
possess certain characteristics in common: (1) conidiophores are smooth-
walled, or nearly so; (2) conidial heads are large and strictly globose, at
least when young; and (3) all appear to occupy taxonomic positions some-
what intermediate between the Aspergillus niger group on the one hand
and the Aspergillus flavus or Aspergillus ochraceus groups on the other.

Group Key

I. Conidia smooth-walled.

A. Sclerotia lacking; vegetative mycelium and young conidiophores reddish in

color.

1. Conidial heads light brown, near wood brown (Ridgway PI. XL)

A. panamensis Raper and Thorn

B. Sclerotia present, dark brown to black, vegetative mycelium colorless, and

young conidiophores colorless or pale yellow.

1. Conidial heads dull yellow to ochraceous; sclerotia globose or nearly so

A. alliaceus Thorn and Church

2. Conidial heads in yellow-green shades near ecru-olive (Ridgway PL

XXX) A. avenaceus G. Smith

II. Conidia more or less echinulate.

A. Sclerotia present or lacking, depending upon the strain and the substratum;
conidial areas in orange-brown to brown colors.

1. Conidiophores colorless; colonies often conspicuously floccose

A . wentii Wehmer

2. Conidiophores brown (See A. niger) A. hennebergi Blochwitz

241

242 A MANUAL OF THE ASPERGILLI

Aspergillus panamensis Raper and Thorn, in Mycologia 36: 568-572,

fig. 5. 1944.

Colonies on Czapek's solution agar at room temperature very thin,
consisting of a sparse and transparent growth of vegetative hyphae, almost
wholly submerged, bearing widely scattered, erect conidial structures
with radiate heads, light brown in color. Colonies upon malt extract agar
at room temperature growing well and fruiting luxuriantly, reddish brown
in color, consisting of a dense basal mycelium, predominantly red, from which
develop massed condial structures in broken or continuous concentric
zones (fig. 65 A), many conidiophores abortive and sterile, fertile co-
nidiophores bearing globose to radiate heads, light brown in color, near wood
brown (Ridgway, PI. XL) ; these, together with red-colored sterile structures
and aerial hyphae, give the colony its characteristic appearance and color;
in age, colonies tending to develop a loose floccose overgrowth, more or
less obscuring the abundant conidial heads; reverse dull brown; odor none.

Conidial structures arising directly from the substratum, scattered or
abundant, depending upon the culture medium employed (fig. 65 B).
Heads typically globose, in age characterized by loosely radiating chains
of conidia, less commonly by few to several roughly columnar masses variable
in size, commonly ranging from 250 to 450m in diameter, occasionally up
to 500m, varying in color from avellaneous to wood brown (Ridgway, PL XL)
to Saccardo's umber (Ridgway, PI. XXIX). Conidiophores straight,
mostly 600 to 900m in length by 9 to 12m in diameter, occasionally larger,
with walls smooth, comparatively heavy, ranging from 3 to 3.5m thick in
the basal area to 1.5 to 2m in the terminal area, approximately uniform in
diameter throughout except for a limited reduction immediately beneath
the vesicle. Vesicle colorless, comparatively thin-walled, globose or
slightly elongate, mostly 25 to 30m in diameter, fertile over the entire area
(fig. 65 C). Sterigmata in two series, closely packed, primaries 5.5. to
6.5m by 2.4 to 2.8m, secondaries 5 to 6m by 1.5 to 2m. Conidia light yellowish-
brown in mass, globose to subglobose, smooth-wal'ed, mostly 2.2 to 2.6m
in diameter, occasionally 2.8m-

Type culture NRRL No. 1785 was isolated in January 1942, from
Panama soil collected by Mr. John T. Bonner. A second culture, NRRL
No. 1786, differs from the above strain in minor details but clearly belongs
with it. This was isolated from a second sample of Panama soil collected
by Bonner.

The species is considered to represent a form somewhat intermediate
between the Aspergillus nigcr group and A. wentii. Superficially, at
least, there is evidence of relationship with Aspergillus niger mut. cin-
namomeus (syn. Aspergillus cinnamomeus Schiemann) and Aspergillus
niger mut. Schiemanni (syn. A. fuscus Schiemann). It bears a certain

Fig. 65. A-C, Aspergillus panamensis NIHIL No. 1785: A, Colonies upon malt
extract agar, 10 days; B, Conidial heads, showing tendency to split into loose diver-
gent columns, X 9; C, Conidial heads showing globose vesicles and sterigmata in
two series, X 500. D-F, Aspergillus avenaceus NRRL No. 517: D, Single colony
on Czapek's solution agar snowing numerous large sclerotia; E, Portion of above
colony showing greater detail of heads and sclerotia, X 6; F, conidial heads showing
large globose vesicles, X 160.

243

244 A MANUAL OF THE ASPERGILLI

resemblance to these forms in the comparatively light color of its conidial
heads and in the smallness and general character of its conidia, but differs
from these forms in three very striking particulars. (1) It character-
istically develops an extensive red-colored aerial mycelium upon media
such as malt-extract agar where it attains its maximum growth; (2) it grows
very sparsely upon Czapek's solution agar, upon which the above noted
forms grow luxuriantly; and (3) it possesses very small primary sterigmata,
measuring 5.5 to 6.5m by 2.4 to 2.8m in contrast to 13 to 15m by 3 to 5m for
mut. cinnamomeus and 15 to 40m by 4.6m for mut. Schiemanni. Whether
or not the species actually represents a naturally occuring mutation from
A. niger can only be guessed. The smallness of its conidia and primary
sterigmata would hardly support this hypothesis. In cases where mu-
tations have been obtained from known cultures, the dimension of specific
structures in such mutations generally agree very closely with those of the
same structures in the parent strain; and black Aspergilli with the dimen-
sions of A. panamensis are rarely, if ever, encountered in nature. The
possibility of this representing a mutation is not excluded, but until ad-
ditional evidence supporting such origin is forthcoming, the writers feel
warranted in maintaining as a distinct species this unique form which
obviously is able to maintain itself in the soils of Panama.

The correct taxonomic position of this species remains in doubt. It is
included with Aspergillus wentii, although we realize that this placement
is not entirely satisfactory. As continued isolations are made from tropical
soils and other sources it is our hope that additional forms may be found,
which will furnish evidence of a more exact relationship.

The very sparse development of A. panamensis upon Czapek's solution
agar, containing sucrose, results from an invertase deficiency; when dextrose
is substituted as a carbon source the fungus grows luxuriantly and fruits
abundantly.

Aspergillus alliaceus Thom and Church, in The Aspergilli, p. 163. 1926.
Discussed without name as Thom No. 4660 by Walker and Lindegren,
in Jour. Agr. Res. 29: 507-514. 1924; and by Walker, Lindegren, and
Bachmann, in Jour. Agr. Res. 30: 175-187. 1925.

Colonies on Czapek's solution agar rapidly and broadly spreading, with
loosely floccose aerial sterile mycelium, bearing scattered ochraceous
heads among abundant dark to almost black sclerotia (PI. VI D and fig. 66
A) in some strains, predominantly floccose with limited conidial heads and
few sclerotia in others (fig. 66 B); reverse uncolored. Conidial heads dull
yellow to ochraceous, strictly globose when young and remaining radiate
or splitting irregularly in age (fig. 66 D), up to 300m in diameter, often more
abundant in cultures after many transfers. Conidiophores up to 150m

THE ASPERGILLUS WENTII GROUP

245

long by 15m, with walls up to 1.5m in thickness, appearing smooth in liquid
mounts, but showing rudimentary markings or pits when examined dry.
Vesicles globose to subglobose, up to 40 to 50m in diameter, with walls
about 2m in thickness, and showing pores where sterigmata are attached.
Sterigmata in two series, primary 7 to 9m or even 12m by 3 to 4m, secondary

A

B

Fig. 66. Aspergillus alliaceus. A, Heavy sclerotium-producing strain, NRRL

(strain NRRL No. 318) showing tendency to split into divergent columns, X 18

7 to 8m by 2m- Conidia elliptical to globose, yellowish, 2.5 or 3m in diameter.
Perithecia not found. Sclerotia often very abundant (fig. 66 C) at first
white but quickly becoming black or nearly so, in many strains dominating
the character of the colony.

Strains compared include NRRL Xo. 315 (Thorn No. 4656) from a dead

246 A MANUAL OF THE ASPERGILLI

blister beetle (Macrobasis albida), NRRL Xo. 316 (Thorn No. 4660)
isolated from onion bulbs, and others with characteristic heads and scle-
rotia have been obtained from garlic bulbs, from cactus plants, and from
soils, particularly from the general region of Texas, Arizona, and Mexico.
Strains have also been isolated from soils collected near Calcutta.

The large, globose, pale yellow to ochraceous heads of the species strongly
suggest relationship to the A. oehraceus group. The smooth-walled co-
nidiophores and black sclerotia, however, more closely ally it with Asper-
gillus wentii and the other species grouped with it. It may, however
represent a form somewhat transitional between the great groups repre-
sented by A. niger on the one hand and A. oehraceus on the other.

Aspergillus avenaceus Geo. Smith, in Brit. Mycol. Soc. Trans. 25: 24-27,

PL 1, figs. 1-3. 1943.

Colonies on Czapek's solution agar (with sucrose) at room temperature
spreading rapidly, more or less conspicuously zonate (fig. 65 D), slightly
fioccose, white at first, then dull yellow to ecru-olive (Ridgway, PL XXX)
as shown in PL VI E, with, at times, a greenish tinge without becoming
truly green; reverse pale dirty pink. Conidial heads large, globose 400
to 600m in diameter, or up to 1,000m, splitting into columnar masses of
conidial chains. Conidiophores up to 5 mm. long, 18 to 30m in diameter,
with walls 2.5 to 4m thick, smooth in fluid mounts; but appearing finely
roughened when examined dry. Vesicles globose or slightly flattened,
thick-walled, up to 185m in long axis, sterigmata in two series, primary 22
to 50m by 6m, secondary 11 to 13m by 4m (fig. 65 F). Conidia ellipsoid,
smooth, 4 to 6 or 6.5m by 3.2 to 4m- Sclerotia dark grayish-brown to
black (fig. 65 E), elongate, irregularly flask-shaped, sometimes with the
"neck" forked, apical portion white to gray during development, 2 to 3 mm.
in long axis, scattered in concentric zones after 7 to 10 days.

On Czapek agar with glucose, sclerotia are more abundant and larger.
On wort, or potato agar, conidial heads are abundantly produced but
sclerotia are delayed for several weeks and are few in number.

Species characterization adapted from George Smith's description.

This very distinctive species (NRRL 517: Thom 5725) is represented by
a single isolation from seed peas made in 1938 by Dr. G. E. Turfitt of the
London School of Hygiene and Tropical Medicine, University of London.

Aspergillus wentii Wehmer, in Centralbl. f. Bakt. etc., 2 Abt., 2, p. 150.
1896. See also The Aspergilli, Thom and Church, p. 183. 1926.

Colonies on Czapek's solution agar, rapidly growing and broadly spread-
ing, fioccose with white or yellowish aerial hyphae which in some strains
pile up in the plate (PL VI F, and fig. 67 A) or fill the test tubes for several

THE ASPERGILLUS WENTI1 GROUP 247

centimeters (fig. 67 C), but remain inconspicuous in other strains (fig. 67 B) ;
with developing heads at first white through yellow shades to olive-brown,
medal bronze or snuff brown (Ridgway, Pis. IV, XVI, XXX, column 19,
and XXIX, column 15 K.), or, according to Wehmer, coffee brown to
chocolate brown ; reverse becoming reddish-brown in old cultures. Conidial
heads large, globose, generally remaining radiate in age (fig. 67 D), ranging
up to 500/x in diameter, changing from yellow shades to brown. Conidio-
phores up to several millimeters in height by 10 to 25m in diameter, with
walls colorless up to 4m in thickness, studded with droplets in growing
colonies and often appearing slightly roughened when examined dry,
but uniformly smooth in fluid mounts. Vesicles globose or nearly so
(fig. 67 E), varying up to 80m in diameter, fertile over the entire surface.
Sterigmata usually in two series, primaries 10 to 20m by 3 to 5m, occasionally
much larger, secondaries 6 to 8m by 3/jl. Conidia borne in long chains,
more or less elliptical, ranging from 3.5 to 6m in long axis, but mostly 4
to 5m, double wall clearly evident, ranging from almost smooth to marked
by ridges sometimes suggestive of A. niger, again more closely resembling
the A. flavus series. No perithecia reported. Sclerotia often encountered
(fig. 67 F), dark brown to black, ovate with long axis vertical.

Culture description based upon strain NRRL No. 375 (Thorn No. 116)
obtained in 1909 from the Centraalbureau as Wehmer's original organism,
as well as numerous isolations from soils and other materials collected by
the authors in the United States, and other strains contributed by in-
vestigators from all over the world.

Numerous strains with the general aspect of Wehmer's species have been
seen from Java, China, South America, Japan, the Straits Settlements,
British Guiana, and Brazil. In our experience it has been isolated from
cottonseed cake, from olives, from soil, and from numerous other sources.
It is to be regarded as very widely distributed and to be common on many
types of decaying vegetable products.

The variations in colony aspect in different strains run from an extreme
of mycelial growth filling the test tube that is characteristic of cultures
such as the Wehmer organism, to colonies forming a crowded surface
growth of conidiophores only and distinguishable from .4. tamarii only by
a lack of greenish color in the early fruiting period and in the characteristic
smooth conidiophores and finely roughened conidia.

Aspergillus archaeoflavus Blochwitz (Ann. Mycol. 31(1/2) : 73-83. 1933) represents
a non-floccose form which is hardly separable from A. wentii. In our examination
of the type strain (XRRL No. 382: Thorn Xo. 5346), received in 1933 from Baarn,
measurements were somewhat less than those cited by the author. The absence of
conidial markings, to which Blochwitz called attention, would not bar it from A.
wentii since in some strains conidia are almost entirely smooth, in others finely rough-
ened, while in still others they are conspicuously echinulate.

248

A MANUAL OF THE ASPERGILLI

-

A

B

Fig. 67. Aspergillus wentii. A and B, colonies of strains NRRL No. 375 and No
385, respectively, upon Czapek's solution agar, 10 days. C, Tube cultures of four
representative strains. D, Conidial heads from old culture, X 18 E Single head
showing smooth conidiophore, globose vesicle, and sterigmata in two series, X 500.
t , Detail ! of colony margin in strain NRRL No. 379 showing sclerotia and abundant
conidial heads, X 9.

Plate VII

.4 (upper left), Aspergillus terricola var. americana Marchal, NRRL No. 424. B (upper right), Aspergillus
tamarii Kita, NRRL No. 427. C (center left), Aspergillus oryzae (Ahb.) Cohn, NRRL No. 458. D (center
right), Aspergillus flatus Link, NRRL No. 1957. E (lower left), Aspergillus quercinus (Bain.) Thorn and
Church, NRRL No. 394. F (lower right), Aspergillus ochraceus Wilhelm, NRRL No. 39s. All cultures
growing upon Czapek's solution agar. (Color photographs by Haines, Northern Regional Research Labora-
tory. Reproduced through co-operation of Chas. Pfizer & Co., Inc.)

THE ASPERGILLUS WENTII GROUP 249

Aspergillus wetitii var. minimus Xakazawa, Takeda, Okada, and Si mo (Jour.
Agr. Chem. Soc. Japan 10(2) : 176-177. 1934) shows measurements varying somewhat
from those of the species and from those given by Blochwitz, but it is not sufficiently
marked to warrant separation.

Sterigmatocystis aerect Bainier (Bull. Soc. Bot. France 27: 28. 1881) may have been
a member of this series but was not sufficiently described to permit positive
identification.

Aspergillus hennebergi Blochwitz, in Ann. Mycol. 33: 23S-239. 1935. The species
is described as having the aspect and colors of a non-floccose A. wentii or an A. tamarii
with red sclerotia but with conidiophores browned as in the partially browned con-
idiophores of the .4. niger group.

An albino "variant" of A. wentii was isolated by Mosseray (Ann. Soc. Sci. Brux.
54: 161-189. 1934) from a normal culture of this species, and was found to retain its
distinctive characters through repeated transfers in laboratory culture. No name
was given to this mutant.

Occurrence and Economic Importance

Aspergillus wentii is a cosmopolitan species that is fairly common in
soils, upon moist grains and other vegetable matter undergoing slow
decomposition, and may be isolated less frequently from a wide variety
of other materials collected from nature. It is apparently world-wide
in distribution. In the Orient, it is often included with Aspergillus tamarii,
A. flavus, and A. oryzae, all under the latter name as a rule, in the "Koji"
preparations used in the manufacture of various soy products. Likewise,
it has been investigated with these same species in connection with the
production of various mold enzymes. At the same time, it has been
included with the black Aspergilli in studies on the production of organic
acids by molds. Recently Karow (1942) has reported one strain of this
species to give substantial yields of citric acid in submerged culture.
Yabuta (1912) reports Koji acid production by .4. wentii. On the whole,
strains of the species appear to be somewhat less active biochemically than
either the black Aspergilli or members of the A. flavus-oryzae group. It is,
nevertheless, a vigorously growing species with definite biochemical pos-
sibilities and should not be overlooked in any program relating to mold
fermentation.

Aspergillus alliaceus appears periodically upon alliaceous bulbs and
occasionally upon cacti as at least a secondary parasite. It is not in-
frequently isolated from soils, and appears to be fairly common in the
southwestern states of Texas, New Mexico, and Arizona. It has also
been isolated from soils of other areas including Southern Mexico and
India. Nothing is known regarding its biochemical possibilities.

Only the type strains of A. avenaceus and A. panamensis are known and
neither has been shown to have any economic importance.

Chapter XIX
THE ASPERGILLUS TAMARII CxROUP

Outstanding Characters

Conidial heads radiate, generally loose-textured, hemispherical to globose,
yellow-brown to olive-brown in color with green shades entirely lacking
or only transiently produced in early stages.

Conidiophores colorless, typically roughened throughout a part or all
of their length.

Vesicles subglobose to globose, fertile over the upper half to the entire
surface.

Sterigmata in one or two series depending upon the species and strain,
often showing both conditions within the same head.

Conidia heavy-walled, rough, elliptical, pyriform or subglobose, de-
pending upon the species.

Sclerotia commonly present, purple to reddish-purple or black, white-
tipped when young.

The Aspergillus tamarii group represents a collection of more or less
closely related forms that are believed to be intermediate between the
A. wentii and A. flavus-oryzae groups. Relationship to the latter group
is unquestionable since there are intergrading forms which almost com-
pletely bridge the gap from one group to the other. The group embraces
two principal series: one, typified by A. terrieola, is characterized by dull
yellow-brown conidial heads which never show any trace of green; the
other, typified by A. tamarii, possesses dark brown conidial heads which
commonly show transient shades of olive-green during the period of rapid
growth.

Group Key
I. Conidia strongly elliptical, lemon-shaped A. citrisporus von Hohnel

II. Conidia not strongly elliptical.

A. Conidial heads light yellow-brown when mature, showing no green color at
any stage.

1. Colonies predominantly floccose, conidia with prominent projecting

tubercles A. lulcscens Bainier

2. Colonies not predominantly floccose.

a. Heads large, conidia coarsely roughened with flattened bars and

tubercles A. terrieola Marchal

b. Heads small, conidia finely roughed

A. terrieola var. americana Marchal

250

THE ASPERGILLUS TAMARII GROUP 251

B. Conidial heads dull dark brown when mature.

1. Green shades commonly evident, but confined to early stages

A. tamarii Kita

2. Green color persisting for several days, but eventually disappearing

Bronze series

Aspergillus citrisporus von Hohnel, in Sitzungsber. K. Akad. Wiss. Wien,

Math. -Naturw. Kl. Ill, I Abt., p. 987, 1902. See The

Aspergilli, Thorn and Church, pp. 191-2. 1926.

Colonies on Czapek's solution agar, at room temperature, spreading
fairly rapidly as submerged mycelium, producing a sparse aerial growth
of conidiophores only; conidial areas at first yellow then gold and finally
orange-brown (fulvus of Saccardo's Chromotaxia, approximately Mikado
brown of Ridgway); reverse colorless. Conidial heads up to 500m in
diameter, globose, radiate. Conidiophores 1 to 2 mm. long by 20 to 25m
in diameter, with walls thin, about 1m in thickness, turgid and studded
with granules when examined dry, finely roughened, often collapsing in age.
Vesicles 30 to 50m in diameter, nearly globose, fertile over the whole surface.
Sterigmata in one series, 8 to 12m by 3 to 4m, producing loosely radiating
chains of conidia, at first yellow or golden then brown. Conidia lemon-
shaped, 5 to 9m by 5 to 6m, rough from irregularly branching ridges of
coloring substance between the inner and outer walls. Sclerotia or per-
ithecia verbally reported by Thaxter but not seen by us.

Diagnosis based on Thaxter's isolate (Thorn Xo. 4181.10). Thaxter
gave no description. He obtained it several times from caterpillar dung.
It grows and fruits more abundantly on Sabouraud's agar but it is difficult
to keep viable in stock cultures. Additional collections include strains
from caterpillar dung in Ann Arbor, Michigan, and Hanover, New Hamp-
shire.

Aspergillus lutescens Bainier nomen nudum; described by Thorn and
Church, in The Aspergilli, p. 193. 1926.

Colonies upon Czapek's solution agar rapidly growing and broadly
spreading, floccose-woolly, at first white, becoming rusty-yellow as conidial
formation begins and develops unevenly over the surface (fig. 68 A),
finally becoming chestnut-brown when conidial areas are mature; reverse
of colony pale yellow. Conidial heads radiate, hemispherical to subglobose,
approximately buckthorn brown to Dresden brown (Ridgway, PI. XV),
comparatively small, ranging from 100 to 300m in diameter. Conidiophores
12 to 15m in diameter, varying greatly in length, mostly short, arising from
the substratum, or as branches of aerial hyphae with walls pale yellowish
and with pitting present but not conspicuous, not giving a rough appearance.
Vesicles globose to subglobose (fig. 68 B), 20 to 40m in diameter. Sterigmata

* 4k
ma

A

D

^S^]ri;^»^

0-'

B o

**

E

A

c

F

Fig. 68. A-C, Aspergillus lutescens NRRL No. 426: A, Colonies on Czapek's
solution agar showing extreme floccose habit and light sporulation, 10 days; B
Single conidial head, X 500; C, Conidia, X 1200. D-F, Aspergillus temcola NRRL
424: D, Single colony on Czapek's solution agar showing somewhat floccose but
heavily sporing colony, 10 days; E, Conidial heads showing sterigmata in a single
series, X 500; F, Conidia, X 1200.

252

THE ASPERGILLUS TAMARII GROUP 253

in one series in smaller and crowded heads, up to 15 to 20/x by 4.0 to 5.0m;
sterigmata often in two series in larger heads with primaries about 15 to
18m by 4 to 5m, secondaries 12 to 14m by 4 to 5m. Conidia subglobose,
varying from 5 by 7m to 8 by 9m, conspicuously roughened with prominent
tubercles of color (fig 68 C).

The species is known only in the type culture from the Bainier collection,
NRRL Xo. 425 (Thorn Xo. 4040.478), and as a second strain, XRRL Xo.
420, isolated in the Soil Microbiology Laboratory, Bureau of Plant Industry,
Washington, D. C, about 1939.

Aspergillus terricola Marchal, in Rev. Mycologique 15, Xo. 59: 101-103.

1893.

Colonies umbrinus; mycelial hyphae 3 to 5m in diameter, without anasto-
moses; conidiophores hyaline, continuous or septate in age, 000 to 1,000m
by 7 to 10m (whole depth of colony growth); vesicles subglobose, hyaline,
39 to 50m, radiately covered with sterigmata; sterigmata in one series, 12
to 15m by 4 to 7m; conidia umber (Sacc), ovate or elliptical then globose,
rough, with colorless connectives.

Description, from Marchal, of a culture isolated from soil in Belgium;
not reported elsewhere, see variety below.

Aspergillus terricola var. americana Marchal, cultural description by Thorn
and Church, in Am. Jour. Bot. 8: 125. 1921.

Colonies on Czapek's solution agar growing rapidly at room temperature,
often somewhat floccose in central colony areas (fig. 68 D), ranging from
shades near yellow ochre (Ridgway, PI. XV) when young, to Dresden brown
or mummy brown in age, near Saccardo's umbrinus (PI. VII A); aerial
growth largely consisting of crowded conidiophores; reverse uncolored.
Conidial heads radiate, hemispherical to subglobose, loose in texture,
consisting of comparatively few divergent chains of conidia, up to 200m in
diameter. Conidiophores 300 to 000m in length by 0 to 8m in diameter,
with walls pitted. Vesicles globose to subglobose (fig. 08 E), up to 25m
in diameter, fertile over the upper two-thirds or three-fourths. Sterigmata
in one series, 7 to 10m by 2 to 4m- Conidia tuberculate (fig. 08 F) from the
presence of color bars variously distributed between the outer and inner
wall, ovate to nearly globose, from 3 by 5m up to 5 by 7m, usually about
5.5m, occasionally 5 to 8m in diameter.

Type culture XRRL Xo. 424 (Thorn Xo. 4838) isolated by F. M. Scales
from redland soil in Georgia and discussed by Scales, in Jour. Biol. Chem.
19: 459-472, 1914, under the name A. terricola Marchal. Scales' culture
was submitted to Marchal, who designated the form as A. terricola var.
americana Marchal, distinguished as follows: "The dimensions of the

254

A MANUAL OF THE ASPERGILLI

vesicles 14 to 20m instead of 30 to 50m; of the sterigmata 5.6 to 10.5m by
2.2m instead of 12 to 1 5m by 4 to 7m ; the spores only very delicately verrucose,
separate your fungus from A. terricola."

Fig. 69. Aspergillus tamarii. A, Colony growing on Czapek's solution agar
characterized by heavy conidial production, strain XRRL No. 427, 10 days. B,
Conidial heads, X 160. C, Single head, X 300. D, Single head further enlarged
showing rough walls of conidiophore, and sterigmata in two series, X 500.

Aspergillus tamarii Kita, in Centralb. f. Bakt. etc. 2 Abt., 37, No. 17/21,
pp. 433-452. 1913. .Characterization of .4. tamarii Kita given by
Thorn and Church in Am. Jour. Bot. 8: 118. 1921. See also Thorn
and Church, The Aspergilli, p. 194. 1926.

Colonies on Czapek's solution agar spreading broadly at room tempera-
ture (fig. 69 A), with vegetative hyphae mostly submerged, fruiting areas

THE ASPERGILLUS TAMARII GROUP 255

at first colorless, then passing through orange-yellow shades to brown in
old colonies, variously Isabella color, light brownish-olive, buffy-citrine,
medal bronze, or raw umber (PI. VII B) (Ridgway, Pis. XXX, XVI, and
IV, Column 19, and PI. Ill, Column 17); not showing true green, but often
presenting a suggestion of green that is transient and limited to areas of
young heads; reverse uncolored or occasionally pinkish. Conidial heads
varying greatly in size in the same fruiting area, from more or less columnar
to nearly, but not completely, globose and up to 300m in diameter, with
radiating chains and columns of conidia. Conidiophores arising from
submerged hyphae, up to 1 to 2 mm. in length, colorless, with walls be-
coming abruptly thinner at the base of the vesicle, frequently showing
irregular thickenings within, as a rule markedly rough or pitted throughout
part or all of their length (fig. 69 D), sometimes appearing smooth or nearly
so when examined in liquid mounts, but consistently rough or pitted when
examined dry. Vesicles globose to subglobose, 25 to 50m in diameter
(fig. 69 C), with fairly thin walls which frequently crush in mounts, fertile
over almost the entire surface. Sterigmata, in one series in small heads,
in two series in large heads; primary sterigmata commonly 7 to 10m by 3 to
4m, becoming 20 to 35m long in gigantic heads, secondary sterigmata 7 to
10m by 3m- Conidia ranging from more or less pyriform, through sub-
globose to globose, conspicuously roughened from prominent tubercles and
bars of orange-3rellow coloring matter deposited between the loose outer
wall and the firm inner wall, commonly ranging from 5.0 to 6.5m in diameter,
occasionally up to 8m- Sclerotia produced by many strains, usually purplish
or reddish-purple, globose to pjniform with apex white.

Species characterization is based upon Thorn and Church's culture No.
4235.12 (NRRL No. 429) which was submitted to and identified by Kita
as A. tamarii (see Thorn and Church, Am. Jour. Bot. 8 : 118. 1921).

The organism described by Kita proved to be one of a great series repre-
sented in our collection. It is common among cultures examined from
North and South America, from Japan, China, India, and from Europe.
The species has been found to be quite common in soil collected from
many areas in the United States.

The outstanding characters are orange-yellow to brown colonies; coarse,
colorless conidiophores, usually roughened but with this character sometimes
obscure; large, radiate, loose-textured, conidial heads; sterigmata in one or
two series, commonly with single and double sterigmata in the same head;
conidia more or less pyriform 5 to 8m in long axis with tubercles or bars of
orange-yellow coloring matter between the inner and the outer cell walls.

In preparing the manuscript for "The Aspergilli" (1926), Thorn and
Church introduced into their general key, without name, on page 248
under No. 279, a series of forms with morphology and general .appearance

256 A MANUAL OF THE ASPERGILLI

bridging the gap between .4 . tamarii Kita and the A . flams group. Without
publication they referred to these as "The Bronze Series." These strains
have the yellow-green color of A. flavus during the early stages of their
development, but subsequently develop the yellow to brown colors of A.
tamarii. Recognition of this border group is necessary since some strains
of A. tamarii do not assume a definitely green color at any state in their
development, while others show green as a transient character. Strains
of .4. flavus, on the other hand, are typically characterized by the green
to yellow-green colors. Furthermore, the conidia of A . tamarii are typically
quite roughened, showing prominent tubercles or bars of coloring matter
deposited between the outer and inner walls. In contrast, the conidia of
A. flavus are less coarsely roughened and show more numerous and smaller
tubercles or echinulations, as well as a greater tendency for the coloring
substance to be generally diffused throughout the spore envelope. The
forms under consideration show, in some degree, the coloration and spore
characters of both groups and are believed to be truly intermediate be-
tween A. tamarii and A. flavus.

The fact that we received from Baarn in 1933, as Blochwitz's A. luteo-
virescens Bloch. (Ann. Mycol. 31: 73-83. 1933) a culture (Thorn No.
5345) which represented satisfactorily this intermediate series is not
accepted as justifying the assignment of this name to the series, since the
morphological characters displayed by the strain were so completely at
variance with the original description, and since Blochwitz considered his
species to be close to A. ustus. We question whether any sharp line of
separation can be drawn between the two series because of the repeated
appearance of intermediate forms. While we do not feel justified in
assigning to these forms any specific designation, we do feel obligated to
continue to call attention to their existence. We have at times considered
the desirability of moving the whole A. tamarii complex over into the A.
flavus-oryzae group, but this course has been abandoned since it was felt
that to do so would introduce into an otherwise perfectly integrated
group, a series of organisms whose relationship to them, while strongly
suggested, is not proved, and which in its typical form would introduce
discordant features.

The species listed below are believed to represent probable synonyms:

Biourge attached the manuscript name A. vulpinus to a member of the A. tamarii
series (Thorn No. 4733.146) and contributed it to our collection, but it does not seem
sufficiently different from the species to warrant separation.

One strain of A. tamarii was found in the Bainier collection (Thorn No. 4640.397)
as A. cacao, nomen nudum; another under the same name came from Pribram.

A. gigas Spegazzini, Myc. Argent. V, in An. Mus. Nac. Buenos Aires Ser. B. Tome
13: 424. 1911, was described from decaying coffee leaves in terms that suggest its
relationship to A. tamarii.

THE ASPERGILLUS TAMARII GROUP 257

A. spadix Amons, in Archief voor de Suikerindustrie in Nederlandsch-Indie Jaarg.
29, Deel 1, pp. 12-14. Jan.-June 1921. From the description this is a synonym of
A. tamarii Kita. Colonies described as yellow-brown to deep brown, growing well
one ommon laboratory media, without aerial mycelium; in reverse colorless; rice-
colored to light violet at first, then light fuscous brown; conidiophores up to 2 to 3
mm. by 8 to 9/x with walls about 0.9m thick and pitted or rough; vesicles globose, up
to 50m in diameter, or almost clavate in small heads; conidia 5.5 to 7.2m, rough.

Culture: Amons. Not studied by us.

A. erythrocephalus B. and C, in Jour. Linn. Soc. (London), Bot. 10: 362. 1869.
(See Fungi Cubensis Wrightiana, 1868; Type No. 642 in Curtis Herbarium de-
posited in the Cryptogamic Herbarium of Harvard University, bears Wright's No.
764. Part of the original material was removed by Dr. Farlow and given to Thorn
for study.)

Microscopic examination of this type specimen gives measurements as follows:
conidiophores 45 to 70m in diameter, up to 2 mm. in length, with walls very heavy 5 to
12m thick, varying from 5 to 6m in the broader part to 10 to 12m at the narrower base,
pitted or roughened; vesicles up to 100m in diameter, nearly globose, fertile all over;
head washed free from spores about 150m in diameter; sterigmata in two series, pri-
mary 8 to 10m in length, secondary 8 to 9m in length ; conidia commonly 8 by 6m, ranging
up to 8 to 12m by 5 to 9m, finely pitted or roughened with rather thin walls. Colors in
the material are questionable on account of the age of the collection.

Cultures: None. Type material only known. Placed between the A. tamarii and
A.flavus groups. The amended description is offered due to the existence of a type
specimen with very conspicuous characters under a name only very briefly described
in 1869. When grown upon natural substrata such as grains, et cetera, conidiophores
and heads of A. tamarii become very much larger than those ordinarily produced in
culture media. This might account for this specimen which bears the name A.
erythrocephalus B. and C.

Occurrence and Economic Importance

Of the species included in this group of brown-spored Aspergilli, only
A. tamarii is in any sense widely distributed or common in nature. Asper-
gillus lutescens is known only as the type culture and as a second isolation
made in Washington, D. C, many years later. Aspergillus terricola has
not been positively identified since its description, although the form with
smaller heads and less coarsely roughened spores designated A. terricola
var. americana by Marchal is occasionally encountered. Aspergillus
tamarii is, however, a cosmopolitan mold upon vegetable material under-
going slow decomposition and can be isolated from almost all soils examined.
Like A. niger and A.flavus, it is more frequently recovered from warm and
semi-tropical soils than from cool, temperate soils, although it occurs in
the latter. The species commonly appears with A. flavus and A. oryzae
as a constituent part of the "koji" used in the fermentation industries of
the Orient. Certain strains apparently produce appreciable amounts of
diastatic and proteolytic enzyme, while other strains are known to produce

258 A MANUAL OF THE ASPERGILLI

kojic acid. Gould reported this in 1938, and it was subsequently confirmed
by A. J. Moyer (unpublished notes) for a strain isolated from Panama
soil at the Northern Regional Research Laboratory in 1941.

Kita's culture was isolated from a soybean sauce termed "Tamari",
hence the species name. Tamari is made by a shorter fermentation process
than soy sauce or shoyu, and differs from it in flavor. Ivita believed that
where it was made empirically, it owed its individuality to the particular
aspergillus which he isolated and described.

Chapter XX
THE ASPERGILLUS FLAVUS-ORYZAE GROUP

Outstanding Characters

Colonies varying from very light greenish-yellow to deep yellow-green

(Ivy Green).
Conidiophores rough or pitted, colorless.
Heads hemispherical to columnar to subglobosc.
Sterigmata in one or two series, often varying in the same head.
Vesicles variable in form, from hemispherical to dome-shaped in small

heads to globose in large heads.
Conidia more or less roughened, varying in color as the colony.
Sclerotia characteristic of many strains, generally grayish-brown to

black, entirely lacking in others.

Two species names are widely used for members of this cosmopolitan
group. Aspergillus oryzae is applied quite generally, without regard to
morphology, to the strains used by the Japanese and Chinese in the fermen-
tation of rice and soy products. Although purified cultures are used in
many places, the nomenclature is based more upon utilization than upon
morphology. There appears, however, in these industries, a series of strains
with long conidiophores, radiate heads, mostly greenish-yellow, with the
green often fading completely in old cultures. These strains appear to be
most commonly used in the production of the diastatic type of ferments and
to be distributed in the great culture collections as Aspergillus oryzae
(Ahlb.) Cohn. Such strains seem to be mostly oriental or tropical in origin.
Strains with shorter conidiophores and yellowish-green heads, on the other
hand, appear wherever fermenting or decaying materials are examined
microscopically, or by culture. Aspergillus flavus Link has been accepted
as a species aggregate for this second array of forms from which the segrega-
tion of sections for description as separate species has been found difficult,
if not almost impossible. If one wishes to perpetuate species names as
roughly covering aggregates of closely related but varying strains, bearing
always in mind that no sharp lines of differentiation exist, certain applica-
tions of names may be made arbitrarily about as follows:

Group Key

I. Sterigmata mostly in one series, double sterigmata also present.

A. Conidiophores long, 1-several mm., heads radiate, greenish-yellow; conidia
pyriform, more or less roughened, variable in size up to 6, 8, or even 10ju
in long axis A . oryzae ( Ahlburg) Cohn

259

260 A MANUAL OF THE ASPERGILLI

B. Conidiophores 600 to 1700m; heads radiate, hemispherical, pale greenish-

yellow; conidia smooth, globose 3.0 to 4.6^ A. micro -virido-citrinus

Costantin and Lucet

C. Conidiophores mostly less than 500/x; heads deep yellowish-green (Ivy

Green); described as a parasite of the mealy bug of cane, occasionally
elsewhere A . parasiticus Speare

II. Sterigmata mostly in two series but single series common and often in same head,
small heads usually showing single series only.

A. Conidiophores very variable in length, mostly 400 to 1000m; heads in various

yellowish-green shades A. flavus Link

B. Conidiophores mostly borne as short branches from trailing hyphae forming

an uneven cottony mass; heads white to yellow with traces of green only

A. effusus Tiraboschi

Literally hundreds of strains of this group have been collected and
compared. Many of them have been isolated from fermentation investiga-
tions in the laboratory and from industrial processes. No correlation of
colony appearance, conidiophore or head morphology, color or microscopic
detail, with actual utilization has been proved. A culture labeled A.
oryzae (Ahlburg) Cohn, NRRL Xo. 447 (Thorn No. 113) has been preserved
for over 30 years without apparent change in morphology. It is probably
derived from Cohn's organism. When, however, we scrutinize the Asper-
gilli obtained from the rice or soy fermentations of the Orient, cultures of
the type represented by this strain are not the most common. The pre-
eminently useful strains usually have the aspect of forms intermediate
between A. flavus and A . oryzae. The dwarf green A . parasiticus of Speare
isolated from dead mealy bugs of sugar cane in Hawaii proved no more
parasitic to the same species of insects in the Barbados than other .4 . flavus
strains sent with it. Teizo Takahashi contributed his series of strains under
the letters used in his publication (1913). These are discussed at some
length in Thorn and Church's paper on A. flavus, A. oryzae and associated
species (1921), and also in "The Aspergilli" (192G, p. 202). It is sufficient
to say that they vary all the way from almost white with few lightly colored
heads to rich yellow-green in which heads are very numerous and fairly dark.
They vary likewise in the length and diameter of their conidiophores.
Characters of color and conidiophore length are not always correlated,
although it is generally true that the darker conidial masses are borne upon
shorter stalks. The collections contributed by Oshima, Kita, Hanzawa,
and others from Japan as well as those isolated from commercial "Koji''
(sold as inoculum for fermentation industries) showed mainly the .4 . flavus
morphology. Strains of this series appear constantly where cultures are
made from soil or from decaying vegetation. A . flavus and its allies appear
in collections from every correspondent who contributes Aspergilli. It is
debatable whether the worker will be benefited or confused by the introduc-

THE ASPERGILLUS FLAVUS-ORYZAE GROUP 261

tion of some of the species names applied to members of the group. It
must not be forgotten that any variant from the dwarf and deep green A.
parasiticus to the longest stalked and palest greenish-yellow .4. oryzac may
be found if we look for it.

Aspergillus oryzae (Ahlburg) Cohn, in Jahresb. Schles. Gesell. Vaterl.
Cultur (1183) 61: 226 Breslau. 1884.

Synonym: Eurotium oryzae Ahlb. The name E. oryzae with an incom-
plete description for the sake organism was published by
Korschelt, in Dingier 's Polytechnisches Jour. 230: 330.
1878, as taken from a letter from "Herr Ahlburg." See also
Thorn and Church, Amer. Jour. Bot. Bot. 8: 106. 1921, and
The Aspergilli, p. 198. 1926.
Colonies on Czapek's solution agar rapidly spreading with vegetative
hyphae mostly submerged and forming a white to gray mycelial layer in the
form of a tough felty mass (fig. 70 A); developing pale greenish-yellow
shades with the production of ripening conidial areas, varying from lime
green to mignonette green (Ridgway, PI. XXXI, column 25) with the green
disappearing later and the general color shifting to yellowish-brown shades;
mycelium and agar uncolored. Conidial heads predominantly large, abun-
dant, globose, radiate, with chains of conidia separate rather than adhering
(fig. 70 D), giving the pale yellow shades of the colonies. Conidiophores 2
to several mm. long by up to 20 to 25m in diameter with walls rather thin,
definitely pitted or rough (fig. 70 F), colorless. Vesicles globose to sub-
globose, less often hemispherical, up to 50 or even 70m with walls 1 to 1.5m-
Sterigmata commonly in one series up to 15 or 20m long by 3 to 5m; or in two
series with primary sterigmata up to 12 by 5m, and secondary sterigmata 10
to 12m by 3.5m (fig. 70 B). Conidia more or less pyriform (fig. 70 F), vary-
ing greatly in size in the same culture and in different strains, 3 by 4m, 4 by
5m, 5 by u> or up to 9m or 10m hi long axis occasionally, rather thin-walled,
roughened, becoming coarsely and deeply roughened in some strains.
Sclerotia dark, few and not forming clumps, produced sporadically under
undefined conditions.

Diagnosis based primarily on culture XRRL Xo. 447 (Thorn Xo. 113)
received from the Centraalbureau at Baarn and believed to be derived from
Cohn's original strain.

While the above description is believed to conform closely to the original
conception of .4. oryzae, strains possessing the essential morphology de-
scribed but which are heavier sporing and somewhat darker in color are more
commonly encountered. Culture XRRL Xo. 458, obtained from Dr.
Oshima as strain AoOld and shown in PI. VII C, and Fig. 70 C-F, is repre-
sentative of these forms.

CP

^n1

mm'"7 9 ^Sm \

*£$*§!

-•^j

«

%

■

i

F

Fig. 70. Aspergillus oryzae. A and B, Strain XRRL Xo. 447 (Thorn No. 113):
A, Single colony on Czapek's solution agar, loose-textured, conidiophores long and
limited in number, 10 days; B, Details of single head, vesicle thin-walled, sterigmata
mostly in two series, X 480. C-F, Aspergillus oryzae XRRL Xo. 458: C, Single
colony on Czapek's solution agar, comparatively heavy sporing, 10 days; D, Conidial
heads of the same, X 18; E, Conidial heads further enlarged, X 270; F, Single head
showing vesicle, sterigmata in a single series, and roughened conidiophore, X 775.

262

THE ASPERGILLUS FLAVUS-ORYZAE GROUP 263

The production of perithecia by members of this series was reported by
Bezssonoff (1919) without adequate description and by Zikes (1922) whose
culture, as received from him, belonged in the A. glaucus group. No
ascosporic form is verifiable for the group thus far.

Aspergillus micro-virido-citrinus Costantin and Lucet, in Ann. Sci. Xat.

Bot. (IX) 2: 158. 1905.

The appearance of colonies and measurements of conidiophores, heads,
and spores indicate a form intermediate between A. flavus and .4. oryzae
except for its small conidia. The description is very nearly satisfied by
Takahashi's culture "P" (XRRL Xo. 480). It was found to grow between
15° and 45° C. and to be pathogenic to rabbits. Colonies were greenish-
yellow to predominantly yellow but contained some definitely green admix-
ture in contrast to A. oryzae, which often lacks green color entirely. Co-
nidiophores 600 to 1700m in length, up to 21m in diameter near the vesicle,
uncolored, "granular" (= pitted) above, smooth toward the base. Vesicles
24 to 62m in diameter. Sterigmata varying in size and arrangement with
the size of the heads examined. Conidia globose, smooth, 3 to 4.6m (3.1m
as a minimum to occasional diameters of 5.5m).

An occasional culture shows the morphological characters described by
Costantin and Lucet. Xo actual identity has been proved.

Aspergillus flavus Link, in Obs. p. 16. 1809; also in Sp. Plant. 6: 66.
1824, cited as synonym of Monilia flava Persoon, Myc. 1, p. 30.

Synonym: Eurotium Aspergillus flavus DeBary and Woronin, in Beitrage
zur Morphologic und Physiologie der Pilze, III Reihe, p.
380. 1870. Exsiccati by Brefeld preserved in Rabenhorst,
Fungi Europaei Edit. Xov. ser. II, Xo. 2135; one packet
in the collection of the Xew York Botanical Garden.

Colonies on Czapek's solution agar spreading rapidly, with floccosity
limited to scanty growth of sterile hyphae in older and dryer areas among
crowded conidiophores; conidial areas range in color in various strains from
sea-foam yellow through chartreuse yellow, citron green, lime green, to
Kronberg's green (PI. VII D and fig. 72 A), or even to ivy green, (See Ridg-
way, PI. XXXI, column 25), yellow-green colors are either persistent or, in
old colonies, altered by the disappearance of the green factor leaving shades
of yellow-brown; reverse yellowish at first, passing over into brown shades
in age. Conidial heads vary from small with a few chains of conidia to
large radiate (fig. 72 E) or columnar masses in the same culture and varying
mixtures of different types and sizes of head. Conidiophores mostly arising
from submerged hyphae, commonly 400 to 1000m l°ng by 5 to 15m in diame-

264

A MANUAL OF THE ASPERGILLI

Br ^-. <S*

§

Ik

3§

\^-^^

S2C

\r — v^

Hi*"*

Jr- — f^ffe

\\vS?3@

1 k r\ NV r-*K '^^

to

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e©

Fig. 71. Conidial structures in the Aspergillus flarus-oryzae group. A, A. flavus,
NRRL No. 482: A1; typical, large, radiate to globose head showing sterigmata in two
series; A2, small, loosely columnar head showing single series of sterigmata. B, A.
effusus, NRRL No. 506: Bu large radiate head showing double series of sterigmata;
B2, smaller head showing sterigmata in a single series; B3, diminutive head borne
upon one of a chain of foot cells. In this group single and double sterigmata often
occur in the same head (not illustrated).

Fig. 72. Aspergillus flavus. A, Typical strain, NRRL No. 1957, showing crowded
conidial heads and occasional black sclerotia. B, Heavy sclerotium producing
strain, NRRL No. 500. C, Strain used for the production of kojic acid, NRRL No.
484 (This approaches the character of A. oryzae). D, Dr. White's strain of A. flavus
(NRRL No. 501) showing characteristic thin growth and sparse sporulation. E,
Typical conidial heads, X 18. F, Single head showing vesicle, sterigmata, conidia,
and the roughened conidiophore, X 780. All cultures on Czapek's solution agar,
10 days.

265

266 A MANUAL OF THE ASPERGILLI

ter, with walls pitted, rough (fig. 72 F) almost spiny in appearance, broaden-
ing upward and gradually enlarging into vesicles 10 to 30 or 40m in diameter,
dome-like in the smaller heads, flask-shaped in larger heads (fig. 72 F).
Sterigmata in a single series in many smaller heads (fig. 71 A2), or both single
and double series on the same vesicles in large heads (fig. 71 Ai), varying
from single sterigmata only 10 to 15m by 3 to 5m, to primary sterigmata 7 to
10m by 3 to 4m and a secondary series 7 to 10m by 2.5 to 3m- Conidia pyri-
form to almost globose, nearly colorless to definitely yellowish-green, vary-
ing from 3m, 3 by 4m, 4 by 5m, or even larger and marked variously with pits,
echinulations, or irregularly winding color bars and ridges to give a rough-
ened effect of varying intensity. Sclerotia, when found, at first white then
brown, hard parenchymatous, and a few strains white tipped, produced by
some strains regularly and abundantly (fig. 72 B), scantily by others under
undefined conditions. Perithecia not found.

Description originally based upon culture NRRL No. 482 (Thorn No. 108)
from the Centraalbureau at Baarn, Holland, but supplemented by obser-
vation of many hundreds of cultures from many substrata and all parts of
the world.

Unless segregation under a specific name is supported by adequate
morphological and reproducible cultural data, there is no way to identify
the organisms intended. Applying these criteria, no reasons are seen for
the use of the following specific designations:

A. wehmeri Constantin and Lucet, in Ann. Sci. Nat. Bot. (IX) 2: 162. 1905.

A. variabilis Gasperini, in Atti. Soc. Toscana Nat. Sci. Pisa Mon. 8, fasc. 2: 326.
1887.

A. pseudo-flavus Saito, in Centralb. Bakt. etc., 2 Abt., 18, No. 1/2, p. 34, figs. 15-18.
1907, or its synonym S. pseudc-flava (Saito) Sacc, in Syll. 22: 1260. 1913.

A. siebenmanni Constantin and Lucet, in Ann. Sci. Nat. Bot. (IX) 2: 162. 1905,
is a bibliographic species based upon an organism isolated from the human ear and
diagnosed by Siebenmann (Zeitsch. f. Ohrenheilk 12: 1883) as A. flavus. The de-
scribes regarded it as a separate species based only upon the description given by
Siebenmann.

A. gymnosardae Yukawa, in Jour. Col. Imp. Univ. Tokoyo 1: 362, PI. 18, figs. 1-7.
1911. A member of the A. flavus-oryzae group with measurements intermediate
between more typical representatives of the two species.

A. Ihomii Graff, nomen nudum, a heavy sclerotium-producing strain distributed
by Graff but never described. No diagnostic basis for the name was presented.

A. pollinis Howard, in Am. Bee Jour. 36: 577-578. 1896, was discussed as an or-
ganism causing "pickled brood and bee paralysis" (See also idem. 38: 530-531. 1898).
Turesson (Svensk Bot. Tidskr. 11: 30. 1917) decided the mold was A. flavus.

Aspergillus parasiticus Speare, in Hawaiian Sugar Planters' Exp. Sta.,

Path, and Physiol. Ser. Bui. 12, p. 38, pi. 3-4. 1912. See

Thorn and Church, The Aspergilli, p. 203. 1926.

Colonies on Czapek's solution agar with sucrose spreading rapidly,
forming a surface growth of crowded conidiophores with very few sterile

THE ASPERGILLUS FLAVUS-ORYZAE GROUP 267

hyphae (fig. 73 A), in deeper yellow-green shades near ivy green (Ridgway,
PI. XXXI); reverse uncolored or yellowish. Conidial heads radiate,
abundantly produced and giving color to the colony. Conidiophores given
by Speare as 300 to 700m long, commonly under 400ju, with walls colorless,
prominently rough or pitted, enlarging from 3/x at the foot up to 10 to 12/x,
and passing into vesicles up to 35/* in diameter (fig. 73 B). Sterigmata in
one series, 7 to 9/x by 2.5 to 3/x, closely packed over the vesicular surface,
yellow. Conidia pyriform to globose, very rough, 4 to 5/x, occasionally 6/x
in long axis, green. Xo sclerotia or perithecia reported.

Described as parasitic upon the mealy bug of sugar cane (Pseudococcus
calceolariac Mask.) in Hawaii. Type culture XRRL Xo. 502 (Thorn No.
3509) received from Speare. Cultures with the same morphology Avere
isolated from infected mealy bugs from Demerara by Thom, and in Louisi-
ana by Kopeloff . Johnston, working in Puerto Rico, considered that he had
proved infectivity to be a strain function among organisms of the A. flavus
series rather than associated with morphology. Blochwitz (Ann. Mycol.
32(1/2) : 8G. 1934) has called another nearly allied form A. flavus vsuwiridis,
but gives no adequate data for separation. Cultures with these characters
are occasionally obtained from sources not known to be associated with
disease of insects. Shih has likewise described from China as Aspergillus
chungii (Lingnan Sci. Jour. 15(3): 378. 1933) a strain which apparently
duplicates .4. parasiticus.

Aspergillus effusus Tiraboschi, in Ann. di Bot. (Rome) 7: 16, fasc. 1.

1908. See also Thom and Church, in Am. Jour. Bot. 8: 109-110.

1921, and Thom and Church, in The Aspergilli, p. 208. 1926.

Colonies on Czapek's solution agar rapidly and broadly spreading, floc-
cose or piled cottony white (fig. 73 C), becoming dirty yellowish or, in re-
stricted areas, pale greenish-yellow, then passing over into dull buff or tan
shades as heads mature ; reverse and agar yellowish. Conidial heads usually
more or less columnar, mostly small, a few of them fairly large, many of
them upon short conidiophores (fig. 71 B3 and 73 E), often less than 100/x
long and 5 to 10/x in diameter, arising from the trailing floccose hyphae,
quickly losing their yellow-green color. Conidiophores with walls pitted or
roughened, sometimes bearing granules (produced by drying droplets of
exuded fluid). Vesicles mostly under 20/x in diameter (fig. 71 B2). Sterig-
mata in one series in small heads, in either one or two series in large heads
(fig. 71 BO, approximating the A. flavus type. Conidia pyriform to
globose varying from 3 by 4/x to 5 by 7m- Xo sclerotia or perithecia re-
ported. The species was described originally from rotten corn (Zea
Maijs).

Culture description as given centers around culture XRRL No. 506
(Thom Xo. 130) isolated by Dr. B. F. Lutman, Burlington, Vermont. Other

T*iJ&

B

•

i ^|

' .if--

0&

A

■

00

V

■

v.
I

Fig. 73. A and 5, Aspergillus parasiticus, NRRL No. 465: A, Colony on Czapeks'
solution agar showing heavy production of short-stalked conidial structures, 10
days; B, Single conidial head showing single series of sterigmata and roughened
conidiophore, X 500. C-F , Aspergillus effusus, NRRL No. 506: C, Colony onCzapek's
solution agar showing characterisitc floccose habit and light sporulation, 10 days;

D, Conidial structures arising from aerial hyphae characteristic of species, X 165;

E, Single short-stalked fruiting structure arising from aerial hyphae, X 300; F,
Conidial head-Showing sterigmata mostly in two series, roughened surface of conidio-
phore not apparent, X 500.

268

THE ASPERGILLUS FLAVUS-ORYZAE GROUP 269

strains studied include isolations from cornmeal from Indiana and from
mealy bugs in Puerto Rico. Superficially this species bears little relation
to A. flavus. Detailed microscopic examination of heads and spores,
however, reveals true and close relationships. Selective transfer from
original colonies permitted changes in the predominance of the sterile
areas over the fruiting areas without changing the general habit or nature
of the colony. Blochwitz (Bot. Centralb. Beiheft Abt. Anat. Phys. 48:
176-182. 1931) regarded A. effusus as a floccose type of A. flavus. This
position can be supported, but the authors feel that the species should be
maintained since it is strikingly different from A. flavus in its general colony
appearance, and since it is repeatedly, although infrequently, isolated from
nature.

Whether Sterigmatocijslis lulea Bainier (Bui. Soc. Bot. France 27: 27. 1880)
was one of these can only be guessed from culture NRRL No. 508 (Thorn No.
4640.473) received from the Bainier collection under this name. The strain is close
to A. effusus. Bainier did not claim identity with S. lutea van Tieghem (Bui. Soc.
Bot. France 24: 103. 1877), which was entirely undescribed.

Aspergillus jeanselmei Ota, in Ann. de Parasit. 1(2): 137-146. 1923, as received
from Baarn in 1939 (NRRL No. 507: Thorn No. 5665) represented a member of the
A. flavus series with close affinities to A. effusus.

Occurrence

Members of the A . flavus-oryzae group are among the most abundant of
all the Aspergilli. They are world-wide in distribution and are omnivorous
in the substrata upon which they are able to grow and develop. They have
been isolated from the widest variety of sources including : the fermentation
industries of the Orient, grains and cereal products from different parts of
the United States, various types of forage, egg noodles, bread and other
bakery products, leather goods, dried dates, cured meats, dairy products,
nut meats, soy sauce, home-canned fruits and vegetables, textiles, paper
pulps, insects, tannin inoculum, feces, sputum, the lung of a bird, and from
the duodenum of man. They are very abundant in soil, and have been
observed in almost all samples examined. They appear to be particularly
common in the warm soils from tropical and sub-tropical areas. They vary
greatly in cultural appearance and in the detailed measurements of their
fruiting structures, and to a limited degree these differences can be corre-
lated with the sources from which they are obtained. Soil isolates com-
monly show conidial heads near yellow-green in color which are borne upon
comparatively short conidiophores. Isolates from the rice and soy fermen-
tations of the Orient often show conidial heads pale yellow-green in color
that are borne upon long, thin-walled conidiophores. Exceptions to this
very general statement are common.

270 A MANUAL OF THE ASPERGILLI

Kojic Acid

The ability of members of the A. flavus-oryzae group to produce kojic
acid has been recognized for more than three decades. It is only within the
past fifteen years, however, that serious attention has been given to this
fermentation. Beginning with the work of Challenger, Klein, and Walker
in 1929 and 1931, and continuing with that of May, Herrick, Moyer, Ward,
and Wells in 1931 and 1932, the proper nutrients and cultural conditions
necessary for its production were defined. Subsequent contributions have
been made by Kluyver and Perquin (1933) and by Barham and Smits
(1936). In all of the early reports the responsible cultures were cited as A.
oryzae, whereas in more recent ones the cultures employed have generally
been identified as Aspergillus flavus. It is of interest to note that the strain
studied by May and associates (NRRL No. 484: Thorn No. 3538) was a
thoroughly typical A . flavus when first isolated by Thorn in 1914, but during
the long period that it has been maintained in artificial culture it has gradu-
ally changed until today it more nearly resembles A . oryzae in its general
habit and coloration (fig. 72 C). Its capacity to produce kojic acid remains
undiminished, however. The culture employed by Barham (NRRL No.
625) likewise fails to satisfy the typical cultural picture of A . flavus, although
it is discussed under this name. In contrast to these cultures, other strains
belonging to this group have been under continuous laboratory cultivation
for more than 30 years without apparent change in appearance or behavior.
The above and additional references to the kojic acid fermentation are pre-
sented in the Topical Bibliography, pp. 297-298.

Enzymes

Members of the A . flavus-oryzae group produce diastatic and proteolytic
enzymes abundantly. For this reason they have been much studied, and
an extensive literature regarding mold enzymes has developed around the
use of these fungi. In large measure the alcoholic and soy food industries
of the Far East are based upon these molds and their enzymes. In the
production of alcoholic beverages, the diastatic enzymes produced by an
Aspergillus (regularly identified as A. oryzae) are employed to hydrolyze
the rice starch. Alcohol is then produced from the resultant sugars by the
addition of a fermentative yeast. In the soy industries, closely related
molds, or even the same strains, are used as a source of proteolytic enzymes.
In 1894 Takamine secured a series of U. S. patents covering the production
of diastatic enzymes and the making of alcoholic liquors (see Topical
Bibliography, p. 302). Subsequent to this, other investigators, mostly
Japanese, published a number of papers in this field. Oshima in 1922 and
1928 reported on the production of protease by members of the A. flavus-

THE ASPERGILLUS FLAVUS-ORYZAE GROUP 271

oryzae group. Today considerable quantities of diastatic enzymes, proteo-
lytic enzymes, and mixed diastatic and proteolytic preparations are being
manufactured from these molds for use in the textile and tanning industries
particularly. Within recent years considerable attention has been given
to "moldy bran" (bran seeded with selected strains of A. oryzae) as a pos-
sible substitute for malted barley as a saccharifying agent in the production
of industrial alcohol '(Underkofler, Fulmer, and Schoene, 1939; Schoene,
Fulmer, and Underkofler, 1940; Hao, 1942; Hao, Fulmer, and Underkofler,
1943; Christensen, 1943).

References to papers dealing with the production of enzymes by molds
belonging to this group are presented in the Topical Bibliography, pp.
302-304. No attempt has been made to present a complete bibliography
of the subject, but it is believed that sufficient citations are listed to intro-
duce the reader to the extensive literature of the field.

Pathogenesis (See Topical Bibliography pp. 307-310)

Pathogenesis has been occasionally reported for strains identified as mem-
bers of the A. flavus series. Observations reporting their presence in the
external ear go back to Siebenmann (1882). Ota described A. jeanselmei
as a parasite of human nails in Paris in 1923. Bereston and Keil (1941)
described a case of infected nails in which the strain, as seen by us, proved
to be a variant of A . flavus. There are ample records to show an occasional
infection of the human being. There are no data as to the route of infection,
and the question whether the parasite is a primary or a secondary (wound)
parasite stands unanswered. The constant presence of members of this
group in every human environment, together with a lack of evidence of
ability to penetrate sound human tissue, leaves some doubt — not as to its
ability to grow when once established, but whether it can actually "break
and enter" as a direct agent of disease. Aspergillus flavus is commonly
isolated from sputum. In birds, cases of lung involvement have been re-
ported. There is no question but that the mold can persist for reasonable
periods of time within the animal body. A. flavus is one of the more com-
mon air-borne molds, and occasional allergic reactions are attributed to it,
although instances where it is the sole responsible agent are not known to
have been reported.

Antibiosis

The production of antibacterial substances by strains of Aspergillus flavus
has been observed by a number of workers during the past five years.
White (1940) reported a culture of A. flavus (found by the writers to be a
somewhat atypical strain) to produce some substance which was definitely
bactericidal against some gram-negative and some gram-positive bacteria.

272 A MANUAL OF THE ASPERGILLI

A more detailed study of this strain was subsequently made by White and
Hill (1943) and the name "aspergillic acid" was assigned to the active
substance. The compound shows a comparatively high toxicity to labora-
tory animals. Utilizing the White strain, Jones, Rake, and Hamre (1943)
have made additional studies on the biological properties of aspergillic acid.
In the meantime, Glister (1941), at Oxford University reported the produc-
tion of an antibacterial substance effective against gram-negative and
gram -positive bacteria by a different strain of A . flavus (found by the writers
to be wholly typical of the species). He noted the possibility of relationship
between the substance with which he was working and that earlier reported
by White.

Following the work of Jones, Rake, and Hamre (1943), McKee and Mac-
Phillamy (1943) succeeded in demonstrating the production of a second and
entirely different antibacterial substance. In certain chemical properties,
and in its action on bacteria, this was found to resemble penicillin very
closely, but actual identity was not proved. In a subsequent and more
detailed report, McKee, Rake, and Houck (1944) defined more exactly its
bactericidal action against various gram-negative and gram-positive bac-
teria and proved additional evidence of its penicillin-like characters. They
designated the substance i ''flavicidin'".

Concurrent with this work, Bush and Goth (1943a and 1943b), working
at Vanderbilt University, succeeded in demonstrating the production of an
antibacterial substance from still another strain of A. flavus which was
strongly active against Staphylococcus and other gram -positive forms but
comparatively inactive against gram-negative forms belonging to the E.
coli group. The substance was termed "flavicin". Identity with flavicidin
and with penicillin is possible but has not yet been proved. Cook and
Lacey (1944) report the production of appreciable amounts of an antibiotic
substance from a strain of A. parasiticus. This was provisionally desig-
nated "parasiticin", and its similarity to penicillin was noted. Identity
with flavicin (Bush and Goth) and flavicidin (McKee, Rake, and Houck)
was suggested. Since A. parasiticus is so closely related to A. flavus, it
would seem probable that an antibiotic produced by it would be similar to
that produced by the latter species under the same conditions.

Waksman and Bugie (1943) investigated a large number of strains be-
longing to the A. flavus-oryzae group. They found strains of A. oryzae to
show little activity, whereas strains of A. flavus showed increased but vary-
ing amounts. Yields' were markedly influenced by various nutritional and
environmental factors. Two types Of antibacterial substances were ob-
served: aspergillic acid and a substance similar to, if not identical with,
penicillin. When grown in submerged culture, one strain was found to
produce amounts comparable to the best strains of Penicillium notatum
tested. Unfortunately, yields were not quantitatively determined.

Chapter XXI
THE ASPERGILLUS OCHRACEUS GROUP

Outstanding Characters

Conidial heads ranging from sulphur yellow to varying shades of ochra-
ceous, depending upon the species and strain, showing a greenish tint
only in the single species A. sparsus; heads globose or radiate with
conidial chains commonly adhering into divergent columns.

Conidiophores normally showing shades of yellow in the outer layers of
the wall which is rough or pitted, usually prominently but occasionally
reduced to traces which are seen most readily in dry mounts.

Sterigmata in two series with the primary often quite large and septate.

Conidia in some series thin -walled and smooth, in others showing defi-
nitely double walls, more or less roughened or echinulate.

Sclerotia present in most species and strains, often dominating the cul-
tures; in others entirely lacking. When present, ranging in color from
cream or buff through pink and orange shades to purplish-vinaceous.

Molds belonging to this group are common wherever organic matter is
decomposing under natural conditions. In spite of great variation in super-
ficial appearance, length of conidiophore, size of heads, intensity and shades
of color, and sclerotium production, they fit together into a great natural
group of related forms. Extreme variants in the several series can be easily
considered separate species and have been so described, but collections of
great numbers of such forms present so many gradations that identification
by description becomes doubtful if not impossible. Allocation to series
centered upon some described species gives a practical method of grouping
together closely related members of the great aggregate.

The name "ochraceus", derived from the pigment ocher and attached to
the most abundant series in the group, is an old mycological usage which was
more or less definitely followed in Saccardo's use of it for a plaque in his
Chromotaxia. Ridgway analyzed the colors more exactly, dropped the
term ochraceus but included a plaque near to it as PI. XV, Col. 15 YO,
Ochraceous-Buff . The strains of this group, however, mostly show colors
closer to the yellower tints in Ridgway's Plate XXX, column 19. It is im-
portant that relationship with the great group shall be quickly grasped
whether exact identity with particular strains already known is claimed or
not.

Group characterization : The colony appearance of this group of Asper-
gilli varies greatly with the presence or absence of sclerotia. Some species

273

274 A MANUAL OF THE ASPERGILLI

are typically ochraceous in color from abundant conidial heads and are not
known to produce sclerotia. Others produce a few sclerotia, or clusters of
sclerotia, under very special conditions but still are characterized by their
abundant conidial heads. Others always produce sclerotia in sufficient
number to dominate the colony. Sclerotia, when present, are globose or
elliptical, 0.5 mm. or more in long axis, and vary in color from shades of
yellow to orange-vinaceous or even purplish. The submerged mycelium
varies from colorless to yellow, orange, or purplish shades. Heads are
globose or radiate and vary in color from bright yellow near citrine through
pale yellows to varying shades of ochraceous but never appear in umber
shades, and show green (near olivine, Ridgway, PI. XXXII) only in A.
sparsus. Conidiophores vary greatly in length and diameter but have
walls yellow, especially in the outer layer, pitted or rough — in occasional
dwarf strains with fairly thin walls the pitting becomes difficult to demon-
strate, hence careful examination with a good oil immersion objective may
be necessary. Vesicles are globose, or occasionally somewhat elliptical,
varying greatly in size with walls usually colorless and much thinner than
the conidiophore wall. Sterigmata are always in two series, with primary
sterigmata varying, as in the A. niger group, from fairly short to very long,
but commonly 15 to 30m upon Czapek's solution agar and occasionally much
longer on special substrata; secondary sterigmata differ less conspicuously
than primary sterigmata, being commonly 7 to 10/x by 1.5 to 2.5m; both
series of sterigmata are almost colorless when examined with high magnifi-
cations. Conidia are small, sometimes elliptical, but mostly globose or
subglobose, smooth in most strains, but in others very delicately wrinkled
or spinulose without prominent tubercles or bars of color, mostly 2 to 4/x in
long axis, larger in occasional strains.

The following key is an attempt to arrange this group into a satisfactory
order to bring nearly related organisms together. For this purpose specific
names already in the literature with the particular distinguishing marks cited
in their diagnoses are introduced as more or less definitely fixed points in
the group with which new material or unknown material can be compared.

Group Key

I. Conidial heads in fairly pure yellow tints such as sulphureus and citrinus

The A. sulphureus series

A. Sclerotia usually not abundant; conidiophores up to 1000m long; heads

abundant, radiate; vesicles globose; conidia globose or subglobose,
smooth, about 3m A. sulphureus Fresenius

B. Sclerotia abundant, commonly characterizing the culture; heads scattered,

or grouped in restricted areas, particularly at the drying margin of agar
slant cultures; conidia smooth, up to 4.0m

A. quercinus (Bainier) Thorn and Church
II. Conidial heads in pale to darker ochraceous shades. . . The A. ochraceus series

THE ASPERGILLUS OCHRACEUS GROUP 275

A. Conidia thin-walled, smooth A. ochraceus sub-series glaber

1. Sclerotia dominating the colony appearance; conidia small, about 3.0m-

a. Sclerotia abundant, buff to pink or purplish in color in age; heads

hemispherical to columnar, cream-buff to pale ochraceous

A . sclerotiorum Huber

b. Sclerotia abundant, at first gray, quickly becoming black; heads

globose or radiate; upon onions and other bulbs

A. alliaceus Thorn and Church
(A. uentii group, p. 241)

2. Sclerotia present but secondary to the crowded, honey-yellow conidial

heads; conidia smooth A. melleus Yukawa

B. Conidia with firmer walls, echinulate or barred

A. ochraceus sub-series echinulata

1. Conidia elliptical to subglobose.

a. Conidia 3.5 to 5.0m in long axis; echinulate heads ochraceous,

radiate, and splitting; sclerotia in type material

A. ochraceus Wilhelm

b. Conidia 3.0 to 3.5m, spinulose; sclerotia not found

A. elegans Gasperini

c. Conidia up to 6.3m in diameter, rough (Otherwise A . ochraceus)

S. buturacea Bainier

2. Conidia globose, echinulate, 7.0 to 8.0m in diameter (Otherwise A.

ochraceus) A. delacroixii (Sacc.) Thom and Church

3. Conidia strongly elliptical or pyriform, spinulose, 4.0 to 6.0m by 3.0 to

4.5M A. ostianus Wehmer

III. Conidial heads commonly showing a definite greenish tint, near olive-buff to
olivine in color A. sparsus Raper and Thom

Aspergillus sulphureus (Fres.) Thom and Church, in The Aspergilli, p. 185.

1926.

Syn.: S. sulphured Fresenius, in Beitr. zur Mykologie, Heft 3, p. 83, taf.
XI, fig. 30-33. 1863. See also Sacc. Sylloge 4: 73. 1886.

Colonies upon Czapek's solution agar at laboratory temperature, growing
rapidly, forming a variously wrinkled to zonate felt, white or tinged with
yellow to pink or purplish shades. Conidial heads irregularly produced in
most cultures, in pale or sulphur yellor tints, mostly globose, but often split-
ting into spore columns in age. Conidiophores with walls firm, rough or
pitted, yellow, varying greatly with the strain from very short and sparingly
produced to clustered in areas and dominating the culture, or entirely cover-
ing the mycelium where sclerotia are absent. Vesicles typically globose and
fertile over the whole surface. Radiating primary sterigmata closely
packed on the vesicular surface, varying from 8 to 10m by 3 to 5/x to several
times larger in some big heads, secondary sterigmata usually uniform,
phialiform about 8 to 10m by 2 to 3m- Conidia small, globose or subglobose
2 to 3 or 3.5m in long axis, thin-walled, smooth.

A . sulphureus as described by Fresenius had long conidiophores and yel-

276 A MANUAL OF THE ASPERGILLI

low globose heads. Sclerotia were not reported. In our experience, how-
ever, they are seen not infrequently in cultures which otherwise fit the
description of A. sulphureus as presented. In the sub-series, represented
by A. quercinus (Bainier, Thom and Church), sclerotia are very abundant
and heads comparatively few and scattered among the sclerotia. In addi-
tion to these two names which roughly designate sections or series of
isolates as we find them, every gradation between these extremes may be
anticipated.

The species is fairly common in soils, on cereal grains, and upon decaying
vegetation.

Several species have been described as having conidial heads sulphur
yellow but doubtfully separable from A. sulphureus.

S. ochroleuca Spegazzini, in Myc. Argent V, p. 434, in Anal. Mus. Nac. Buenos
Aires, Ser. 3, T. 13. 1911.

S. auricoma Gueguen, in Bui. Soc. Myc. France 16: 171-187, figs. 1-48. 1899. This
appears to have been a member of the series with primary sterigmata proliferating
abundantly to form little conidiophores and secondary heads thus giving the head the
appearance of bearing yellow hair. Such proliferation occasionally occurs in many
species but has not again been found in this group.

S. vitelline, Ridley, in Jour. Bot. (London) 34: 152, pi. 257, figs. 14-16. 1896, is
described as producing bundles or coremia composed of partially adherent conidio-
phores. The organism does not appear to have been cultivated and has not since
been reported.

Aspergillus quercinus (Bainier) Thom and Church, in The Aspergilli, p. 186,

1926.

Synonym: S. quercina Bainier, in Bull. Soc. Bot. France 28: 78. 1881.
See also Sartory, Etude biologique du Sterigmatocystis
quercina Bainier, in Bui. Soc. Myc. France 26: 349. 1910.

Colonies upon Czapek's solution agar spreading broadly, characterized
by the presence of an aerial white mycelium and the abundant production
of sclerotia over the whole area (PI. VII, E and fig. 74 A), or in sectors, or
variously distributed ; the mass changing from yellow to orange-yellow and
finally to rufous or brick red shades with the ripening of the sclerotia; re-
verse in shades of yellowish-orange. Conidial heads in yellow tints, near
sulphureus, scattered among the sclerotia or occurring in long-stalked
groups in the dryer areas of the culture tube or plate, mostly up to 200>
in diameter, but occasionally larger, up to 300 or even 400/x (fig. 74 C).
Conidiophores with walls mostly pale yellow, especially in the outer layer,
pitted (fig. 74 D), occasionally with abundant granules, about 2/jl in thick-
ness, varying from short and inconspicuous in crowded sclerotial areas to
very long tufts in dryer areas, up to 2 or even several millimeters by 10 to
20m- Vesicles colorless, crushing easily, 35 to 45^ in diameter, fertile over

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l**v#**otw

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Fig. 74. A-Z), Aspergillus quercinus, NRRL No. 394: A, Colonies on Czapek's
solution agar, 10 days, predominantly sclerotial but with localized areas of heavy
conidial production in central areas. B, Marginal area of the same colony showing
very abundant sclerotia, and scattered conidial heads in lower right-hand corner.
C, Conidial heads on hay infusion agar plate, X 18. D, Single head showing globose
character, crowded sterigmata, and roughened conidiophore. E, Aspergill
tiorum on Czapek's solution agar, 10 days. F, Portion of the same enlarge(
abundant large sclerotia and scattered comparatively small heads, X 6.

277

us sclero-
rged to show

278 A MANUAL OF THE ASPERGILLI

the entire surface. Sterigmata in two series: primary 10 to 20 or even 30m
in larger heads by 2 to 4 or 7 m at the tips; secondary 10 by 2 to
2.5/x. Conidia 2.5 to 3m by 3 to 3.5/* to almost globose, very nearly colorless
in mounts, smooth, thin-walled. Sclerotia white to brick red, up to 500m
in diameter (fig. 74 B).

The original culture as described by Bainier in 1880 was not preserved.
Neither was that of Sartory in 1910. However, cultures of organisms vary-
ing about as here described and complying fairly closely with Bainier's
description are not infrequently encountered from widely separated regions.

Aspergillus sachari Chaudhuri and Sachar, in Ann. Myc. 32: 95. 1934.

This organism was placed by the describers near ^4. quercinus on the basis of the
colors of the head and sclerotia. They ignored the colorless, smooth conidiophores
which, if correctly observed, would make their placing untenable. It is left here
arbitrarily until rediscovered and its characters verified. Compare: A. candidus
group.

Species characterization based upon the original description follows:

Colonies on Czapek's solution agar naphthalene yellow, (Ridgway, PI. XVI. 23,
yellow, f.) more or less floccose; reverse colorless at first then in yellow shades. Scle-
rotia scattered through the colony, quickly developed, hard, white through yellow
shades to cinnamon, up to 2 mm. in long axis with a depression in the center. Heads
abundant, radiate, 50 to 85m in diameter, or somewhat columnar in age up to 160 by
100m. Conidiophores 700 to 9C0m by about 8m with smooth colorless walls, about 1.5m
thick; vesicles globose, 20 to 30m in diameter; sterigmata in two series, covering the
whole vesicle, radiate; primary 5.4 by 2m, secondary 7.2 by 1.8m, conidia colorless,
smooth, globose, 2 to 2.7m in diameter. Found in alkaline soil.

Aspergillus sclerotiorum Huber, in Phytopathology 23 (3):
306-8, Fig. I. 1933.

Colonies on Czapek's solution agar rather slow growing, forming a smooth
mycelial layer irregularly overgrown with loose tufts of sterile hyphae within
which numerous sclerotia develop and often dominate the colony appear-
ance (fig. 74 E), white to shades of yellowish or ochraceous; reverse cream.
Conidial heads given as sulphur yellow by Huber, but in our cultures found
in ochraceous shades such as cream-buff (Ridgway, PI. XXX), mostly
appearing as columnar masses of conidia up to 250m long, sometimes
splitting, 60 to 70m at the base and up to 125m at the apex. Conidiophores
with walls yellow and pitted, commonly 200 to 400m long by 7 to 10m in
diameter, but varying from very short, 50m in length, to occasional groups
up to 1200m; in old cultures, arising as branches from loose aerial hyphae.
Vesicles subglobose, 15 to 20m in long axis. Sterigmata in two series, pri-
mary usually 8 to 9m, occasionally much more but not above 20m long by 3.5
to 4.5m in diameter, secondary 8 to 9m by 2m- Conidia smooth, mostly 2
to 2.5m- Sclerotia abundant (fig. 74 F), beginning to appear within 3 days,
white to cream or pink, up to 1.5 mm. in diameter, well scattered over the

THE ASPERGILLUS OCHRACEUS GROUP 279

mycelium or vaguely in zones, giving the characteristic appearance to the
colony, hence the name.

The type culture NRRL Xo. 415 (Thorn No. 5351 received from Huber)
was isolated from rotting apples in Oregon and proved capable of causing
decay in apples under controlled conditions in which other strains failed to
produce rot. It has not been discussed by others. Blochwitz (Ann.
Mycol. 32(1/2) : 88. 1934) considered A. sclerotiorum to be a synonym for
A. clegans Gasperini, but this is not consistent with the description.

Aspergillus melleus Yukawa, in Jour. Coll. Agr. Imp. Univ.
Tokyo 1, No. 3, p. 366, Taf. 16. 1911.

Colonies on Czapek's solution agar rapidly growing and spreading, form-
ing an aerial felt of sterile hyphae and conidiophores, and producing
sclerotia with walls white to yellowish-brown, commonly a shade of yellow-
orange near "melleus" of Saccardo's Chromotaxia (Approximately Ridgway
PI. XXX, 19", honey yellow; or warm buff, PI. XV, 17'd); reverse reddish
to brown shades. Conidial heads ranging from small to large and globose,
commonly splitting into dense columnar masses, sometimes 250 to 300m
in long axis. Conidiophores with walls yellow, pitted, up to 500 or even
1000/x long, 15 to 20 or even 25m in diameter. Vesicles up to 40 to 50m in
diameter, globose in large heads, more or less pyriform in small heads,
fragile and readily crushed in mounting. Sterigmata in two series, primary
10 to 20/x by 2.5 to 4/x, secondary commonly 3 to 10m by 2 to 3/x, with oc-
casional larger sterigmata in either series. Conidia in long chains, almost
colorless when mounted, very thin-walled, smooth, slightly elliptical, about
3m or a little more in long axis. Sclerotia ranging from 400 to 700m in
diameter, in yellow-brown shades.

Hanzawa furnished Thorn's culture No. 4291.6 (NRRL No. 416) as the
type strain used by Yukawa. Other strains, including cultures from For-
mosa, South Africa, and the United States, indicate that the cultural aspect
of Yukawa's species is carried by organisms widely distributed. Some re-
lated strain may have been described by Tiraboschi as A. ochraceus var.
microspora (in Ann. di Botanica VII. 14. 1908) from corn bread in Italy.
This name is used also by Nakazawa (Jour. Agr. Chem. Soc. Japan 10:
English summary p. 185. 1934).

Aspergillus ochraceus Wilhelm, in Inaug. Diss. Strassburg, p. 66. 1877.
Wilhelm's exsiccati exist as Rabh. Fl. Europaei no. 2361.

Colonies upon Czapek's solution agar spreading fairly broadly, usually
plane, zonate in some strains, characterized by a tough submerged felt which
may be colorless, yellow, orange, or purplish and, arising from this, more or
less crowded conidial structures which give to the colony its characteristic

280

A MANUAL OF THE ASPERGILLI

color and appearance (PI. VII F and fig. 75 A) ; reverse ranges from colorless
through orange to purplish shades.

Conidial heads when large mostly globose or variously splitting into
masses of conidial chains (fig. 75 C), variously colored from very pale to
deep ochraceous shades . Conidiophores vary greatly in length and diameter

Fig. 75. Aspergillus ochraceus series. A and B, A. ochraceus, NRRL No. 398
and No. 408, respectively, growing on Czapek's solution agar at room temperature,
10 days. C, Mature heads of strain No. 398; the tendency to split into divergent
columns in age is characteristic. D, Photomicrograph of a single head showing globose
vesicle, sterigmata in two series, and coarsely roughened conidiophore, X 325.

in the various strains, but typically show a yellow color in the outer layers
of the thick wall which shows characteristic pitting or roughening (fig.
75 D). Vesicles mostly globose or somewhat elliptical, and fertile over the
whole surface (fig. 75 D). Sterigmata in two series: primary varying from
small to very large, commonly 15 to 30/x in length; secondary fairly uniform,

THE ASPERGILLUS OCHRACEUS GROUP 281

commonly 7 to 10/x by 1.5 to 2.5/x. Conidia globose, subglobose, or some-
what elliptical, more or less rough or echinulate, ranging from 3.5 to 5.0m
in long axis. No perithecia reported. Sclerotia are present in Wilhelm's
material preserved in exsiccati.

Among the great number of isolates belonging to this group a considerable
number show approximately the characters as drawn from Wilhelm's
description and his exsiccati. In the limited sense then, ^4. ochraceus
Wilhelm can be interpreted to apply to those ochraceous strains which bear
echinulate conidia from 3.5 to 5.0m in diameter and produce sclerotia.
Among the strains included, almost all are found to produce colonies of
consistent aspect in continuous culture.

Aspergillus elegans Gasperini, in Atti. Soc. Toscana Sci. Nat. Pisa

Mem., 8: 328, fasc. 2. 1887.

Synonym: S. elegans (Gasp.) Sacc, in Syll. 10: 525.

This species by description differs little from A. ochraceus Wilhelm
except for the absence of sclerotia, and conidia which do not exceed 3.5/x
in diameter. Thorn and Church did not recognize it as a valid species in
1926, and one may be certain that no sharp line can be drawn separating
A. elegans from A. ochraceus. Nevertheless, forms producing conidia con-
sistently less than 3. 5m are commonly encountered among miscellaneous
isolations from nature, and there is an argument for retaining a species to
include such forms. The following species diagnosis, taken from Saccardo
(10: 525) was presented by Thorn and Church (1926):

"Mycelium white; stalks continuous, unbranched, hyaline then pale
ochraceous, 1 to 6 mm. long, by 5 to 12/x in diameter, delicately studded with
drops ; vesicle up to 70m diameter, radiate, entirely covered with sterigmata;
sterigmata, primary 4 to 26m long, secondary 7 to 14m long by 1 to 2m;
conidia ochraceous, elliptical to globose, up to 3 to 3.5m, with wall very deli-
cately verruculose, sclerotia not found."

Various other species have been described which are obviously closely
related and belong to the A. ochraceus series. A partial list would in-
clude :

S. helva Bainier, in Bull. Soc. Bot. France 28: 78. 1881. Thorn's culture No.
4640.476, received under this name, came from the Bainier collection.

A. alutaceus Berkeley and Curtis (in Grevillea 3, No. 25, p. 108. 1875) was the
name proposed for a mold found upon corn. The specimen is preserved as No. 3793
in Curtis' Herbarium now in the Cryptogamic Herbarium of Harvard University.
The specimen shows that this species was probably a strain of A. ochraceus.

A. ochraceus var. microspora Tiraboschi (in Ann. di Bot. [Rome] 7: 14. 1908)
represents a strain in which all measurements were reported as reduced.

A. rehmii Zukal. A culture from Dr. Westerdijk, received under this name, is also
a member of this series.

282 A MANUAL OF THE ASPERGILLI

Certain large-spored species of doubtful validity and relationship have
been described that are believed to represent members of the A. ochraceus
series. Although obviously rare, these species were described with suf-
ficiently distinctive characters to warrant their retention. With continued
search, it is possible that they may be reisolated.

Aspergillus delacroixii (Sacc.) Thorn and Church, in The
Aspergilli, p. 190. 192G.

Synonym: S. ochracea Delacroix, in Bull. Soc. Myc. France 7: 109,
PI. VII, fig. f. 1891. (Delacroix failed to recognize the
previous use of the specific name.)
S. delacroixii Sacc, in Sacc. 10: 527.

This species is reported as having conidiophores pale yellow and rough,
500 to 1000m in length; vesicle globose, thick-walled, punctate, yellow;
sterigmata in two series, primary 39/x by 12m, secondary (from Delacroix's
figures) about 8 to 10m by 2 to 3m; conidia globose, finely roughened, 7 to 8m
in diameter. The yellow and roughened conidiophores ally this species with
the A. ochraceus groups hence would justify the description, if a form with
such large conidia should be found again. Otherwise, it is possible that
some old material of a strain of A. oryzae might have furnished the type.

Aspergillus butyracea (Bainier) n. comb.

Synonym: S. butyracea Bainier, in Bull. Soc. Bot. France 27: 29. 1880.

Specimen attributed to Bainier in C. Roumeguere's Fungi

Gallici Exsiccati No. 995.

This is described as a large-spored strain belonging in this group. The
specimen showed a black Aspergillus, as well as an ochraceous form with
spores up to about 6m and rough. Colonies butter yellow, including
conidiophores, heads, and conidia; conidiophores yellow, in mounts finely
punctate or pitted, 13 to 16m in diameter; sterigmata primary up to 25m,
secondary 10 to 12m in length; conidia described as smooth 5.2m, but those
found in the material were rough and up to 6.3m- It is entirely possible
that this species may be found again, but is has not been reported since
Bainier described it.

A. penicillopsis (Henngs.) Racib., P. Hennings Synonym Stilbothamnium
penicUlopsis P. Henn. and E. Nym. described in Fungi Monsunenses
(Warburg. O. Monsunia, Bd. I, p. 37. 1900. Leipzig); exsiccati of
type in Pathological Collections, U. S. Dept. Agr. Bureau of Plant In-
dustry, as: Raciborski no. 87, in Crypt. Parasiticae Java.

This material shows an Aspergillus with the color and general appearance
of .4. ochraceus but of gigantic proportions. Measurements as follows:

THE ASPERGILLUS OCHRACEUS GROUP 283

Conidiophore 50 to 70m in diameter, 10 mm. or more long with walls 7 to
12/x thick. Surface ragged and more or less pitted. Vesicle up to 175m
in diameter with walls 7m thick, marked with a deep pit for each sterigma.
Sterigmata, primary 50 to 90m, at times 120 by 8 to 10m at the outer end,
sometimes with one cross wall; secondary 15 to 25m by 3 to 4m, clustered on
the apex of the primary. Occasionally with a sterile cell interposed between
secondaries and primaries. Conidia 8 to 12m by 5 to 8m, elliptical, pitted,
yellowish.

Gigantic forms such as this occasionally appear under field conditions,
hence reach fungus herbaria. Whether these are really different from some
of the usual species can only be determined by collection and laboratory
cultivation. Until such study has been made, such forms as A. penicil-
lopsis must be questioned.

Aspergillus ostianus Wehmer, in Bot. Centralb. 80: 449-461. 1899; also
Monogr. pps. 117-119, Taf. II, No. 1. 1899-1901.

Colonies upon Czapek's solution agar growing fairly well, producing a
surface growth of crowded conidiophores and conidial heads in yellowish
to ochraceous shades, passing to shades of cinnamon in very old cultures
with reddish-brown colors in reverse (Wehmer reported rusty-yellow, pale
to deep brownish-yellow to cinnamon). Conidial heads globose, up to
200m in diameter. Conidiophores yellow, coarsely roughened, mostly 500
to 700m long in crowded areas, becoming 1 to 2 mm. in margins of old
colonies and usually 7 to 10m in diameter, with walls heavy, up to 2 or 2.5m
in thickness. Vesicles commonly globose, about 40m m diameter, oc-
casionally much larger up to 70m, hi growing heads thin-walled, colorless,
crushing easily, leaving the funnel-like yellow tip of the conidiophore open.
Sterigmata in two series: primary from 15 to 20m long in smaller heads
to 35m by about 8m at the tip in large heads, secondary 10 to 13m by about
3m- Conidia commonly 3 to 4m or even 5m in long axis, varying from pyri-
forrrj to elliptical, or at times subglobose, rough. Sclerotia occasionally
present but not conspicuous.

This diagnosis was based upon Wehmer 's description, and is reasonably
well represented by strain NRRL No. 420 (Thorn No. 4724.35) received
from Raistrick in 1924 and reported by him to have come from Westerdijk
as Wehmer 's original strain. With conspicuous appearances suggesting
relationship to the A. ochraceus group, the shape and ultimate color of the
ripe spores suggest a border line position between A. ochraceus and A.
tamarii.

Aspergillus sparsus Raper and Thorn, in Mycologia 36: 572-574,

fig. 6. 1944.

Colonies upon Czapek's solution agar at room temperature spreading
broadly, dull grayish-brown in color, at first largely submerged, but later

284

A MANUAL OF THE ASPERGILLI

developing limited aerial growth, giving rise to widely scattered erect
conidial structures (fig. 76 A) characterized by dull greenish-tan heads not
affecting the color of the colony as a whole; reverse in brown shades; odor
Colonies upon hay infusion agar spreading broadly, almost wholly

none.

submerged, giving rise to scattered but conspicuous conidial structures
(fig. 76 B), often in definite concentric zones; heads globose, radiate, in olive-

Fig. 76. Aspergillus sparsus: A and B, Colonies on malt extract and hay infusion
agars, respectively, 2 weeks; note scattered heads in B and almost complete absence
in A. C, Conidial heads, X 18. D, Single conidial head showing globose vesicle and
roughened conidiophore, X 425.

buff shades (Ridgway, PI. XL). Colonies upon malt extract agar spreading
irregularly, floccose, 1 to 2 mm. deep, cream-buff in color ; conidial structures
very few in number; heads globose, radiate, dull olive-yellow in color
(Ridgway, PI. XL). Conidial structures generally few in number, never
abundant, erect, arising from a submerged mycelium; heads typically glo-
bose becoming more or less radiate in age (fig. 76 C), mostly 200 to 250m

THE ASPERGILLUS OCHRACEUS GROUP 285

in diameter, occasionally as much as 500m, pale olive-buff to olive-buff
(Ridgway, PI. XL) upon Czapek and hay infusion agars to pale olivine or
olivine (Ridgway, PI. XXXII) upon malt extract agar. Conidiophores
straight, mostly 1 to \\ mm. in length by 10 to 12m in diameter, approxi-
mately uniform in diameter throughout, wall 1.2 to 1.5m in thickness,
conspicuously echinulate, typically arising from a foot cell enmeshed in a
network of "feeder" hyphae, often tapering abruptly in the region im-
mediately beneath the vesicle. Vesicle comparatively thin-walled, globose
(fig. 76 D), mostly 40 to 50m in diameter, occasionally larger or smaller,
bearing sterigmata over the entire surface. Sterigmata in two series,
primaries crowded, comparatively short and stout, commonly 8 to 10m by
3 to 5m, secondaries 6 to 8m by 2.5 to 3.5m- Conidia pale yellowish in mass,
individually showing slight coloration, subglobose to slightly elliptical,
very finely roughened, mostly 3 to 3.5m in long axis.

Type culture NRRL No. 1933 was isolated in February 1943, from soil
collected in La Lima, Honduras, by Dr. L. A. Llnderkofler. A second strain
which duplicates the type almost exactly was subsequently isolated from
soil collected in Bixar County, Texas, by Sister Mary Clare of Our Lady of
the Lake College, San Antonio, Texas.

The correct position of this species within the genus Aspergillus is open to
question. The presence of a colored, coarsely roughened conidiophore indi-
cates close relationship with Aspergillus ochraceas. This is likewise sup-
ported by the globose vesicle and head, although these characters are typical
of other groups as well. The scarcity of fruiting structures upon all media,
and more particularly the greenish tint of the spore masses, however, tend
to set it apart from the common representatives of this great group. The
general habit of the colonies together with the paucity of conidial structures
is strongly suggestive of Aspergillus alliaceus, but this latter species does not
show any trace of greenish color in its conidial heads; it does possess smooth,
colorless conidiophores, and upon ordinary culture media regularly produces
an abundance of black sclerotia which very often dominates and charac-
terizes the culture. In the color and character of its conidial heads A.
sparsus is somewhat suggestive of George Smith's new species, A. avenaceus
(Trans. Bui. Mycol. Soc. 25: 24-27, PI. I. 1943), but it differs from this,
as it does from A. alliaceus, in possessing rough conidiophores and in its
failure to produce sclerotia. Until additional related forms are isolated,
we believe it best to consider this species as a member of the A . ochraceus
group, realizing that it does not entirely fit this placement as the group has
hitherto been considered.

Occurrence and Economic Importance

Members of the A . ochraceus group are widely distributed in nature and
can be obtained from a variety of sources. They are especially common

286 A MANUAL OF THE ASPERGILLI

in soils and have been isolated from samples collected in many parts of the
world. Within the group, strains approximating the species A. ochraceus
are by far the most abundant, although heavy sclerotium-producing strains
approximating A. quercinus are not infrequently encountered. They are a
common component of the microflora of decaying vegetation, but there is
little evidence that they play a very active role in processes of decomposi-
tion.

Huber (1933) found a member of the group, A. sclerotiorum, capable of
rotting apples and pears. Members of the group are commonly found in
musty or moldy cereal grains, but are not as characteristic of this sub-
stratum as is Aspergillus candidus and members of the A. glaucus group.
In at least one instance A. ochraceus was reported as a human pathogen
(Ceni, 1905).

In the Orient, A. ochraceus and allied species constitute a portion of the
mold flora characteristically found on "Katsuobushi" and other fermented
preparations made from fish (Yukawa, 191 1). Aspergillus melleus Yukawa
was isolated from such material. Because of the mixture of forms present,
including members of the A. glaucus and A . flavus-oryzae groups, it is prob-
ably incorrect to say that any particular species or group of species is
responsible for this fermentation. (See also Hanzawa, 1911).

A. ochraceus has been used to bring about desired changes in the flavor
of coffee, and its use is covered by U. S. Patent No. 1,313,209. Samples
of the fermenting coffee showed the organism used to be a strain of A.
ochraceus indistinguishable from Wilhelm's species. Whereas A. niger,
A . tamarii, and A . flavus were also capable of developing in the fermenting
coffee, A. ochraceus alone of the species tried gave a satisfactory flavor.

As a whole, the A. ochraceus group constitutes a very abundant, but little
studied, group of molds. Whether the scarcity of published reports re-
garding biochemical activities indicates an absence of such, or whether it
merely reflects a limited amount of investigation, can at present only be
guessed.

PART III

REFERENCE MATERIAL

Chapter XXII
TOPICAL BIBLIOGRAPHY

In preparing this manual, the writers have considered it inadvisable to
attempt to discuss the biochemical activities of the Aspergilli, since this
subject is book-length in itself. Furthermore, the aim of the manual is to
provide the mycologist and microbiologist with a means of identifying and
interpreting Aspergilli as they are isolated from nature and to furnish the
chemist, working with molds, with a guide which will enable him to main-
tain industrially important cultures in an optimum condition. Neverthe-
less, because of the increasing importance of the Aspergilli as agents re-
sponsible for industrial fermentations, as subjects for physiological and
biochemical investigations, and now as possible sources of various anti-
biotics, it has seemed advisable to present a topical bibliography dealing
with these and related subjects. We have not attempted to present a
complete bibliography, but rather to present a sufficient number of refer-
ences to provide the investigator and student with an entree to the literature
of particular fields. An attempt has been made to choose the more im-
portant papers for citation. Believing that more recent contributions will
generally be of the greatest interest and value to the user, this topical
bibliography is presented in chronological, rather than alphabetical, order.
A list of subjects under which references are presented follows.

CONTENTS OF TOPICAL BIBLIOGRAPHY

Acid Production by Aspergilli 290

Citric Acid 290

Fumaric Acid 293

Gallic Acid and Tannin Fermentation 294

General Papers and Reviews 294

Gluconic Acid 295

Itaconic Acid 297

Kojic Acid 297

Oxalic Acid 298

Miscellaneous Acids 299

Antibiotics and Toxins 300

Chemistry of Mold Tissue 301

Enzyme Production by Aspergilli 302

Aspergillus flavus-oryzae group 302

Aspergillus niger group 304

Aspergilli, General 305

Fat Production by Aspergilli 306

Pathogenicity of the Aspergilli 307

Physiology of the Aspergilli 310

289

290 A MANUAL OF THE ASPERGILLI

Pigments and Coloring Substances 312

Soil Tests for Mineral Deficiencies 313

Variation in the Aspergilli 314

Vitamins and Growth Substances 316

Miscellaneous Products 316

Alcohol 316

Chitin 317

Ergosterol 317

Fluorescein 317

Gums 317

Hydroxylamine 317

Mannitol 317

Polysaccharides 31S

Ochracin 318

Terrein 318

Acid Production

Citric Acid

Wehmer, C. 1893. Note sur la fermentation citrique. Bull. Soc. Chim. France 9:

728-730.
Wehmer, C. 1893. Preparation d'acide citrique de synthese, par la fermentation

du glucose. Compt. Rend. 117: 332.
Wehmer, C. 1894. Process of making citric acid. U. S. Patent 515,033. Febru-
ary 20.
Wehmer, C. 1897. Uber Zwei weitere freie Citronensaure bildende Pilze. Chem.

Ztg. 21: 1022-1023.
Wehmer, C. 1912. Uber Citronsauergarung. Chem. Ztg. 36: 1106-1107.
Currie, J. N. 1917. The citric acid fermentation of Aspergillus niger. Jour. Biol.

Chem. 31: 15-37, pis. 2, figs. 5.
Molliard, M. 1919. Production d'acide citrique par le Sterigmatocystis nigra.

Compt. Rend. Acad. Sci. (Paris) 168: 360-363.
Butkewitsch, W. 1923. Uber die "Citronensauregarung." Biochem. Ztschr. 142:

195-211.
Molliard, M. 1924. Influence de la nature des sucres sur la formation d'acides

organiques par le Sterigmatocystis nigra en milieu desequilibre. Compt.

Rend. Soc. Biol. (Paris) 90: 1395-1397.
Wehmer, C. 1925. Bildung von citronensaure aus glykonsaure durch pilze. Ber.

Deut. Chem. Gesell. 58: 2616-2619.
Bernhauer, K. 1926. liber die Saurebildung durch Aspergillus niger. III. Die

Bedingungen der Zitronensaurebildung. Biochem. Zeitschr. 172: 324-49.
Henneberg, W. 1926. Handbuch der garungsbakteriologie, 2 v., illus. Berlin.
Amelung, H. 1927. Beitrage zur Saurebildung durch Aspergillus niger. Zeit.

Physiol. Chem. 166: 161.
Challenger, F., Subramaniam, V. and Walker, T. K. 1927. The mechanism of

the formation of citric and oxalic acids from sugar by Aspergillus 'niger.

Jour. Chem. Soc. (London) 200-208.
Kostytschev, S. and Tchesnokov, W. 1927. Bildung von Zitronensaure und

Oxalsaure durch Aspergillus niger. Planta, Abt. E. Zeits. Wiss. Biol. 4:

181-200.

TOPICAL BIBLIOGRAPHY 291

Walker, Th. K., Subramaniam, V. and Challenger, F. 1927. The mechanism of
the formation of citric and oxalic acids from sugars by Aspergillus niger.
II. Jour. Chem. Soc, London, 3044-54.

Bernhauer, K. 1928. Zum Chemismus der Citronensaurebildung durch Pilze.

I. Die Saurebildung aus verschiedenen Kohlenstoffverbindungen. Biochem.
Zeitschr. 197: 309-26.

Bernhauer, K. 1928. Zum Chemismus der Citronensaurebildung durch Pilze.

II. Die Citronsaurebildung aus Gluconsaure. Biochem. Zeitschr. 197:
327-42.

Bernhauer, K. and Schon, K. 1928. Zum Chemismus der Citronensaurebildung
Durch Pilze. III. Mitteilung: Uber die Hypothesen Citronensaurebildung
und das Auftreten von Acetaldehyd in den Pilzkulturen. Biochem. Ztschr.
202: 164-179.

Bernhauer, K. 1928. Beitrage zur Enzymachemie der durch Aspergillus niger
Bewirkten Saurebildungsvorgange, I. Ztschr. Physiol. Chem. 177: 86-106.

Bernhauer, K. 1928. Uber die Saurebildung bei Aspergillus niger. IV. Die
Bedeutung der Myzelentwicklung fur die Saurebildung. Biochem. Zeitschr.
197: 287-308.

Fernbach, A. and»Yuill, J. L. 1928. Improvement in and relating to the processes
for the production of citric acid. U. S. Pat. 1,691,965 and 1,691,966.

Filosfov, M. S. and Malinovskii, V. E. 1928. O limonno-kislom drozenii.
Nauchnuie Zapiski 5: 235-239.

Szucs, J. 1928. Process for producing citric acid by means of fermentation.
Canad. Pat. 251,180. 1925, and U. S. Pat. 1,679,186.

Cahn, F. J. 1929. Perfectionnements a la fabrication de l'acide citrique par
fermentation. Fr. Pat . 675,236 and 675,237.

Challenger, F. 1929. The production of citric acid by fermentation process.
Ind. Chem. 5: 181-184.

Amelung, H. 1930. Wachstum und Saurebildung von Aspergillus niger unter
Wasser. Chem. Ztg. 54: 118.

Chrzaszcz, T. and Tiukow, D. 1930. Biochemische Umbildungen der Essigsaure
durch Schimmelpilze und liber den Chemismus der Citronensaurebildung.
Biochem. Zeit. 229: 343.

Bernhauer, K., SiEBENauGER, H. and Tschinkel, H. 1931. Zum Chemismus der
Citronensaurebildung durch Pilze. IV. tlber die Umwandlung der Zuck-
ersaure. Biochem. Ztschr. 230: 466-474.

Bernhauer, K. and SiEBENauGER, H. 1931. Zum Chemismus der Citronensaure-
bildung durch Pilze. V. Die Citronensaurebildung aus Essigsaure. Bio-
chem. Zeit. 240: 232-44.

Taylor, J. N. and Boyd, R. G. 1931. United States citric acid industry now self-
sufficient. U. S. Dept. Com., Bur. Foreign and Dom. Com., Com. Rpt. 4:
227-228.

Bernhauer, K. 1932. Die oxydative Garungen. Verlag Julius Springer, Berlin.

May, O. E. and Herrick, H. T. 1932. Production of organic acids from carbo-
hydrates by fermentation. U. S. Dept. Agr. Circular 216.

Porges, N. 1932. Citric Acid Production by Aspergillus niger. Am. Jour. Bot. 19:
559-567.

Sakaguchi, K. and Yamaya, J. 1932. Production of citric acid by molds. III.
Japanese black molds. Jour. Agr. Chem. Soc. Japan 8: 489-97.

Kostytschev, S. and Berg, V. 1933. Conditions of Biochemical Production of
Citric Acid. Bull. State Inst. Agr. Microbiol. USSR 5: 8-27.

292 A MANUAL OF THE ASPERGILLI

Butkewitsch, W. S., Menzschinskaja, E. W. and Trofimova, E. I. 1934. Zur
Biochemischen Herkunft von Citronen-und Oxalsaure. I. Uber die Bildung
von Citronensaure aus Essigsaure. Biochem. Zeit. 272: 290. II. Die
Mycelsubstanzen als ein Quelle der Saurebildung. Biochem. Zeit. 272: 364.

Doelger, W. P. and Prescott, S. C. 1934. Citric Acid Fermentation. Ind. Eng.
Chem. 26: 1142-49.

Sotnikov, E. 1934. Production of citric acid by Aspergillus niger. I. Production
of citric acid in unchanged solutions. Compt. Rend. Acad. Sci. USSR 3(4):

273.
Sotnikov, E. 1934. Production of citric acid by A spergillus niger. II. Production
of citric acid in changed solutions. Compt. Rend. Acad. Sci. USSR 3(4):

279-83.
Sotnikov, E. 1934. Production of citric acid by Aspergillus niger. III. Produc-
tion of citric acid in often-changed solutions. Compt. Rend. Acad. Sci.

USSR 3(7): 544-7.
Zubkova-Gitler, S. R. and Zalts, R. M. 1934. Preparation of Citric Acid from

Wood-Pulp Hydrolysates. Lesokhimicheskaya Prom. 3: 4-7.
Butkewitsch, V. S. and Gaewskaya, M. S. 1935. Yield of citric acid from sugar

as a basis for estimating the mechanism of its formation from the latter.

Compt. Rend. Acad. Sci. U.S.S.R. (N.S.) 3: 405-408. Abstracted in Chem.

Abstr.
Cahn, F. J. 1935. Citric acid fermentation on solid materials. Ind. Eng. and

Chem. 27: 201-204.

Chrzaszcz, T. and Peyros, E. 1935. Optimale Bedingungen der Citronensaurean-
haufung, sowie einiger Beobachtung zu Theorie der Citronsaurebildung.
Biochem. Ztschr. 280: 325-336.

Giordani, M. 1935. Fermentazione citrica. La Chimica e 1'Industria 17: 77-81.

Gudlet, M. A. 1935. Gasregime of Aspergillus niger as related to citric acid forma-
tion. Proc. Inst. Sci. Res. Food Ind. (Leningrad) 3, No. 1: 45-69. Ab-
stracted in Chem. Abst.

Ivanov, N. N. 1935. Biochemical production of citric acid. Proc. Inst. Sci. Re-
'search Ind. (Leningrad) 3, No. 1, 3-4. Practical and theoretical studies.

Kresling, E. K. 1935. Effect of radium on citric acid formation by A. niger.
Proc. Inst. Sci. Res. Food Ind. (Leningrad) 3(4/5): 130-145. No marked

effect.
Bernhauer, K. and Iglauer, A. 1936. Uber die Saurebildung aus Zucker durch

Aspergillus niger. VI. Faktoren der Citronensaureanhaufung. Biochem.

Ztschr. 286: 45-59.
Cahn, F. J. 1936. Citric Acid Manufacture. U. S. Patent 2,047,669. July 14.
Chrzaszcz, T. and M. Zakomorny. 1936. Uber die Bedeutung der Apfelsaure bei

der Umbildung der Essigsaure in Citronensaure durch verschiedene Schim-

melpilze. Biochem. Zeit. 285: 348-355.
Clutterbuck, P. W. 1936. Recent developments in the biochemistry of moulds.

Jour. Soc. Chem. Ind. 14: 55T-61T.
Ivanov, N. N. 1936. Biochemical production of citric acid. Proc. Inst. Sci. Re-
'search Ind. (Leningrad) 3, No. 4, 5. Laboratory and semiplant stages to

commercial production.
Lamb, A. R. 1936. Utilization of (Cane) Molasses. Proc. Hawaiian Sugar Planters'

Assn. 56th meeting, 42-3.
Wells, P. A., Moyer, A. J. and May, O. E. 1936. The chemistry of the citric acid

fermentation. I. The carbon balance. Jour. Amer. Chem. Soc. 55:555-558.

TOPICAL BIBLIOGRAPHY 293

Nakazawa, R., Takeda, Y and Nakano, M. 1937. Citric Acid Manufacture by
Fermentation. I. Aspergillus awamori var. fumeus. Jour. Agr. Chem.
Soc. Japan 13: 52-62.

L'vov, S. and Toupizina, G. M. 1938. Effect of sodium fluoride on the formation
of citric and gluconic acids by the fungus Aspergillus niger. Compt. Rend.
Acad. Sci. (USSR) 21: 307-11.

Wells, P. A. and Herrick, H. T. 1938. Citric Acid Industry. Ind. and Eng.
Chem. 30: 255-262.

Zender, J. 1938. Production of Citric Acid. U. S. Patent 2,121,063. June 21.

Bernhauer, K. 1939. Garungschemisches Praktikum. Zweite Ouflage. pp. 317.
Verlag Julius Springer, Berlin.

Zhuravskii, G. I. 1939. The Gas Exchange in Aspergillus niger during the Forma-
tion of Citric Acid. Microbiology 8: 414-430. (USSR)

Berhauer, K., Knobloch, H. and Zippelius, O. 1940. Uber die Saurebildung
aus Zucker durch Aspergillus niger. VIII. Der Einfluss von Magnesium
auf die Saurebildung. Biochem. Ztschr. 303: 300-307.

Das Gupta, G. C, Saha, K. C. and Guha, B. C. 1940. The fermentative produc-
tion of citric acid and oxalic acid from molasses. J. Indian Chem. Soc,
Ind. and News Ed. 3: 64-74.

Mel'nikova, A. A. and Trofimova, E. I. 1940. Activation of the process of citric
acid accumulation by surface molds of Aspergillus niger. Microbiology
(USSR) 9: 558-68.

Prescott, S. C. and Dunn, C. D. 1940. Industrial Microbiology. Chapt. XXV:
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Wang,Y. 1940. Citric acid metabolism of A spergillus carbonari us. Jour. Shanghai
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Barinova, S. A. 1941. Considerations in the appraisal of the circulating-medium
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Bernhauer, K., Iglauer, A., and Knobloch, H. 1941. Uber die Saurebildung
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Karow, Edward Otto. 1942. The Production of Citric Acid in Submerged Cul-
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Sztics, Joseph. 1944. Method of producing citric acid by fermentation. U. S.
Patent 2,353,771, July 18.

Fumaric Acid

Ehrlich, F. 1911. Uber die Bildung von Fumarsaure durch Schimmelpilze. Ber.
Deut. Chem. Ges. 44: 3737-3742.

Wehmer, C. 1918. Uber Fumarsaure-Garung des Zuckers. Ber. Deut. Chem.
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Ehrlich, F. 1919. Uber Fumarsaure-Garung des Zuckers. Ber. Deut. Chem. Ges.
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Schreyer, R. 1928. Sauerungsversuche mit dem Pilz Aspergillus fumaricus.
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Thies, W. 1930. Untersuchungen liber den einfluss der bedingungen auf die saure-
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294 A MANUAL OF THE ASPERGILLI

Thies, W. 1931. Decomposition of salts of organic acids by the mold fungus

Aspergillus fumaricus. Ber. 64B: 214-8.
Chrzaszcz, T. and Zakoronorny, M. 1933. Biochemical Transformation of Sugar

by Fungi. The Transformation of Fumaric Acid, the Accumulation of

Formic Acid, and the Chemistry of Oxalic Acid Formation. Biochem. Zeit.

259: 156.

Gallic Acid and Tannin Fermentation

Scheele, K. 1787. tTber das wesentliche gallapfelsalz. Crell's Chem. Ann. 1:

3-12.
Robiquet, E. 1852. Recherches sur la fermentation gallique. Jour. Pharm. 22:

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van Tieghem, Ph. 1867. Sur le fermentation gallique. Compt. Rend. Acad. Sci.

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Pottevin, H. 1900. La tannase, diastase dedoublant l'acide gallotannique.

Compt. Rend. Acad. Sci. Paris 131: 1215-1217.
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Biochem. Jour. 25: 752-5.
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Chi-Laing Kuo. 1939. Researches on Fermentation for the Production of Gallic

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Kwang-Chur Hsieh. 1939. Researches on Fermentation for the Production of

Gallic Acid. IV. Optimum concentration of gallnut extract and rate of

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Acid. V. Changes in tannin content, gallic acid content and total acidity

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General — Acid Production

Elfving, F. 1918-19. tJber die Bildung organischer Sauren durch Aspergillus
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Gluconic Acid

Molliard, M. 1922. Sur une nouvelle fermentation acide produite par le Sterig-

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296 A MANUAL OF THE ASPERGILLI

Schreyer, R. 1931. Comparative studies on the formation of gluconic acid by

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Moyer, A. J., Wells, P. A., Stubbs, J. J., Herrick, H. T. and May, O. E. 1937.
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Wells, P. A., Lynch, D F. J., Herrick, H. T. and May, O. E. 1937. Trans-
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Wells, P. A., Moyer, A. J., Stubbs, J. J., Herrick, H. T. and May, O. E. 1937.
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Gastrock, E. A., Porges, N., Wells, P. A. and Moyer, A. J. 1938. Gluconic
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Lvoff, S. and Loupizina, G. M. 1938. Effect of sodium fluoride on the formation
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Itaconic Acid

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Kojic Acid

Saito, K. 1907. tlber die Saurebildung bei A spergillus oryzae. Bot. Mag. (Tokyo)

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298 A MANUAL OF THE ASPERGILLI

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Oxalic Acid

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Miscellaneous Acids

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300 A MANUAL OF THE ASPERGILLI

Antibiotics and Toxins

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Waksman, S. A. and Geiger, W. B. 1943. The nature of the antibiotic substances
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Cook, A. H. and Lacey, M. S. 1944. An antibiotic from Aspergillus parasiticus.
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Hooper, I. R., Anderson, H. W., Skell, P. and Carter, H. E. 1944. The identity
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McKee, C. M., Rake, G. and Houck, C. L. 1944. Studies on Aspergillus flavus.

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Menzel, Arthur E. O., Wintersteiner, O. and Hoogerheide, J. C. 1944. The
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Chemistry of Mold Tissue

Norman, A. G., Peterson, W. H. and Houtz,R.C. 1932. I. Soluble Carbohydrate

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Biochem. Jour. 26: 1946-53. A. fischeri.
Pruess, L. M., Eichinger, E. C. and Peterson, W. H. 1934. III. Composition

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Jour. Amer. Chem. Soc. 56, No. 4, pp. 952-955.
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Prill, E. A., Wenck, P. R. and Peterson, W. H. 1935. VI. Factors Influencing

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Kroeker, E. H., Strong, F. M. and Peterson, W. H. 1935. VII. The Lipids of

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302 A MANUAL OF THE ASPERGILLI

Wenck, P. R., Peterson, W. H. and Greene, H. C. 1935. VIII. Innate Factors

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Wenck, P. R., Peterson, W. H. and Fred, E. B. 1935. IX. Cultural factors influ-
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Woolley, D. W. and Peterson, W. H. 1936. XI. Isolation of leucine and isoleucine

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Woolley, D. W. and Peterson, W. H. 1937. XII. Isolation of arginine, histidine,

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Woolley, D. W. and Peterson, W. H. 1937. XIII. Isolation of some monoamino-

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Bohonos, N., Woolley, D. W., and Peterson, W. H. 1942. XV. (Not numbered.)

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310 A MANUAL OF THE ASPERGILLI

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Physiology of the Aspergilli

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Steinberg, R. A. 1934. The so-called "Chemical Stimulation" of Aspergillus
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Steinberg, R. A. 1935. Nutrient -solution purification for removal of heavy metals
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Steinberg, R. A. 1935. The nutritional requirements of the fungus Aspergillus
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Gollmick, F. 1936. The influence of zinc, iron, copper and their combinations on
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Pigments and Coloting Substances

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Soil Tests for Mineral Deficiencies

Schlots, F. E., Smith, F. B. and Brown, P. E. 1932. Aspergillus niger as an indi-
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314 A MANUAL OF THE ASPERGILLI

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Variation

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Yuill, Edward. 1939. Two new Aspergillus mutants. Jour, of Botany, 174-175,
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Zahl, Paul A., Koller, L. R. and Haskins, C. P. 1939. The effects of ultra-
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Gossop, George Harold, Yuill, Edward and Yuill, John Lewis. 1940. Hetero-
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Simo, M. 1940. Experimentelle Untersuchungen uber die Wirkung von Radium
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Bedingung der Citronensauregarung durch Aspergillus niger Radium rasse.
Jour. Agr. Chem. Soc. Japan 16: 129.

Steinberg, R. A. and Thom, Charles. 1940. Chemical induction of genetic
changes in Aspergilli. Jour, of Heredity 31: 61-63.

Steinberg, Robert A. and Thom, Charles. 1940. Mutations and reversions in
reproductivity of Aspergilli with nitrite, colchicine and d-lysine. Proc.
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Whelden, Roy M. 1940. "Mutations" in Aspergillus niger bombarded by low
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Steinberg, R. A. 1942. Reversions in morphology of nitrite-induced "Mutants"
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Franklin Inst. 233:284.

31G A MANUAL OF THE ASPERGILLI

Vitamins and Growth Substances

Boysen-Jensen, P. 1931. Formation of a growth regulator by Aspergillus niger.
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Sakamura, T. and Yanagihara, T. 1932. The production of the growth substance
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Schopmeyer, H. 1932. Production of yeast-growth stimulants by molds on various
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Bunning, E. 1934. Growth and nitrogen assimilation in Aspergillus niger under
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Bernhauer, K. and Gorlich, B. 1936. The formation of vitamin C-like sub-
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Komarov, S. N. 1936. Methods of obtaining an antirachitic preparation from
the waste products of citric acid manufacture from the mycelium of Asper-
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Scheunert, A. and Schieblich, M. 1936. Vitamin production by Aspergillus
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Fukumoto, J. and Shinomura, H. 1937. Formation of Vitamin C-like substance
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Kitavin, G. S. 1939. Action of mercury salts on the formation of vitamin B2 in
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A. niger synthesizes biotin from biotin free media.

Miscellaneous Products
Alcohol

Molliard, M. 1916. Catalytic role of potassium nitrate in alcoholic fermentation

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318 A MANUAL OF THE ASPERGILLI

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Terrein

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Chapter XXIII
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324 A MANUAL OF THE ASPERGILLI

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Chapter XXIV
CHECK LIST OF SPECIES AND GENERA

GENERIC NAMES FOUND APPLIED
TO ASPERGILLI

Alliospora G. Pirn, in Proc. R. I.
Acad. 1883, and Jour. Bot. 1883,
p. 234; also Sacc. Syll. 18, No.
5216. 1906. A. Sapucaya rep-
resents a monotypic genus found
upon putrifying Lecythis Sapu-
cajo. From the description it
was some member of the A. niger
group. Saccardo's diagnosis
only seen 9

Ascophora. Ascophora nigrans was
vaguely assigned mostly to
Mucors but appears to have been
used by Raulin's group studying
the tannic acid fermentation
until Van Tieghem described
A. niger 9

Aspergillopsis Sopp, in Videnskaps-
selskapets Skr. I Mat.-Na-
turv. Kl. No. 11 , pp. 201-204, Taf .
XX, fig. 149. 1912. Sopp
probably had some members of
the A . ust us group 9

Aspergillopsis Spegazzini, in An.
Mus. Nat. Buenos Aires Ser.
3, 13: 434. 1911. The black
spored Aspergilli (essentially the
A. niger group) were regarded
as dematiaceous and assigned
to a new genus in that group.
Spegazzini 's genus has not been
accepted 8

Aspergillus Micheli, in Nova Plan-
tarumGenerap. 212, PI. 91. 1729.
Compare Link, in Obs. p. 16,
1809, and Corda in Icones
Fungorum 4: 31, Tab. VII, fig.

94. 1840 6

Subgenus — Microaspergillus Weh-
mer, in Monogr. 1901. (Mem-
oires de la Societe de Physique

et d'Histoire Naturelle de
Geneve, Tome XXXIII, Seconde
Parte, pp. 1-157. 1899-

1901) 16

Subgenus — Macroaspergillus Weh-
mer in Monogr. 1901. The
subgenera are not accepted
here 16

Cladosarum Yuill and Yuill, in
Trans. Brit. Myc. Soc. 22: 194-
200, Pis. 11-13. 1938. Re-
garded as a laboratory mutant
not encountered in nature. In
A. niger group. See 73

Dimargaris van Tieghem, in Ann.
Sci. Nat. 6 Ser. 1: 54. 1875.
The name Dimargaris sp. has
been seen upon living cultures in
one of the great laboratory col-
lections. These cultures were
obtained from dog and squirrel
dung. The organism so labeled
belonged to the Aspergillus
candidus group. The genus
Dimargaris (type D. crystalli-
gena v. Tieghem) is regarded by
Fitzpatrick as closely related to
Dispira and as a synonym of
Dispira by Zycha (Krypto-
gamenflora der Mark Branden-
burg. Pilze II. Mucorineae.
Band Via: 1-264, 114 figs.
1935). In any case, it is only
reported as a parasite of the
mycelia of the Mucorineae,
hence certainly was not an As-
pergillus 9

Diplostephanus Langeron, in Compt.
Rend. Soc. Biol. Paris 87: 343-
345. 1922. This genus is pro-
posed by Langeron to include
ascosporic Aspergilli (Sterig-
matocystis) with two series of

331

332

A MANUAL OF THE ASPERGILLI

sterigmata. A. nidulans Ei dam
is named as type. The proposal
is rejected here 8

Emericella Berkeley and Broome, in
Introd. Crypt. Bot. pp. 340-341,
fig. 76. 1857. See also: Pa-
touillard in Bui. Soc Myc.
France 7: 43-49, pi. 4, figs. 6-12.
1891. Based upon E. variecolor.
Syn. A . variecolor q.v 163

Euaspergillus Ludwig, in Lehrbuch
der niederen Kryptogamen p.
258. Stuttgart 1892. The
genus was proposed to cover the
sclerotium producing Aspergilli
such as A. niger, A. flavus, A.
ochraceus. The name has not
been accepted 8

Eurotium Link, in Obs. p. 31, Taf. 2,
fig. 44. 1809. Link described
Aspergillus glaucus as a conidial
mold and Eurotium herbariorum
for the ascosporic form, suppos-
ing them to be different fungi.
His genus Eurotium has been
widely accepted, but it is re-
jected here for reasons discussed
elsewhere. Forms listed as
Eurotium but having by descrip-
tion black perithecia are
excluded. No species with
aspergilloid conidial structures
has black (Dematiaceous) peri-
thecia 7

Inzengaea Borzi, in Jahrb. Wiss. Bot.
(Pringsheim) 16: 450-463, PL
19,20. (1884) 1885. Type sp.:

I. erythrospora Borzi: Syn.

A. variecolor q.v 165

Mucor used by Wiggers (Wichers)
in Primitiae Florae Holsaticae.
1780. (Republished in facsimile
Edition No. 23 by W. Junk in
1925). "Mucor herbariorum
sessilis luteus" is given as No.

1158 7

Rhodocephalus Corda, in Icones I:
21. 1839, and the figures given for
R. aureus in Icones III, Taf.

II, fig. 33 suggest A. terreus.
Sartorya Vuillemin, in Compt. Rend.

Acad. Sci. (Paris) 184: 136-317.
1927. Inadequately described
as an ascosporic phase of A.
fumigatus. It was not distribu-
ted in culture 9

Sterigmalocystis Cramer, in Viertel-
jahresschrift der Naturforchun-
gen Gesellschaft, Zurich 4: 325.
1859. Type: A. antacustica
Cramer. Cramer proposed to
include in his new genus all
aspergilloid species showing
both primary and secondary
sterigmata. S. nigra (A. niger)
was the organism under discus-
sion, hence the type species.
Many mycologists have agreed
with Cramer; others, such as
Fischer, Wehmer, and Thorn,
have rejected Sterigmato-
cystis 8

CHECK LIST OF NAMED SPECIES

S. acini-uvae Caballero, in Boletin
R. Soc. Esp. Hist. Nat. 28: 429.

1928 229

Syn. A. carbonarius fide Blochwitz,
or A. pulchellus fide Mos-
seray 229

S. aerea Bainier, in Bui. Soc. Bot.
France 28: 78. 1881. A.wentii
group 249

A. africanus Dur. et. M., in Fl. Alg.,
LP. 342. 1849. Not an Asper-
gillus.

A. ageni This name is cited by
Lindt, in Arch. Exp. Path.
Pharm. 25: 265. 1889, as taken
from Saccardo's Sylloge.
Search for this reference leads to
the conclusion that in this cita-
tion A. H ageni was made to read
A. ageni.

S. alba Bainier, in Bui. Soc. Bot.
France 27: 30. 1880. Some
member of A. candidus group. . 211

>S. alba-cyanogena Biourge, listed
among cultures in the Biourge
Collection. 1939.

S. alba-lutea Biourge, n. m., in MS.
p. 9, among the niveus series.

CHECK LIST OF SPECIES AND GENERA

333

A. alba-roseus attached to a culture
in the Bainier Collection and by
inference in the Biourge Collec-
tion as near A. niveus Blochwitz.
Thorn's No. 4640.490.

S. alba-sclerotifera Biourge, nomen
nudum, listed in his MS. as
related to A. okazakii. It
produces a violet or blue color
in reverse.

S. alba-sulphurea Biourge, nomen
nudum, listed in MS. as near
A. okazakii. It produces a blue
or violet reverse.

A. albidus Eichelbaum, in Verhand-
lungen d Naturw. Ver. Hamburg
3 Folge XIV; 35. 1906.
Syn. E. albidum Sacc, Syll. 22:
1254. 1913.

A. albidus Speg. A culture under
this label distributed by Biourge
represents a strain of A. niveo-
glaucus q.v 135

S. albo-lutea Bainier, in Bull. Soc.
Bot. France 27: 30. 1880. This
was a small pale yellow form, but
no further data were given and
it has not since been identified.
Apparently close to A. carneus.

S. albo-lutea Sartory, Sartory, and
Meyer, cited by Blochwitz, in
Ann. My col. 31: 73. 1933.
Some member of the A . candidus
group 211

A. albo-marginatus Biourge, in MS.
p. 2, as change of name from
Penicillium albo -marginatum,
figured only in the Penicillium
Monograph in La Cellule tome
XXXIII. 1 fasc. fig. 129. It
was some member of the A.
restrictus series, received and
studied as Thorn No. 4733.134a.
Near A . gracilis in the A . glaucus
group 139

S. albo-rosea Sartory, Sartory, and
Meyer, in Ann. Mycol. 28: 358-
359, PI. Ill, figs. 1-6. 1930.
Compare S. albo-rosea in
Blochwitz Ann. Mycol. 31: 75.
1933. A . carneus series 202

A. albus Wilhelm, Inaug. Diss.
Strassburg, p. 69. 1877. Prob-
ably A. albus Haller, S. alba
Sacc . , Monilia alba Persoon . In
A . candidus group 211

A. alliaceus Thorn and Church, The
Aspergilli, p. 163. 1926. See
A . wentii group 244

A. alternatus Berkeley, in Ann. Nat.
Hist., Ser. 1, Vol. 1: 262. 1838.
Not an Aspergillus.

A. alutaceus B. and C, in Grevillea
3: No. 25, p. 108. 1875. In the
A. ochraceus group, unidenti-
fiable to strain or series 281

S. ambari (alternative spelling,
ambaris) Beauregard, in Ann. de
Micrographie 10: 255-278, 1
plate. 1898. Cited without
description in Compt. Rend.
Hebdom. des Seances de la Soc.
de Biol. 28: Mai (1898). A.
vesicolor group 192

A. amoenus Roburg, in Hedwigia
70: 138-9. 1930. Distributed
by the Centraal -bureau voor
Schimmelcultures, Baarn. Re-
ceived by us on November 20,
1933, and determined as a
member of the A. gracilis series,
but the description allies it with
A. versicolor.

A. amstelodami (Mangin) Thorn and
Church, in The Aspergilli, p.
113. 1926; also Thorn and
Raper, U. S. D. A. Misc. Publ.

426, pp. 22-26. 1941 122

Syn. E. amstelodami Mangin, in
Sci. Nat. Bot. Ser. 9, 10: 360-
361. 1909. A. glaucus group.... 122
Incorrectly spelled A . amsterodami
in Nakazawa, Jour. Agr. Chem.
Soc. Japan 10(2): 1934.
Syn. E. repens var. amstelodami
Vuill., in Soc. Mycol. de France,
Bui. Trimest. 36: 131. 1920.
var. alophote Duche, cited by
Dodge in his Med. Mycol. p. 630.
1935. Ascospore lacks the crest
of the species.

334

A MANUAL OF THE ASPERGILLI

A. anomalus Mosseray, in La Cellule
XLIII: 248-249, pi. 4, figs. 107-
112. 1934. Member of A . niger
group 234

S. antacustica Cramer, in Vrtljschr.
Naturf. Gesell. Zurich, Jahrg.
4, Heft. 4: 325. 1859. In A.

niger group 239

Syn. A. phoenicis in A. niger
group 222

A. archaeoflavus Blochwitz, in Ann.
My col. 31(1/2): 73-83. 1933.
The type culture (Thorn No. 250-
5346) was too close to A. wentii
for satisfactory separation 247

A. archiflavipes Blochwitz, in Ann.
Mycol. 32(1/2): 84. 1934. A
form of A . flavipes characterized
by deep brown to red shades of
color in the mycelium 181

A. argentinus Spegazzini, in Rev.
Agr. Veter. LaPlata p. 245.
1896. A conidial form in the
A. glaucus group; not identifi-
able.

A. argillaceus was distributed but
not described by Biourge. It
appears to be a non-ascosporic
strain of the A. repens series — 111

A. atro-fuscus Mosseray, in La
Cellule XLIII: 269-270, pi. 4,
figs. 126-129. 1934. Member of
A . niger group 235

A. atropurpureus Zimmermann, in
Centbl. Bakt. [etc.] Abt. 2, Bd.
8, No. 5, p. 218. 1902. A. niger
group 226

A. atropurpureus Blochwitz, in Ann.
Mycol. 32(1/2): 86. 1934.
Blochwitz proposed this name to
cover purple-brown series in A.
niger group 225

A. atro-ruber Estienne

Syn. Penicillium roseo-cinnabari-

num Biourge, in La Cellule 33:

fasc. 11, 319-321, PI. XVII, fig.

97. 1923'.

Syn. Physomyces heterosporus

Harz, fide Biourge.
Syn. S. atro-rubra (Estienne)
Biourge MS.

A. atro-violaceus Mosseray, in La
Cellule XLIII: 268-269, pi. 4,

figs. 122-125. 1934 235

Blochwitz in Ann. Mycol. 33: 240.
1935, identifies this with A.

violaceo-fuscus Gasperini 231

A. atrovirens Karst, in Symb. 26: 28;
Sacc. Syll. 10: 524. Inadequate-
ly described for identification.
A. aurantiacus Berkeley, in British
Fungi fasc. IV: 1843. Cited
by Montagne in Ann. Sci. Nat.
Bot. 3. Ser., 12: 299. 1849.
Syn. N ematogonium aurantiacum.
S. aurea Greco, in Origine des
Tumeurs et Mycoses Argentines,
Buenos Aires, pp. 671-694, figs.
418-428. 1916. Not since recog-
nized. Some A . ochraceus strain.
A. aureoglaucus Roburg, in Hedwigia
70: 137. 1930. Culture some
strain of A. repens contributed
by Roburg. Noted as some
member of A. glaucus group
by Blochwitz in Ann. Mycol.
33: 240. 1935.
A. aureus Berkeley, in English Flora
5: 346. 1836. The golden
yellow, elliptical conidia re-
ported by Berkeley suggest
A. citrisporus but actual identi-
fication from the description is

impossible 219

A. aureus Nakazawa, in Inst. Gov't.
Res. Formosa Rept. Vol. 1.

1907 219

A. aureus varieties described by
Nakazawa, Simo and Watanabe
in Jour. Agr. Chem. Soc. Japan
No. 144, 1936 (In Japanese).

var . acidus 220

var . brevis 220

var. minor 220

var. murinus 220

var. pallidus 220

comparison of figures and measure-
ments given convince the authors
that these varieties could not be
safely identified by descriptive
data.

CHECK LIST OF SPECIES AND GENERA

335

S. auricoma Gueguen, in Bull. Soc.
Mycol. France 15: 171-187, figs.
1-48. 1899. In A. ochraceus

group 276

A. auriculaire Moquin-Tandon, in
Elements Bot. Med. 2 ed., p. 466.
1866. This name was not
accompanied by adequate de-
scription to separate the form
intended from other species
which are occasionally found in
the ear.
.4. ariarius Peck, in N. Y. State
Mus. Rept. (1890), 44: 120.
1892. A strain of A. fumigatus

isolated from a canary 151

A. avenaceus George Smith, in Brit.
Mycol. Soc. Trans. 25: 24-27,

PI. 1, figs. 1-3. 1943 246

A. awamori Nakazawa, in Inst, of
Gov't. Research, Formosa Rept.
1: 1907, and 2: 1912. A member

of the A . niger group 220

A. awamori varieties described by .
Nakazawa, Simo, and Watanabe
in Jour. Agr. Chem. Soc. Japan,
No. 144. 1936. (In Japanese).

var. ferrugineus 220

var. furneus 220

var.fuscus 220

var. minimus 220

var. piceus 220

No doubt the describers had a

series of industrially significant

strains but it is doubtful if others

could identify them from the

description and figures given.

A. awamori Usami, in Myk. Cen-

tralbl. 4: 193. 1914. Species

recognized by Mosseray in La

Cellule 43: 264. 1934. With A.

awamori Nakazawa cited as a

synonym.

A. bainieri Mosseray, in Ann. Soc.

Sc. Bruxelles 54, ser. B, p. 79,

1934 233

Syn. A. longobasidia Bainier fide
Mosseray in La Cellule XLIII

(2): 227. 1934 233

A. barbae Castellani. Cited by
Sartory, A., in Champignons

parasites de l'homme et des
animaux, p. 595. 1922. This
organism was found in the beard
of a native of Uganda and again
from Ceylon. The only further
information given is conidia 4 to
5ju, globose, dark brown.

S. basidiosepla Sartory, Sartory, and
Meyer, in Ann. Mycol. 27: 317-
320. PI. VII. 1929. A variant
of the A . candidus group 211

A. batatae Saito, in Centralb. f. Bakt.
2, Abt. 18(1/2): 34. 1907. In
the A . niger group 225

A. belfanti Carbone, in Atti d. Inst.
Bot. Univ. Pavia Serie II. Vol.
XIV: 63, fig. 11. 1914. Prob-
ably in A. glaucus group.

A. (Sporodinia) bellomontii Mon-
tagne, in Ann. Sci. Nat. Bot. Ser.
4, V. 12: 181-182. 1859. Noted
as related to A . maximus (Sporo-
dinia). Not an Aspergillus.

S. bicolor J. Ray, in Rev. Sc, Ser. 4,
V. 8: 176-177, 193-212. Lille,
1897. Probably in A. sydowi
series 192"

A. biourgei Mosseray, in La Cellule
XLIII: 241-242, pi. 4, figs. 81-
85. 1934. One of the A. niger
group 234

S. blanc-jaune Bainier, nomen
nudum. A culture so labeled
from the Bainier Collection rep-
resents a diminutive member
of the A. candidus group 211

A. blochwitzii Biourge, nomen
nudum, listed in Biourge MS
(p. 3) among the A. clavatus
group as a synonym of A. cla-
vatus var. gigantea Blochwitz.

A. boedijni Blochwitz, in Ann.
Mycol. 32: 83-89. 1934.
Syn. A. terreus var. boedijni 197

E. bonariense Speg., in Anal, de la
Soc. Cient. Argen. 10: 177.
1880. Some member of the A.
glaucus group but data are
inadequate.

Coremium Borzianum Saccardo.

See A . variecolor 165

336

A MANUAL OF THE ASPERGILLI

A. bouffardi Brumpt (1905). Cited

by Castellani and Chalmers in

Man. Trop. Dis. p. 805. 1913.

Syn. Madurella Bouffardi (Brumpt)

Dodge. Med. Myc. p. 685. 1935.

A. Brodeni (Mattlet) Dodge, in
Dodge Med. Mycol., p. 635.

1935 170

Syn. S. Brodeni Mattlet, in Ann.
Soc. Beige Med. Trop. 4: 167-
171, figs. 1, 2. 1924. Probably
A. unguis.
var. V ancampenhouti (Mattlet)
Dodge, in Dodge Med. Myc,
636. 1935.

A. bronchialis Blumentritt, in Ber.
Deut. Bot. Ges. 19: 442-446, pi.
22, figs. 1-6. 1901 ; also ibid. 23:
419-127, pi. 19, figs. 1,3, 6, 7,8,
19, and 23. 1905. In the A.
fumigatus group 151

A. brunneo-fuscus See, in Les
Malades du papier pique Paris,
p. 29. 1919. Unidentified ex-
cept as a member of the A.
glaucus group.

A. brunneo-virens Delacroix, in Bui.
Soc. Myc. France 13: 120, with
text figure. 1897. Unidentified
except as a member of the A.
glaucus group.

A. brunneus Delacroix, in Soc.
Mycol. Bui. France 9: 185, PI.
XI, fig. III. 1893. Delacroix
described this as the conidial
stage of A. echinulatus q.v 131

S. Buntingiana Biourge, nomen
nudum, listed in Biourge 's MS. as
applied to a culture of a member
of the A. candidus group re-
ceived from Bunting.

A. Buntingii Mosseray, in La Cellule
XLIII: 236-238, pi. 4, fig. 91-95.
1934 . Member of A . niger group

in Biourge Collection 1939 233

A. butyracea (Bainier) n. comb 282

Syn. S. butyracea Bainier, in Bull.
Soc. Bot. France 27: 29. 1880.
C. Roumeguere's Fungi Gallici
Exsiccati No. 995 is recorded

as Bainier's material. In the
A . ochraceus group 282

A. byssoides Sprengel, in Sys. Veg.,
ed. 16, V. 4: 541. 1827. A
fungus on rotting paper with
globose fuscous heads but not
identifiable.

A. cacao. This name appeared upon
a culture in the Bainier collec-
tion (Thorn No. 4640.397) and
has been contributed also by
Pribram (4777.4). The organ-
ism is a strain of A. tam-
arii 256

A. caesiellus Saito, in Jour. Coll.
Sci. Imp. Univ. Tokyo 18: 49,
PL III, fig. 14. 1904. Re-
garded by Neill (in Royal Soc.
New Zealand Trans. 69: 242.
1939) as A . restrictus G. Smith. . . 141

A. caespitosus n. sp. Raper & Thom,
in Mycologia 36: 563-565, fig. 3.
1944. A member of A . nidulans
group 166

A. calyptratus Oudemans, in Arch.
Neerl, II. 7: 283, pi. 13. 1902.

See: A. fumigat us group 151

var. italicus Ferraris, in Ann. My-
col. 10: 294. 1912.

S. cameleo Sartory, Sartory, and
Meyer, in Ann. Mycol. 29: 360-
361, PI. III. 1930. Some vari-
ant of A . sydowi 186

S. Candida Sacc, in Michelia 1, p. 91.
1877. In the A. candidus
group.

S. candidula Bainier, in Sacc. Syll.
4: 73. 1886.
Syn. S. Candida Bainier, in Bui.
Soc. Bot. France 27: 30. 1880.
A member of the A. candidus
group.

A. candidus Link, in Obs. p. 16.

1809 207

var. thermophilus Nakazawa et al.,
in Jour. Agr. Chem. Soc. Japan
88: 17. 1932, (in Japanese)
cited Blochwitz Ann. Mycol.
33: 244. 1935.

A. capitulo pullo Haller, in Historia
Stirpum Indigenarum Helvetiae

CHECK LIST OF SPECIES AND GENERA

337

Inchoata, etc. 1768. Appar-
ently taken from Micheli 1729.

S. carbonaria Bainier, in Bui. Soc.
Bot, France 27: 27-28. 1880.
See A. carbonarius (Bainier)
Thorn. In .4 . niger group 229

A. carbonarius (Bainier) Thorn, in

Jour. Agr. Res. 7: 12. 1916 229

A. carbonarius sen aler Meis and
Parascandalo, in Gaz. Ospedali
16: 769-772. 1895. Cited by
Dodge, in Med. Myc. 679, as not
an Aspergillus.

A. carncolus Sacc, in Michelia 1, p.
77. 1877. And Fungi italici
No. 18. In A. glaucus series.

A. carneus (van Tieghem) Bloch-
witz, in Ann. Mycol. 31(1/2):

81. 1933 201

Syn. Sterigmatocystis carnea van
Tieghem, in Bull. Soc. Bot.
France 24: 103. 1877. See also
Saccardo Sylloge 4: 74, and
Wehmer's Monograph (Mem.
Soc. Phys. His. Nat. Gen. pp.
1-157). 1899-1901. Species in

A . terreus group 201

var. subglobosa Blochwitz, in Ann.

Mycol. 33: 243. 1935.
var. opaca Blochwitz, in Ann.
Mycol. 33: 249. 1935.

A. carnoyi Biourge, descr. Thom and
Raper in U. S. Dept. Agr. Misc.
Pub. 426, p. 34-5. 1941 134

S. castagnei Biourge, undescribed
culture in the Biourge Collec-
tion, listed as near A. carbo-
narius in the A. niger group.

A. castonea Patterson, in Bui. Torrey
Bot. Club 27: 284. 1900. In A.
tamarii series.

A. cellulosae Hopffe, in Centralb. f.
Bakt., etc., 1 abt., 83: 531-37.
1919.
Syn. A.fumigatus 151

A. cervinus Massee, in Kew Misc.
Bui. 4: 158. 1914. Neill con-
siders A. gratioti Sartory to be a
synonym but that is not
probable.

A. chevalieri (Man^in) Thom and
Church, in The Aspergilli, p.
Ill, 1926.
Syn. E. chevalieri Mangin, in Ann.
Sci. Nat, Bot. Ser. 9, 10: 361-
362, fig. 12. 1909. In the A.
glaucus group 118

A. chevalieri var. intermedins Thom
and Raper in U. S. Dept. Agr.
Misc. Pub. 426, p. 21, fig. 8B.
1941 121

A. chevalieri var. rnultiascosporus
Nakazawa, Takeda, Okada, and
Simo, in Agr. Chem. Soc. Japan,
Jour. 10: 135-192. 1934. Re-
ceived from Baarn and appears
as NRRL No. 88. A strain of
A . chevalieri 120

E. chilense Montagne, in Syll. Crypt.
No. 919, Fl. Chil. VII, p. 476;
cited by Saccardo Syll. 1, p. 27.
In the A. glaucus group.

S. chlorina Cooke and Massee, in
Grevillea 18: 7. 1889. Proba-
bly a nonascosporic strain of the
A. glaucus group.

A. chrysospermum Thaxter, nomen
nudum, attached to a culture
later shown to be A. citrisporus
Van Hohnel, q.v.

A. chungii Shih, in Lingnan Sci.
Jour. 15(3): 378. 1933. In the
A . flavus group 267

A. churchii Mosseray, in La Cellule
XLIII: 242-244, pi. 4, figs. 76-80.
1934. Representative of the A.
niger group. In the Biourge
Collection 1939 234

A. cimmerius Berkeley and Curtis,
in Grevillea 3: 108. 1875. This
specimen was identified by Dr.
Farlow (Bibliographical Index
I. pt. 1, page 277) as Periconia
chlorocephala, Fres.

A. cinerescens Bainier and Sartory,
in Bui. Soc. Mycol. France 27:
98-104, pi. Ill, figs. 6-12. 1911.
In the A. glaucus group.

A. cinereus Spegazzini, in Anal. Soc.
Cient. Argen. 10: 162-163. 1880.

338

A MANUAL OF THE ASPERGILLI

Not identifiable; believed in the
A. glaucus group.

A. cinnamomeus Schiemann, in
Ztschr. Induk. Abstain, u.
Vererbungslehre, Bd. 8, Heft
1/2: 1-35, 16 figs. 2 pi. (1 col.).

1912 223

Syn. A . niger mut. cinnamomeus . . . 223

A. cinnamominus (Weiss) Dodge, in
Dodge Med. Myc., p. 627.

1935 200

Syn. S. cinnamominus Weiss,
in Ann. Parasitol. Hum. Comp.
8: 189-193, 5 figs. 1930. See
A . terreus 200

A. circinatus Mangin, in Bui. Soc.
Mycol. France 15: 223, PI. XI,
figs. 5, 6, 7. 1899. Not recog-
nizable— probably not an Asper-
gillus.

A. citricus (Wehmer) Mosseray, in
La Cellule XLIII: 262-263, pi.
4, fig. 105-106. 1934. Memberof
A. niger group. In Biourge's
Collection 235

A. citrino-niger Mosseray, in La
Cellule XLIII: 231-232, pi. 1,
fig. 7. 1934. Member of A.
niger group. In Biourge's Col-
lection only 233

A. citrisporus von Hohnel, in Sit-
zungsber. Kais. Akad., Wiss.
Wien, Math- Naturw. Kl.
Ill, Abt. I: 987. 1902. In the
A. tamarii group 251

A. citromyces E. Ruppel, in J. Pharm.
Belg. 19: 63-8. 1937. C.A. 31:
6687.

A. clavatus Desm., in Ann. Sci.
Nat. Bot. Ser. II, 2: 71, pi. 2,
fig. 4. 1834 92

A. clavatus var. gigantea Blochwitz.
Cited by Biourge in unpublished
MS.

A. clavatus Desm. mut. gigantea
Blochwitz, in Ann. Mycol.
XXVIII. (3/4) : 1920 is used by
Blochwitz on several occasions.. . 97

A. clarellus Peck, in N. Y. St. Mus.
Rept. 34: 44, pi. 2, figs. 1-5.
1881.
Syn. A . clavatus Desm 98

S. coerulea Blochwitz, listed in
Biourge's MS. as one of the blue
Aspergilli (A. sydowi).

A. concentricus Castellani, in Trans.
Int. Derm. Cong. 6: 667-671, pi.
49, 50. 1907.
Syn. Epidermophyton concentricum
(Blanchard) Castellani and
Chalmers.

A. condylomatae Greco. A name
attached to a pathogen without
data for identification.

A. conicus Blochwitz published by
E. Dale in Ann. Mycol. 12: 38.
1914. Described as a Penicil-
lium in Ann. Mycol. 10: 465.
1912. In the A. restictus
group 140

A. conoideus Sprengel, in Sys. Veg.
ed. 16, V. 4: 541. 1827. This
is cited as Byssus conoidea Mull.
in Flora Danica, Taf . 897, fig. 2
which is a myxomycete, hence
this name may be dropped.

A. cookei Sacc, in Sylloge Fung. IV:
71. 1886.
Syn. A. mucoroideus Cooke, in
Grev. 12: 9. 1883. Some mem-
ber of the A. glaucus group.

E. coriorum Wallr., in Compendium
Flora Germania 4: 330. 1833.
Syn. E. repens DeBary fide Man-
gin. Not recognizable.

S. corolligena Massee, in Bui. Royal
Bot. Gard. Kew. No. 1, p. 5.
1910. Not recognizable.

S. coronata v. Tieghem, in Bull. Soc.
Bot. France 24: 103. 1877.
Unidentifiable.

S. coronella Costantin, in Muced.
Simples p. 34, fig. 2, 1888. Uni-
dentifiable.

A. crocatus Berkeley and Curtis.
Specimen only in the Curtis
Collection. This type specimen
has been identified as Chon-
dromyces and is so reported in
Farlow's Bibliographical Index
I. pt. 1, page 277.
A. cucurbitaceus in the Curtis Collec-
tion (Harvard). Was identified
by Farlow as Choanephora.

CHECK LIST OF SPECIES AND GENERA

339

A. curtisii Berkeley, in Ilavenel
Fungi Carolinense fasc. IV.
1855. Farlow identified this
in his Bibliographical Index as
Rhinotrichum curtisii.

S. cyanea Bainier, nomen nudum, in
Biourge's list of the nidulans
group.

A. cyaneus (Mattlet) Dodge, in
Dodge Med. My col. p. 636, 1935.
Syn. S. cyaneus Mattlet, in Ann.
Soc. Belg. Med. Trop. 6(1): 32.
1926. In A . sydowi series 186

A. cyanogenes Biourge, nomen
nudum. One of the A. conicus
strains in A . restrictus series 140

S. dasytricha Ell. and Ev., in Jour.
My col. 2: 104. 1886. Sacc.
Syllog. X: 525. Not an Asper-
gillus.

A. delacroixii (Sacc.) Thorn and
Church, in The Aspergilli, p. 190.

1926 282

Syn. S. delacroixii Sacc, in Sylloge

10:527 282

Syn. S. ochracea Delacroix, in Bui.
Soc. Mycol. France 7: 109, pi.
VII, fig. f. 1891 282

A. delacroixii Sacc. and Sydow, in
Sylloge Fungorum 14: 1044.
Syn. A. olivaceus Delacr., in Bui.
Soc. Mycol. France 13: 118.
1897. Some unidentifiable

member of the A. glaucus group.

A. densus Mosseray, in La Cellule
XLIII: 232-234, pi. 4, figs. 99-
102. 1934. In the Biourge
Collection 233

A. depauperatus Petch, in Trans.
Brit. Mycol. Soc. XVI. p. 244,
text fig. 1931. Parasite upon
Lepidosaphis ulmi on Hawthorn
in England, also on Aspidiotus
in Ceylon. As described: coni-
diophores up to 50m by 2.5 to 3.5m,
broadening to a fertile apex 4 to
5/x in diameter with a few chains
of conidia attached directly to
the vesicular surface; conidia 2
to 4m by 1.5 to 2.5u. Possibly
some strain of the A. restrictus
series not far from A. gracilis.

A. derxii Biourge, nomen nudum,
listed among the A . versicolor
group in unpublished MS. and
represented in his collection in
1939.

E. desmazieri Castagne, PI. Mars.
II, Sup., p. 56. 1851. See also
Berlese, Fungi Moricolae Ap-
pendice p. 11. 1889. As de-
scribed by Berlese, this was a
perithecial form probably be-
longing to the A. glaucus group,
but not sufficiently described to
identify.

A. desseyi Speg., in Physis., Rev.
Soc. Argentina Cien. Nat. VIII:
115-117, 1 fig. 1925. Named
A. dessyi Speg., in Rev. Appd.
Mycol. 4: 542. 1925. Some
form near A . fumigat us 151

A. dierckxii Biourge, nomen nudum,
on culture distributed by
Biourge; a strain of the A.
repens series. See Thorn and
Raper U. S. D. A. Misc. Publ.
426, p. 12. 1941 107

A. diplocystis (Sartory, Sartory,
Hufschmitt and Meyer) Dodge,
in Dodge Med. Mycol. p. 625.

1935 121

Syn. E. diplocyste Sartory,
Sartory, Hufschmitt and Meyer,
in Compt. Rend. Soc. Biol. 104:
881-883. 1930. The authors
describe a mold with conidial
head near A. nidulans and the
ascospores of A. chevalieri 121

E. diplocystis B. and Br., in Jour.
Linn. Soc. (London), Bot. 14:
55-56, Tab. 10. 1875. Neither
description nor figure would
identify this with any definite
Aspergillus, although the speci-
men may have been the perithe-
cia of some member of the A.
glaucus group.

S. dipus Ferdinandsen and Winge, in
Bot. Tids. 30: 220, fig. 6. 1910.
Not separable from other large-
spored forms in this section of
the A . niger group 228

340

A MANUAL OF THE ASPERGILLI

-4. disjunctus Bainier et Sartory, in
Bui . Soc . My c . France 27 : 346-368,
pi. X-XI. 1911. A member of
the A. glaucus group. SeeA.echi-
nulatux 133

A. diver sicolor Waksman, in Soil
Science 2: 126. 1916. The refer-
ence to an organism by Waksman
under this name was a typo-
graphical error for A. versicolor.

S. dubia (B. & Br.) Sacc, in Fungi
italici pi. 902, and Sylloge 4: 72.
1886.
Syn. A. dubius Corda, in Berkeley
and Broome, in Ann. Nat. Hist.
2 Ser., 7: 100 (No. 520). 1851.
Not thus far recognized.

A. dubiosus Lindau, in Rabh. Krypt.
Fl. 8: 151. 1907. In the .4. can-
didus group.

A. eburneus Biourge, nomen nudum,
was attached to a culture re-
ceived from Biourge and re-
corded as NRRL 515. It is
accepted as A. niveus 202

A. echinosporus Sorokin. Abst. by
Busch, in Ztschr. f. Pflanzen-
krankheiten 3: 155. 1893. Ref.
in Sacc. Sylloge 11: 592. 1895.
The description suggests a Hap-
lographium.

A. echinulatus (Delacr.) Thorn and
Church, in The Aspergilli , p. 107.

1926 131

Syn. E. echinulatum Delacr., Soc.
My col. de France, Bui. Trimest
9: 266, PI. XIV, fig. III. 1893.
Syn. E. verruculosum Vuill., in Soc.
Mycol. de France, Bui. Trimest.
34: 83. 1918. In A. glaucus
group 131

A. effusus Tiraboschi, in Ann. di Bot.
7 (fasc. 1): 16. 1908. See also
Thorn and Church, in Am. Jour.
Bot. 2:109-110. 1921. Describ-
ed originally from rotten corn
(Zea Mays); belongs to the A.
flavus-oryzae group 267

A. elatior Mosseray, in La Cellule
XLIII: 253-255, pi. 3, figs. 29-32.
1934. In the Biourge Collection. 234

A. elegans Gasperini, in Atti. Soc.
Toscana Sci. Nat. Pisa, Mem. 8:
(fasc. 2): 328. 1887. In A.
ochraceus group 281

E. epixylon Kunze and Schm. (D.
Schw. No. 83; Wallr. Fl. Crypt.
No. 2078). In Compendium
Florae Germanicae 4: 331 . 1883.
This was evidently some member
of the A. glaucus group.

A. erythrocephalus Berkeley and
Curtis, in Jour. Linn. Soc.
[London], Bot. 10: 362. 1869.
In A. tamarii group 257

Inzengaea erythrospora Borzi, in
Jahrb. Wiss. Bot. (Pringsheim)
16: 450-463, pis. 19-20. 1884.
Manifestly A. variecolor 163

A. exiguus Hann.

Syn. A. conicus fide Blochwitz, in
Ann. Mycol. 31: 73. 1933.

S. ferruginea Cooke, in Grevillea
VIII. (1879), 95., in Jour.

Quekett Mic. Club 2 Ser. 2: 139.
1885. Not an Aspergillus. Re-
published in Veg. Wasps and
Plant Worms, p. 184. 1892; see
also Petch in Trans. Brit. Myc.
Soc. XVI, p. 72. 1931, who ex-
amined type specimens as rust-
colored patches on pupae; co-
nidophores hyaline smooth, 12m
diam., vesicle globose 50m ; pri-
mary sterigmata 30 by 4 to 12/*,
secondary 14-18 by 7-9m, conidia
from 9 by 5m to 6 to 9m subglo-
bose, brown, coarsely rough; a
brown member of the A. niger
group.

A. ferrugineus Fuckel, in Fungi
rhenani No. 157, also Symbolae
Mycologicae in Jahr. d. Nas-
sauischen Vereins fur Natur-
kunde Jahrg. XXIII and XXIV:
358. 1869-70. Not identified.

A. ferrugineus Link, in Sp. Plant, Ed.
4, 6: pt. 1, p. 68. 1824. Also in
Fries. Sys. Myc. 3, p. 387. 1829.
Both authors seem to have had
members of the A . glaucus group.

A.ficuum (Reich.) Hennings 225

CHECK LIST OF SPECIES AND GENERA

341

Syn. Sterigmatocystis ficuum
(Reich.) P. Henn., in Hedwigia
34, Heft 2: 86. 1895.
Syn. Istilago ficuum Reichardt,
in Verhandl. K. K. Zool. Bot.
Gesell. Wien. 17: 335. 1867.
A culture distributed under this
name (Thom 142) is a rapid ox-
alic acid producing organism.
In the A. niger group.

A.fiemonthi Sopp, cited as a name on
a culture by Biourge in MS. p.
19, in the A. flavus group.

A. fimetarius Peck, in N. Y. State
Mus. Bot. Rept. 42, p. 128.
1889. Probably .4.. candidus
group.

A. fimeti Sacc, in Michelia 2: 543.
1882. Inadequately described.

A. fischeri Wehmer, in Centralb. f.
Bakt. II abt. 18(12/15): 390-392,
figs. 5. 1907. In the A. fumi-

gatus group 151

Syn. A. fumigatus-ascosporic, see
Thom and Church, in Am. Jour.
Bot. 5: 91-92. 1918.

A. flavaureus Biourge, nomen nu-
dum, listed in his MS. as a mem-
ber of the ^4. flavus-oryzae group
with yellow-orange reverse, as
shown by a culture in his collec-
tion.

A. flavescens Wreden, in Compt.
Rend. Acad. Sci. Paris 65: 368.
1867. Also St. Petersb. Med.
Zeitschr. 13: 133. 1867. Iden-
tifications have not been satis-
factory. Possibly in A. nidu-
lans group.

A. flavidus Berkeley and Broome, in
Jour. Linnean Soc. (London),
Bot. 14: 101. 1875. Cited by
Saccardo in Fungi of Ceylon,
London 1871-73, No. 913 S.
Probably a member of the A.
flavus group but not more closely
identifiable.

A. flavipes (Bainier and Sartory)
Thom and Church, in The Asper-
gilli, p. 155. 1926 179

Syn. S. flavipes Bainier and Sar-
tory, in Bui. Soc. Mycol. France
27: 90-96, pi. Ill, figs. 1-6.
1911 179

A. flavo-viridescens Hanzawa, in
Journ. Coll. Agr. Tohoku Imp.
Univ. Sapporo 4: 232-3, PI. 21,
figs. 1-4. 1911. In A. versicolor
group 192

A. flavus Link, in Obs. p. 16, 1809.
(H. F. Link Observations in
Ordines plantarum naturales,
Gesellschaft Naturforschender
Freunde zu Berlin, Magazin 3,
1809 — commonly cited Link
Obs.) 263

A. flavus forma Maydis Ciferri, R.,
in Ann. Mycol. 20: 46. 1922.

A. flavus var. japonica. cited by
Blochwitz in Ann. Mycol. 31: 74.
1933, without description.

A. flavus var. viridis Blochwitz, in
Ann. Mycol. 32: 86. 1934. Is
vaguely designated as a non-
tropical strain apparently dupli-
cating A. parasiticus Speare. . . 267

A. flavus mut. rufa Blochwitz, in
Ann. Mycol. 27(3/4) : 201 . 1929.
A mutation which shows grada-
tions to pure brown from the
green form.

A. flavus mut. fusca Blochwitz, in
Ber. d. Bot. Ges. 48: 576. 1928.

Eurotium Aspergillus flavus DeBary
and Wor. Terminology em-
ployed by DeBary and Woronin
(Beitr. z. Morph. u. Physiol, d.
Pilze, III Reihe, p. 380. 1870)
to designate relationship of the
asexual species, A. flavus, with
species characterized by a per-
fect stage and included in Euro-
tium 263

A.foetidus n. sp 219

Syn. A. aureus Nakazawa. A
black Aspergillus with a strong
musty odor 219

A. fonsecaeus n. sp.

Syn. S. fusca Bainier, in Bui. Soc.
Mycol. France 27: 29, pi. 1, fig. 5.
1880. A black Aspergillus on

3-42

A MANUAL OF THE ASPERGILLI

rubber, from Fonseca in Rio de
Janeiro 227

A. fontoynonti Gueguen, in Archives
de Parasitologic 14: 177-192, 2
plates, 36 figs. 1910. See also
Compt. Rend. Soc. Biol. Paris
66:1052. 1909. The figures and
description place this form un-
questionably in the A. glaucus
group 147

A. freisingianus Biourge, nomen
nudum, listed with the A. clava-
tus group as "Tres liquifiant,
comme celle de Blochwitz." In
the Biourge Collection 1939.

E.fructigenum Link, in Sp. PI. Ed. 4,
6: 80. 1824. Some unidentifi-
able member of the A. glaucus
group.

S. fuliginosa Bainier, in Bui. Soc.
Bot. France 28: 79. 1881. Some
variety of A. niger.

A. fuliginosus Peck, in Bui. Buffalo
Soc. Nat. Sci. 1: 69. 1873. Also
in Ann. Rep. N. Y. St. Mus. Nat.
Hist. 26: 79. 1874. Some
strain of the A. niger group.
Mosseray applied it to a culture
in Biourge's Collection 233

E. fulvescens Cooke, in Grevillea 8:
11. 1880.
Syn. Badhamia fulvescens Cooke.
Change of generic designation
only, from the same material.
Not identifiable from descrip-
tions.

A. fulvus Montagne, in Ann. Sci.
Nat. Bot. Ser. 3, Vol. XII: 298.
1849.
Syn.<S./uZraSacc.Syll.4:74. Not
identifiable. Described from
diseased silk worms in Southern
France, but not since identified.

A. fumaricus Wehmer, in Ber. Deut.
Chem. Gesell. 51(14): 1663-68,
figs. 1-6. 1918. A unique mem-
ber of the A . niger group 227

Sartorya fumigata Vuillemin, in
Compt. Rend. Acad. Sci. (Paris)
184: 136-137. 1927. Possible
synonym of A. fischeri q.v 153

A. fumigatoides Bainier and Sartory,
in Bull. Soc. Mycol. France 25:
112, pi. 5. 1909. Compt. Rend.
Soc. Biol. [Paris] 66: 22. 1909.
See also Sartory, A., Champig-
nons parasites de l'homme et des
animaux, p. 573. 1922 153

Syn. E. fumigatoides (B. & S.)
Sacc.SylI.22:1255. 1913. Prob-
ably close to A. fischeri 153

A. fumigatus Fresenius, in Beitrage
zur Mykologie, p. 81, pi. 10, figs.
1-11 Frankfurt. 1850-53 148

var. Alpha, Sion and Alexan-
drescu, in Compt. Rend. Soc.
Biol. 64: 288-289. 1908 151

var. cellulosae Baumli, cited by
Sartory, Sartory, and Meyer, in
Compt. Rend. 203: 1289-91.
1936. See C. A. 31: 2249. 1937.

Mut. helrola E. Yuill, in Jour. Bot.
(London) 1939. p. 174. A
buff-colored mutant which ap-
peared in culture 150

var. magnus Nakazawa, Simo, and
Watanabe, in Jour. Agr. Chem.
Soc. Japan No. 144. 1936. (In
Japanese.)

var. minimus Surtory, in Bui.
Acad. Med. Paris Ser. 3, 82: 304.
1919 151

var. tumescens Blumentritt, in Ber.
Deut. Bot. Gesell. 23: 419-427,
PI. 19, fig. 5, 6, 18-21. 1905.... 151

var. subglobosus Blochwitz, in Ann.
Mycol. 33: 244. 1935.
Aspergillopsis fumosus Sopp, in
Videnskapselskapets Sks. I
Mat.-Naturv. Kl. No. 11, pp.
202-204, Taf. XX, fig. 149. 1912.
Not identified but suggested as
some strain near A. ustus.
A. fungoides Greco, in Origine des
tumeurs et mycoses Argentines,
Buenos Aires pp. 505-509, figs.
297, 298, 299. 1916. The de-
scription is inadequate for iden-
tification.
S. fusca Bainier, in Bull. Soc. Bot.
France 27: 29, pi. I, fig. 5. 1880.
Compare Roumeguere Fungi

CHECK LIST OF SPECIES AND GENERA

343

gallici exsiccati, No. 995 grown
upon moist bread in Toulouse,
1880. Some member of the A.
niger group 227

A. fusco-cinereus Ellis and Morgan,
M.S. name attached to Morgan's
specimen No. 674. Evidently a
member of the .4. clavatus group
but not since seen 98

E.fuscum Pruss, in Linnaea 25: 78.
1852. Some unidentifiable mem-
ber of the .4. glaucus group.

A. fuscus Anions, in Archief voor de
Suikerindustriein Nederlandsch-
Indie 29, Deel 1, January-June
pp. 8-10. 1921. Not A. fuscus
of Bonorden, Bainier, or Schie-
mann. See: A. terreus group.. 200

A. fuscus Bonorden, in Bot. Ztg.
Jahrg. 19:202. 1861. Not iden-
tifiable 224

A. fuscus Schiemann, in Ztschr.
Induk. Abstam. u. Vererbungs-
lehre. Bd. 8(1/2): 1-35, 16 figs.,

2 pi. (1 col.). 1912 224

Syn. A. schiemanni (Schiemann)
Thorn q.v 224

A. galactus Link, cited by Yonemoto
and Kato, in Bull. Miyazaki
Coll. Agr. Forestry 3: 59-66.
1931. Perithecia formed be-
tween 15° and 28°C.

A. galeritus Blochwitz, in Ann. My-
col. 27(3/4): 205, Taf. III. 1929.
Syn. of A. terreus Thom 197

A. gallomyces Calmette, Alb. Kaiser-
liches Patent Amt. Klasse 12 q.
N. 129164 August 12 (1900). Ap-
parently a variant of A. niger.

A giganteus (Mattlet) Dodge, in

Dodge Med. Myc, p. 629. 1935. 225
Syn. S. gigantea Mattlet, in Ann.
Soc. Beige. Med. Trop 6: 35.
1926. Not A. giganteus Wehmer.
Some member of the A. niger
group.

A. giganteus Wehmer, in Centralb. f."
Bakt. 2 abt., 18(13/15): 385.
1907. Seethe A. clavatus group. 95

A. giganto-sulphureus Saito, in Jour.
Coll. Sci. Imp. Univ. Tokyo 18:

48, pi. Ill, fig. 12a to d. 1904.
Apparently a non-ascosporic
member of the A. glaucus group.
A. gigas Speg., in Ann. Mus. Nac.
Buenos Aires Ser. 3, V. 13: 434.
1911. Probably one of the A.

tamarii series 256

A. glaber Dale, nomen nudum at-
tached by Biourge to a culture
belonging with A. conicus.
S. glauca Bainier, in Bui. Soc. Bot.
France 27: 29, PI. I, fig. 3. 1880.
Also Bui. Soc. Bot. France 28:
77. 1881. Probably belongs in

A. versicolor group 193

A. glaucoides Spring, in Bull. Acad.
Sci. Belg. 19: 560-572. 1852.

Probably A. fumigatus 151

A. glaucus Link, in Obs. p. 16. 1809.

See A. glaucus group 100

mut. alba Blochwitz, in Deut. Bot.
Gesellsch. Ber. 50: 248-256.
1932. See: A. niveo-glaucus
Thom and Raper 135

var. albida Speg., in An. Mus. Nac.
Buenos Aires T. VI: 332. 1899.135

var. minimus Hanzawa, in Jour.
Coll. Agr. Tohoku Imp. Univ.
Sapporo 4: 220. 1911.

var. oblongispurus E. & W. Field
Mus. Bot. 1: 88. 1896.

var. olivascens Saccardo, in Miche-
lia 2: 543. 1878. Distributed
by Sacc. as No. 1376 Myco. ital.

var. subolivaceus Ferraris, in Fl.
ital. Crypt. P. I, fasc. 13, p. 911.
1914.

var. repens Corda, in Icones 5: 53.
1842.

Syn. A. repens DeBary q.v 103

A. globosus Jensen, in Cornell Agr.
Exp. Sta. Bui. 315, p. 482. 1912.
The strain studied by Jensen
(No. 2705) was clearly A. versi-
color 193

A. globosus Link

Syn. Sporodinia aspergillus Schro-

ter, in Sacc. Syll. 7: 207.

A. godfrini Sartory and Roederer,

in Assoc'n. Frangaise pour

l'avancement des Sciences, 42nd

344

A MANUAL OF THE ASPERGILLI

Session, Tunis, pp. 601-603.
1913. Some one of the A. glau-
cus group 133

A. gracilis Bainier, in Bui. Soc. My-
col. France 23: 92, PI. IX, figs.
11-14. 1907. In A. restrictus

series 138

var. exiguus Bainier & Sartory, in
Bull. Soc. Mycol. France 28: 47,
pi. 2. 1912. According to the
description this variety differs in
physiological characters slightly
from A. gracilis Bainier 140

A. granulatus Mosseray, in La Cel-
lule XLIII: 249-50, pi. 3, figs. 25-
28. 1934. A member of the A.
niger group; in the Biourge Col-
lection 234

A. granulosus Raper and Thorn, in
Mycologia 36: 565-568, fig. 4.
1944. In the A. ustus group. . . . 175

A. gratioti Sartory, in Compt. Rend.
Acad. Sci. (Paris) 170: 523-524.
1920. Also in Champignons
parasites de l'homme et des
animaux, pp. 578-579. 1922.
Probably some member of the A.
fumigatus group 154

A. Greconis Dodge, in Dodge Med.
Myc, p. 634. 1935
Syn. S. aurea Greco, q.v.

A. griseus Link, in Sp. Plant. Ed. 4,
6(1): 69. 1824. Incorrectly cited
by Bonorden and Wehmer as
Link Obs. 1: 69. 1809. Name
also used by Fries and by Bonor-
den. Not identified.

A. guegueni was figured inBiourge's
monograph of the Penicillia (La
Cellule t. 33, fasc. 1, pp. 7-330.
1923) as P. guegueni Plate XX.
He afterward distributed it as A.
guegueni. In A. restrictus series 139

A. guttifer Mosseray, in La Cellule
XLIII: 235-236, pi. Ill, fig. 53-
57. 1934. In the Biourge Col-
lection 233

A. gymnosardae Yukawa, in Jour.
Coll. Agr. Tokyo I: 362, PI. 18,
figs. 1-7. 1911. This fungus
was found by Yukawa under the

name "awokabi" and is de-
scribed by him as essential to the
ripening of the tunafish prepara-
tion, "katsuobushi." The di-
mensions given are intermediate
between those of A . flavus and A .
oryzae, and closely approximate
those of A. pseudo-flavus. Al-
though we have cultures related
to these forms, we have not been
able to identify these intermedi-
ates except as members of the A.
flavus-oryzae group 266

A. hageni Hallier, in Cattaneo Mico.
Corp. Urn. p. 123, PI. 6, fig. 8.

Florentin 1892 146

Syn. Otomyces Hageni Hallier, in
Zeitschr. Parasit. 1: 195. 1869.
2: 22, 233, and 259, PI. 5. 1870.
Not identifiable.

A. halophilus Sartory, Sartory, and
Meyer, in Ann. Mycol. XXVIII:
362-363, PI. Ill, figs. 11-14.
1930. Probably some nonasco-
sporic member of the A. glaucus
group 118

A. helicophorus nomen nudum at-
tached by Thaxter to a culture
that was found to belong with A.
ustus 174

S. helva Bainier, in Bui. Soc. Bot.
France 28: 78. 1881. In A.
ochraceus group 281

A. hennebergi Blochwitz, in Ann.
Mycol. 33: 238. 1935. Regarded
by Neill as one of the A. ochra-
ceus group. See A. wentii group. 249

A. herbariorum, species name seems
to appear first as Mucor herbario-
rum in Primitiae Florae Holsa-
ticae by Fredericus Henricus
Wiggers, 1780, republished in
Facsimile edition No. 23 by W.
Junk. 1925. See A. glaucus
group 100

Eurotium herbariorum ser. minor
Mangin, in Ann. des Sci. Nat.,
Bot. (Ser. 9) 10: 365. 1909. See
A . mangini 127

Eurotium herbariorum ser. major
Mangin, in Ann. Sci. Nat. Bot.

CHECK LIST OF SPECIES AND GENERA

345

(Ser. 9) 10: 365. 1909. A.glau-
cus group characterized by large
ascospores. Would fall within

A. echinulatus q.v 131

A. heterocephalus Spring, in Bull.
Acad. Sci. Belg. 19: 568. 1852.
This name was given to colonies
in a hen's egg which showed
small heads globose and large
heads columnar. Since no ade-
quate figure or description was
offered, it may be discarded as a
nomen nudum.
Physomyces heterosporus Harz. See

A. atro-ruber.
P. (Micro-aspergillus) Hickeyi, fig-
ured by Biourge in La Cellule
XXXIII, fasc. 1: 1-331. 1923
(Penicillium Monograph) , proved
to be indistinguishable from

A . gracilis q.v 139

A. hispidulus Sprengel, in Sys. Veg.
Ed. 16, 4: 541. 1827. There is
no suggestion of an Aspergillus
in the description given.
A. holmiensis Biourge, nomen nu-
dum, listed among the A. versi-
color group in Manuscript and in
culture in his collection in 1939.
A. hortai (Langeron) Dodge, in
Dodge Med. My col., p. 628.

1935 204

Syn. S. hortai Langeron, in Bui.
Soc. Path. Exotique 15: 383-384,

figs. 1-3. 1922 204

Syn. A. terreus Thorn 195

A. humicola Chaudhuri and Sachar,
in Ann. Mycol. 32: 97. 1934.
Cited by Neill as A. versicolor. . 193
A. humus Abbott, in la. St. Coll. J.
Sci. 1: 15-36. Fig. 2a-e. 1926.
See also Louisiana Station Bui.
194. One of the A. ustus series. 174
A. hypojanthinus Biourge, was pub-
lished as Penicillium hypojanthi-
num in Biourge Monogr. La Cel-
lule 33: fasc. 1, pi 321-2, PI.
XXII, fig. 130. 1923. After-
ward he transferred it to Asper-
gillus. A. restrictus series... 139

A. incrassatus Spring, in Bull. Acad.
Roy. Belg. 19: 559, fig. 2. 1852.
Not identifiable.

E. insigne Winter (exsiccati in Rabh.
Fungi No. 1732), in Hedwigia
1873 and Rabh. Krypt. Fl.
Aufl. 1, Abt. 2, 2: 61. 1887.
Probably not an Aspergillus.

S. insueta Bainier, in Bull. Soc. My-
col. France 24: 85-87, Tab. VIII,
figs. 1-13. 1908. A. ustus series. 174

A. intermedia Speg., in Myc. Arg. V.
in An. Mus. Nac. Buenos Aires
Ser. 3, T. 13: 435. 1911. In the
A. niger group.

A. itaconicus Kinoshita, in Botan.
Mag. Tokyo 45: 60-61. 1931.
Species diagnosis and figure re-
printed in Acta Phytochim.
(Tokyo) 5(3): 271-287. 1931.
See A. glaucus group 142

S. italica Sacc, in F. italici No. 109.
1881. Also in Michelia 1: 91.
1877.
Syn. A. sterigmatophorus Sacc, in
Atti. d. Soc. Ven.-Tren. d. Sc.
Nat .'2, fasc. 2: 232, Table XVII,
fig. 5-8. 1873. Some member
of the A. candidus group 211

A. janus n. sp. Raper and Thorn, in
Mycologia 36: 556-561, fig. 1.

1944 187

var. brevis Raper & Thorn, in My-
cologia 36: 561-563, fig. 2. 1944. 190

A. japonicus Saito, in Bot. Mag.
Tokyo 20: 61-63. 1906. In A.

niger group 231

var. capillatus Nakazawa, Takeda,
and Suematu. Culture avail-
able in Centraalbureau. 1939.
var. or mut. grisea Blochwitz, in
Ann. Mycol. 33: 240. 1935. It
was a change of name only, A.
malvaceus Mosseray.

A.javanicus cited by Takahashi, and
Sakaguchi, in Jour. Agr. Chem.
Soc. Japan 1: No. 10, 1925, only
in parenthesis after A . fumaricus
Wehmer and apparently deemed
asynonyn. In A. niger group.

346

A MANUAL OF THE ASPERGILLI

A. jeanselmei Ota, in Ann. de Para-
sitologic 1(2): 137-146. 1923.
A . flavus group 269

A. keratitis Ball, in Amer. Med. 2:
31. 1901. This organism was
found in an ulcer in the human
cornea. Not identifiable 147

A. koningi Oudemans, in Arch.
Neerl. Ser. II, V. 7: 284, Tab.
XIV. 1902. Not since identi-
fied.

A. laneus Link Obs. p. 16. 1809.
Syn. Botrytis lanea (Bonorden)
Sacc, in Syll. 4: 74.

A. la?ieus in Schweinitz in Syn. Am.
Bor. is syn. for Rhinotrichum
curtissii Berk, in Grevillea 3:
108. 1875.

A. laokiashanensis Shih, in Lingnan
Sci. Jour. 15(3): 368. 1933.
Near A. unguis in the A. nidu-
lans group 169

E. lateritium Montagne, in Century
VI. No. 35, Ann. Sci. Nat. Bot.
3 Ser. XI: 154. 1849. Sylloge
p. 257. One of the A. glaucus
group with ascospores 7 to 10m
in diameter.

A. Lepidophyton Wehmer.

Syn. Epidermophyton concentricum
fide Dodge, Med. Myc. 490.
1935.

A . lignier'esi Cost . and Lucet , in Ann.
Sci. Nat. IX. Bot. 2: 137, PI. 5,
figs. 19-23. 1905. This culture
from the lung of a penguin differs
in cultural details from typical
A. fumigatus especially by the
presence of swollen groups of
cells in the mycelium 159

A. longobasidia Bainier, nomen nu-
dum; a culture so labeled (Thorn
4640.477) was received from the
Bainier Collection through da-
Fonseca. This was the type of
A. bainieri Mosseray, in Ann.
Soc. Sc. Bruxelles 54, ser. B, p.
72. 1934. Member of the A.
niger group characterized by
very long primary sterigmata. . 253

A. lova,7iiensis Biourge, nomen nu-
dum; culture distributed by
Biourge and appearing in NRRL
Collection as No. 76. See Thorn
and Raper U. S. D. A. Misc.
Publ. 426, p. 18. 1941 117

A. luchue?isis Inui, in Jour. Col. Sci.
Imp. Univ. Tokyo 15: 469, PI. 22,

figs. 1-8. 1901 230

var. rubeolus Shih, in Lingnan Sci.
Jour. 15(3): 374. 1933 230

S. lutea Bainier, in Bui. Soc. Bot.
France 27: 27. 1880. A. flavus
group. See also Sartory and
Jourde, in Compt. Rend. Acad.
Sci. (Paris) 146:548-549. 1908. . 269

A. luteus (van Tieghem) Dodge, in

Dodge Med. Myc, p. 625. 1935.

Syn. S. lutea van Tieghem, in Bui.

Soc. Bot. France 24: 103. 1877.

A. luteo-niger (Liitz) Thorn and
Church, in The Aspergilli, p. 166.

1926 226

Syn. S. luteo-nigra Lutz, in Bui.
Soc. Bot. France 53: 48-52.
1907. Collected by A. Chevalier
at San Thome, Africa, in fer-
menting seeds of Theobroma
cacao. In the ^4. niger group . 226

A. luteo-niger van Luijk, nomen nu-
dum, in Biourge 's MS. p. 15.
Probably attached to a culture
of some black Aspergillus.

A. luteo-virescens Blochwitz, in Ann.
Mycol. 31(1/2): 73-83. 1932.
In A . tamarii series 256

A. lutescens Bainier, nomen nudum,
described by Thorn and Church
in The Aspergilli, p. 193, 1926.
In A. tamarii series 251

A. lutricolor Biourge, nomen nudum,
listed in MS. among the A.
ochraceus group; in the Biourge
Collection.

A. Macfiei Dodge, in Dodge Med.

* Mycol. p. 269. 1935.

Syn. Stcrigmatocystis sp. Macfie,

in Ann. Trop. Med. Parasitol.

15:279-281. 1921. Pathogenic?

In the A. niger group 239

CHECK LIST OF SPECIES AND GENERA

347

A. macrosporus Bonorden, in Hand-
buch d. allg. Myk. 1851, fig. 193.
Not identifiable.

A. malignus Lindt, in Arch. Exp.
Path. Pharm. 25: 256-271, figs.
1-11. 1889. In A. fumigatus
group 153

A. malvaceus Mosseray, in La Cellule
XLIII: 265-266, PI. 4, figs. 134-
135. 1934. A member of the A.
niger group. In the Biourge
Collection 235

A. mangini

Syn. A . minor (Mangin) Thorn and
Raper, in U. S. D. A. Misc. Publ.
426, p. 27. 1941.
Syn. E. herbariorum ser. minor
Mangin, in Ann. Sci. Nat. Bot.
(Ser. 9)10:365. 1909.
Member of the A. glaucus group
with ascospores of intermediate
size 127

A. maximus Link, in Obs., p. 16.
1809. Not an Aspergillus.

A. maydis Quevedo, in De Agronom-
ica Nos. 8 and 9, Buenos Aires,
1912. See also Sartory, in
Champignons parasites de
l'homme et des animaux p. 532,
1922. A species probably be-
longing to the A. glaucus group
described in connection with
disease in horses 147

Emericella medias Chowdhury and
Mathur, in Ann. Mycol. 36: 61-
63. 1938. Probably A. varie-
color q.v 163

A. medius Meissner, in Bot. Ztg. 55:
336-344, 354-357. 1897. See
Thorn and Raper, U. S. D. A.
Misc. Publ. 426, p. 33, 1941 133

A. melleus Yukawa, in Jour. Coll.
Agr. Tokyo 1(3): 366, Taf.
XVII. 1911. In A. ochraceus
group 279

A. mencieri Sartory et Flament, in
Compt. Rend. Soc. Biol. Paris
83:114-115. 1920. See also Sar-
tory et Bailly in Les Myocoses
Pulmonaires et leur Parasites

(Paris) pp. 180-181. 1923. A.
glaucus group; not since reported. 146

A. michelii Preuss, in Linnaea 25: 76.
1852. Not recognized.

A. microcephalus Mosseray, in La
Cellule XLIII, p. 225-227, PI. 3,
fig. 42-48. 1934. Member of
the A. niger group. In the
Biourge Collection 233

A. microsporus Boke. The descrip-
tion and figures given by Cat-
taneo and Oliva in Arch. Lab.
Bot. Critt. Garovaglio 5: 123,
PI. 6, fig. 9, 1888, have been seen.
Not recognizable.

A. micro-virido-citrinus Costantin
& Lucet, in Ann. Sci. Nat. Bot.
IX(2): 158. 1905. In A. flavus
group 263

A. minimus Wehmer, in Bot. Cen-
tralb. 80: 449-461. 1899. See
also Wehmer 's Monogr. p. 79,
1899-1901. Wehmer's culture
was lost. It has not been iden-
tified since.

S. minor Bainier, in Bui. Soc. Bot.
France 27: 30. 1880. Not rec-
ognizable. Possibly A. sydowi?

A. minor (Mangin) Thorn and Raper
in U. S. Dept. Agr. Misc. Pub.
426, p. 27-29. 1941. See A.
mangini 127

A. minutus Abbott, in Louisiana Sta.
Bui. 194. 1926. Also in Iowa
St. Coll. Jour. Sci. 1: 15-36, figs.
a, b, c, and d. 1926. See A.
ustus group 174

A. miyakoensis Nakazawa, Simo,
and Watanabe, in Agr. Chem.
Soc. Japan Jour. 12(9): 963-964.
1936. See A. niger group 220

A. mollis Bainier and Sartory, in
Bui. Soc. Mycol. France XXVII :
453, PI. XVI. 1911. Not A.
mollis Berkeley. See A. glaucus
group 129

A. ?nollis Berkeley, in English Flora
V, pt. 2, p. 340. 1836. Not
identifiable.

348

A MANUAL OF THE ASPERGILLI

A. mongolicus Biourge, nomen nu-
dum. Biourge distributed un-
der this name a culture essen-
tially like A. echinulatus (q.v.).
It appears in the NRRL Collec-
tion as No. 137. See Thorn and
Raper U. S. D. A. Misc. Publ.
426, p. 33. 1941 132

A. montevidensis Talice and Mac-
Kinnon, in Soc. Biol. [Paris]
Compt. Rend. 108: 1007-1009.
1931. See Thorn and Raper U.
S. D. A. Misc. Publ. 426, p. 26.
1941. In A. glaucus group 125

A. mouthoni Biourge, nomen nudum,
listed by Biourge among his Les
"Eurotium" in unpublished
Manuscript. Probably in cul-
ture in his collection.

A. mucoroides Corda, in Icones
fungorum II: 18, fig. 76. 1837.
Probably some member of the
A. glaucus group — unidentifi-
able.

A. mucoroideus Cook, in Grevillea
XII: 9. 1883. See A. cookei
Sacc.

A. mulleri Berkeley, in Jour. Linn.
Soc. XIII: 175. 1873. Not
identifiable.

A. mutabilis Bainier et Sartory,
in Bui. Soc. Mycol. France 27:
458, pi. XVII. 1911. In A.
glaucus group 129

A. mycetomi Villabruzzi and Gelo-
nesi,in Annali Med. Nav. Colon.
33: 283-308, 8 figs. 1927.
Syn. Madurella sp.; undeter-
minable from the data avail-
able.

A. nujcobanche Link, in Sp. Plant
Ed. 4, 6: 65. 1824. A fungus
from rotting Peziza — not identi-
fiable.

A. nantae Pinoy, in Compt. Rend.
Soc. Biol. 97(19): 67-68. 1927.
This was probably A. unguis in
the A. nidulans group. Listed
by Biourge as in his collection,
1939 170

A. nanus Montagne, in Sylloge
Generum Specierumque Crypt,
p. 300, No. 1112. Paris. 1856.
Also in Saccardo Sylloge
Fungorum 4: p. 71. Patavii

1886. In the A . niger group 231

A. nanus Oudemans, in Nederl.
Kruidk. Arch. Ser. 3, 2: 1121.
1904. Not A. nanus Mont. q.v.
Probably some conidial form in
the A. glaucus group.
E. nebulosum Fries, in Sys. Myc. 3:
334. 1832. No data are given
to link this with Aspergillus.
A. nicollei Pinoy, cited by Biourge
in 1939 manuscript. He has
apparently raised to species
rank S. nidulans var. nicollei

Pinoy, q.v 170

A. nidulans (Eidam) Wint., in Rab.

Krypt.-F1.12:62. 1884 156

Syn. S. nidulans Eidam, in Cohn,
Beitr. Biol. Pfeanzen 3: 392-411,
PI. 20-22. 1883.
forme Cesarii Pinoy, in Bul. Soc.

Path.Exot.8:ll. 1915 170

mut. coerulea Blochwitz, nomen
nudum upon a culture in the-
Biourge Collection, 1939.
Listed in Biourge MS.
mut. alba E. Yuill, in Jour. Bot.
(London) 1939, p. 175, pi.

618 159

var. latus Thorn and Raper, in
Mycologia 31(6): 657-9.

1939 159

var. Nicollei Pinoy, in Compt.
Rend. Acad. Sci. Paris 144:
396. 1907. This variety was
found fruiting within human
tissue in a subject affected with
"Madura-foot." Listed in
Biourge's Collection, 1939, as

S. nicollei Pinoy 170

Syn. S. nidulans var. Nicollei
Pinoy, in Archives d. Para-
sitologic 10: 437-458, PI. XL
1906.
Diplostephanus nidulayis (Eidam)
Langeron, in Compt. Rend. Soc.
Biol. Paris 87: 343-345. 1922.

CHECK LIST OF SPECIES AND GENERA

349

Syn. for A. nidulans Eidam

q.v 156

A. niger van Tieghem, in Ann. Sci.
Nat. Bot. Ser. 5, V. 8(4): 240.
1867. See also Thorn and Cur-
rie, in Jour. Agr. Res. 7: 1-15.
1916 216

Syn. S. nigra van Tieghem, in Bui.
Soc. Bot. France 24: 102-103.
1877.

Syn. Aspergillopsis nigra (v. Tieg-
hem) Speg., in Myc. Arg. V, in
Ann. Mus. Xac. Buenos Aires
(ser. 3) 13: 435. 1911.

mut. cinnamomeus n. comb. See:
A . cinnamomeus Schiemann 223

var. laevis Blochwitz, in Ann.
Mycol. 33: 249. 1935. Pro-
posed for smooth-spored strains.
A. niger var . fermentarius Nakazawa,
Simo, and Watanabe, in Jour.
Agr. Chem. Soc. Japan 144: 171-
2 and 184. 1936 (in Japanese) ... 220

mut. fusca Blochwitz, in Ann.
Mycol. 32: 87. 1934. In the
A. niger group. Such a separa-
tion based upon conidial color is
of doubtful validity.

mut. Schiemanni n. comb 224

See: A. schiemanni (Schiemann)
Thorn.

forma Tuebingen Schober, in the
Centraalbureau, is apparently
the strain used in the researches
of Schober.
A. niger citricus Wehmer, name on a
culture received from Xeuberg
(C. T. Xo. 4668.4) but without
description.

Syn. A . citricus Mosseray q.v 235

S. nigra Bainier. Syn. for S. phoeni-
cis (Corda) Patouillard and
Delacroix, in the A. niger series.
A. nigrescens Robin, in Histoire
Xaturelle des Vegetaux Para-
sites (Paris) p. 518, pi. 5, fig. 2.
1853. Xo success has been
made in interpreting Robin's
organism 151

A. nigricans Wreden, in Compt.
Rend. Acad. Sci (Paris) 65: 368.
1867. Probably A. niger group.

A. nigriceps B. and C, in the Curtis
Collection. Specimen collected
by Charles Wright (No. 927) in
Cuba. Cited by Cooke, M. C.
in Grevillia 17: 21, Sept. 1888.
A slide from this material
prepared by Bullard and pre-
served in the Harvard collection
shows a characteristic organism
of the A. niger series, not sepa-
rable from A . niger van Tieghem.

A. niveocandidus Lindau, in Rabh.
Krypt. Fl. 8: 151. 1907. In the
A. candidus group.

A. niveo-glaucus Thorn and Raper,
in U. S. Dept. Agr. Misc. Pub.
426, p. 35-36. 1941 135

A. niveus Blochwitz, in Ann. Mycol.
27(3/4): 205-206, Taf. Ill, fig.
2. 1929. A member of the A.

terreus group 202

var. major Blochwitz, in Ann.
Mycol. 32(1/2): 86. 1934. Ap-
parently belongs in the A.

candidus group 211

var. nubila Blochwitz, in Ann.
Mycol. 32(1/2): 85. 1934. See
A. carneus 202

A. Nblting Hallier, in Zeitschr.
Parasit. (Xot found) cited
by Cattaneo and Oliva in Arch.
Lab. Bot. Critt. Garovaglio 5:
122. 1888. Xot recognizable.

A. novus, nomen nudum attributed
to Wehmer; culture in the Cen-
traalbureau at Baarn was identi-
fied by Thorn and Raper as A.
pseudo-glaucus Ill

E. obliteratum Schw. Syn. fungorum
in Amer. Bor. X. 2725. Some
unidentifiable member of A.
glaucus group.

A. oblongisporus E. and E., Xo. 760
in Xuttall's Flora of Fayette
County, West Virginia. Listed
apparently by Millspaugh in
"The Living Flora of West Vir-
ginia" in W. Va. Geol. Survey

350

A MANUAL OF THE ASPERGILLI

5(A): 32. 1913. as .4. glaucus
var. oblongisporus. E. & W.
Examination of this material
shows it to be a mixture of A.
flavus and A. repens.

S. ochracea Bainier, in Bui. Soc.
Bot. France 28: 78. 1881.
Some member of the A . ochraceus
group.

S. ochracea Delacroix, in Bui. Soc.
Mycol. France 7: 109, PI. VII,

fig. f. 1891 282

Syn. S. delacroixii Sacc, in Syi-
loge 10: 527. Delacroix de-
scribed his organism without
recognizing the previous use
of the specific name 282

S. ochracea (Wilhelm) Schroter.
See: A. ochraceus Wilhelm.

A. ochraceo -ruber Sacc, in Miche-
lia I: 77. 1877. (Saccardo, P.
A. No. 1063 in Myco. Veneta, on
bark of Walnut, 1876, see fig. 17
in Fungi italici). Some conidial
form of the A. glaucus group.

A. ochraceus Wilhelm, Inaug. Diss.

Strassburg. p. 66. 1877 279

var. microspora Tiraboschi, in
Annali di Botanica 7: 14.
1908 281

S. ochroleuca Speg., in Anal. Mus.
Nac. Buenos Aires, Ser. 3, t.
13: 434. 1911. In the A. ochra-
ceus group 276

A. ochroleucus Haller, in Enum.
Method. Stirp. Helvet. Indig. t.
I: p. 6. 1742. Hist. Stirp. t.
III. 1768. Cited by Gasperini
as questionable syn. for A.
elegans.

A. okazakii Okazaki, in Centralb. f.
Bakt. 2 abt. 42(10/14): 225.

1914. A. candidus group 211

This is cited in the Sylloge 22:
1260 as >S. okazakii (Saito) Sac-
cardo.

A. oligosporus Corda, in Icones III,
Tab. II. Not an Aspergillus.

S. olivacea van Tieghem, in Bull.
Soc. Bot, France 24: 103. 1877.
Not identified with A. olivaceus

Preuss, 1852. Not identifiable.
A. olivaceo-fuscus Mosseray, in La
Cellule XLIII: 258-259. pi. 3,
fig. 49-52. 1934. A member of
the A. niger group in the

Biourge Collection 234

Cladosarum olivaceum Yuill and
Yuill, in Trans. Brit, Myc. Soc.

22: 194-200, PI. 11-13. 1938 73

A. olivaceus Delacroix, in Bui. Soc.
Mycol. France 13: 118-120, text
fig. 1897
Syn. .4. delacroixii Sacc. and
Sydow q.v. Not A. olivaceus
Preuss. 1852. Probably in A.
glaucus group.
A. olivaceus Preuss, in Linnaea 25:

77. 1852. Not identifiable.
A. Oniki on a culture from Okunuki
in the Centraalbureau collection.
No description found. Bloch-
witz cites it in Ann. Mycol. 33:
240. 1935. As A. ochraceus.
A. oosporus Wallroth, as Flora
Cyptogamica Germaniae No.
1928, in Compendium Florae
Germanicae 4: 296. 1833.
Some one of the A. glaucus
group.
A. oriolus nomen nudum attributed
to Biourge. Distributed by
Biourge Collection and appears
as NRRL No. 87. It was iden-
tified as a strain of A . chevalieri
by Thorn and Raper in U. S. D.
A. Misc. Publ. 426, pi. 21. 1941. 120
A. oryzae (Ahlburg) Cohn, in
Jahresb. Schles. Ges fur Vaterl.
Cultur 21: 226. 1883. In the

A. flavus -oryzae group 261

Syn. E. oryzae (Ahlb.) Korschelt.
Name with incomplete descrip-
tion of sake organism published
in Dingler's Polytech. Jour.

230:330. 1878 261

A. oryzae var. basidiferens Constan-
ts and Lucet, in Ann. Sci. Nat,
Bot. IX (2): 167. 1905. In
the A. flavus-oryzae group.
Syn. A. oryzae var. basidifer Sacc.
Syll. 22.

CHECK LIST OF SPECIES AND GENERA

351

A. ostianus Wehmer, in Bot.
Centralb. 80: 449 461. 1899.
Monogr. pp. 117-119, Taf. II.
No. I. 1899-1901. In the A.
ochraceus group 283

A. oval is perm us Link, in Obs. II, p.
37, 1816; also in Sp. Plant Ed. IV,
Vol. VI(1): 66. 1824. Cited as
synonym of A. oosporus Wallr.
(1833) without evidence of iden-
tity or reason for redescription.

A. panamensis Raper and Thorn, in
Mycologia 36: 568-572, fig. 5.
1944 242

A. parasiticus Speare, in Hawaiian
Sugar Planters Exp. Sta., Path.
& Physiol. Ser. Bui. 12: p. 38,
PI. 3 and 4. 1912. In the A.

flavus group 266

Syn. A. flavus var. viridis Bloch-
witz, q.v 267

A. penicillatus Greville, in Scottish
Cryptogamic Flora 1: 32, plate
32. 1823.
Syn. A. glaucus (conidial); not
identifiable to species.

A. penicillatus Link, in Sp. Plant
Ed. 4, 6(1): 69. 1824. The
description given by Link is not
sufficient to identify any form,
but since he probably intended
to cover the organism of Gre-
ville, it may be assumed to be
conidial A . glaucus. Sturm cites
it as Briarea elegans without
specifying his reasons.
A. penicilloides Spegazzini, in Rev.
Agrar. Veter. La Plata, p. 245.
1896. In the A. restrictus

series 142

A. penicillopsis (Hennings) Racib.
P. Hennings as Stilbothamnium
penicillopsis P. Henn. & E.
Nym. described in Fungi Mon-
sunensis (Warburg-Monsunia,
Bd. I: p. 37 (Leipzig). 1899.
Exsiccati of type in Pathological
collections U.S. Dept. Agr. Bur.
of Plant Industry as : Raciborski
No. 87, in Crypt. Paras. Java.

See also Paras. Alg. u Pelz. Javas

II: 7. 1900 282

A. periconioides Sacc, in Ann.
Mycol. 11: 320. 1913. No. 195
in Sydow Fungi exotici exsic-
cati collected by P. W. Graff on
leaves of Carica in Luzon, 1912.
Not identifiable.
A. pemiciosus Inui., in Jour. Coll.
Sci. Imp. Univ. Tokyo 15: 473.
1901. Inui recorded a yellowish
greenish color in the mycelium of
this species which is regarded as
related to A. luchuensis and A.

niger 230

A. pertardus is a nomen nudum on a
culture distributed by Biourge.
The organism was close to A.

restrictus series 142

A. petiolatus Haller, in Historia stir-
pum indigenarum Helvetiae
inchoata, etc. 1768.
A. phaeocephalus Durieu and
Montagne, in Fl. Alg. p. 342.
1849.
Syn. S. phaeocephala (Dur. et
Mont.) Saccardo, Fungi italici
fig. 903 and No. 1244 in Sacc.
Myc.Veneta. 1877. One of the
A. niger group.
A., phoenicis (Corda) Thorn, in The

Aspergilli,p.l75. 1926 222

Syn. S. phoenicis (Corda) Patouill.
and Delacr., in Bui. Soc. Mycol.

France 7: 119, PI. 9. 1891 223

Syn. Ustilago phoenicis Corda, in
Icones Fungorum 4: 9, pi. 3,
fig. 26. 1840. One of the A.

niger group 223

A. pictor Blanchard, cited by Castel-
lani & Chalmers in Man. Trop.
Med. p. 806. 1913.
Syn. Trichophyton pictor Blan-
chard, in Traite Path. Gen. II:
919. 1896. The production of
polychromatic cultures suggests
relationship to A. versicolor.
A. pollinis Howard, in Am. Bee
Jour. 36: 577-578. 1896. See
also idem. 38: 530-531. 1898.
This was A. flavus fide Turesson

352

A MANUAL OF THE ASPERGILLI

in Svensk Bot. Tidskr. 11: 30.
1917 266

S. polychroma Ferraris, in Fl. It.

Crypt. Hyph. p. 640. 1906 193

Syn. A . versicolor q.v 190

A. polychromus De Mello, in Jour.
Indian. Bot. 1(5): 158-161.
1920. The data given are not
sufficient to distinguish whether
he had A. nidulans or A.
sydowi.

A. polychromus Sartory, Sartory,
and Meyer. Cit. Syn. of A.
versicolor fide Blochwitz in Ann.
Mycol. 31: 73. 1933.

A. polymorphous Moquin-Tandon, in
Elements Bot. Med. 2 Ed., 469.
1866. Not identifiable.

A. pouchetii Montagne, in Ann. Sci.
Nat. Bot. 4 Ser. 6(12): 182-183.
1859. As described this was one
of the mucors, noted as having
resemblances to A. maximus
(Sporodinia).

A. praecox Mosseray, in La Cellule
XLIII: 229. 1934. Cited as
synonym of A. fuliginosus
Peck 233

*S. prasina Bainier, in Bull. Soc.
Bot. France 27: 31. 1880. Not
identifiable.

A. profusus Hann, nomen nudum.
Cited by Thorn and Raper in
discussing A. scheelei Bainier
and Sartory (Bui. Soc. Myc.
France 28: 257. 1912) as nearly
related to A . scheelei. See Thorn
and Raper, U. S. D. A. Misc.
Publ. 426, p. 13, 1941. Syn. of
A. pseudoglaucus Ill

A. proliferans Geo. Smith, in Brit.
Mycol. Soc. Trans. 26(1/2): 26,
PI. III. 1943. In the A.
ruber section of the A. glaucus
group 117

A. pseudo -carbonari us (Bainier)
Mosseray, in La Cellule XLIII:
224-225, PI. 3, fig. 7-13. 1934. . . 233
Syn. S. pseudo-carbonaria nomen
nudum on culture (Thorn 4640.-
482) from the Bainier collection.

A. pseudo-citricus Mosseray, in La
Cellule XLIII: 228-229, pi. 4,
figs. 103-104. 1934. Member
of the A. niger group; in the

Biourge Collection 233

A. pseudo-clavatus Purjewicz, in
Schrift. Naturforch. Gesell.
Kiev. 16, 2, p. 309, 1900. See:
Saccardo Sylloge Fung. 16, p.

1028. In A. clavat us group 98

A. pseudo-elatior Mosseray, in La
Cellule XLIII: 255-256, pi. 3,
figs. 33-37. 1937. Member of
the -4. niger group; in the

Biourge Collection 1939 234

A. pseudoflavus Saito, in Centralb.
f. Bakt. 2 abt. 18(1/3): 34, figs

15-18. 1907 266

Syn. S. pseudoflava Sacc. Sylloge
Fungorum 22: 1260-1266. The
morphology given indicates that
A. pseudoflavus is one of the
intermediate forms which bridge
the gap between typical A . flavus
and A. oryzae.
A. pseudoglaucus Blochwitz, in Ann.
Mycol. 27: 207. 1929. Emended
description by Thorn and Raper
in U. S. D. A. Misc. Publ. 426,
p. 12, 1941. A. glaucus group.. . . 110
S. pseudo -nidulans Vuillemin, Arch.
Parasitologie 8: 540-542. 1904.
Vuillemin transfers the asco-
sporic form described by Grijns
as A. fumigatus in Centralbl.
Bakt. II, 11: 330. 1903, to this
specific name, emending Grijns's
description by indicating the
double nature of the band by
which he separates his form from
A. ?iidulans as described by
Eidam. This discussion by
Vuillemin tallies with the com-
monest of our American soil
forms of A. nidulans but not
with the description by Grijns.
A. pseudo-niger Mosseray, in La
Cellule XLIII: 256-258, PI. 4,
figs. 113-117. 1934. A member
of the A. niger group; in the
Biourge Collection. 1939 234

CHECK LIST OF SPECIES AND GENERA

353

S. pseudo-nigra Costantin and Lucet,
in Bui. Soc. Mycol. France 19:
33-44. 1903. In the A. niger
group.
A. -pseudo -Schiemanni Biourge,
nomen nudum, cited from Cen-
traalbureau catalogue 1931, as
an actively diastatic organism
represented in the Biourge Col-
lection.

A. pulchellus (Speg.) Thorn and
Church, in The Aspergilli, p. 181.

1926 229

Syn. Aspergillopsis pulchella
Speg., in Myc. Arg. V, An.
Mus. Nac. Buenos Aires, Ser. 3,
T. 13: 436. 1911. In the A.
niger group 229

E. pulcherrimum Winter, in Rabh.
Krypt. Fl. 2, aufl. 1 Abt. 2:
60. 1887. Noted there as also
in Herbarium of Winter and in
Hansen, Fungi fimicoli Danici
1041 of Sep. Abdr. A copro-
philus form from the dung of
foxes in Leipzig and dogs in
Denmark, by Hansen; not an
Aspergillus.

A. pulmonum hominis Welcker.
Discussed by von Dusch, in
Virchow's Archiv. (N. F. 1) 11:
561-566. 1857. Apparently
A.fumigatus 151

A. pulverulentus (McAlpine) Thorn,
in Jour. Agr. Res. 7: 10-11.

1916 223

Syn. S. pulverulenta McAlpine, in
Agr. Gaz. N. S. Wales 7: 302.
1896. In the A. niger group 223

A. pulvinatus B. and C. original
collection as far as seen appears
to be in the Curtis Collection
from Society Hill, S. C. 1855;
also one marked F. cub. — Wright
No. 642 in the Curtis Collection;
another series of specimens of B.
and C. No. 1648 in Ellis and
Everhart N. A. Fungi collected
on dead twigs at Newfield, N.J.
1885. Also No. 2306 in the Ellis
Collection. Not an Aspergillus.

*S. purpurea van Tieghem in Bui.
Soc. Bot. France 24: 101-103.
1877. Possibly A. nidulans.
A. purpureofuscus Fries, in Sys.
Myc. 3: 388. 1829. Probably
the same as A. purpureofuscus
of Schweinitz.
A. purpureofuscus Schweinitz, in
Synopsis fungorum in America
boreali media degentium.
Secundum observationes. In
Trans. Amer. Phil. Soc, N. S.
4: 282, No. 2680. 1834. Also in
Saccardo Sylloge Fungorum 4:
68, Patavii. 1886. Not an
Aspergillus.
A. purpureus Haller. Historia
stirpum indigenarum Helvetiae
inchoata, etc. 1768. Not
recognizable.
S. pusilla Peyronel, in I genu atmos-
pherici dei funghi con micelio.
Thesis. Padova p. 21. 1914.

See : the A . niveus series 203

A. pusillus Massee, in Kew. Bull.
Misc. Inf. 4: 158. 1914. From
Soil, Sudan. Not identifiable.
A. pyri English, nomen nudum,
name published in Research
Studies of the State College of
of Washington VIII (3): 127.
1940. (Doctoral Thesis: Taxo-
nomic and pathogenicity studies
of the fungi which cause decay
of pears in Washington.) Sub-
sequent work by English led to
recognition of the name as a

synonym of A . niger 231

A. quadrifidus Link, in Obs. 2: 36.
1816. Probably not an Asper-
gillus.
A. quadrilineatus Thorn and Raper,
in Mycologia 31(6): 660, fig. 3D

and4B. 1939 160

A. quercinus (Bainier) Thorn and
Church, in The Aspergilli, p.

186-187. 1926 276

Syn. S. quercina Bainier, in Bui.
Soc. Bot. France 28: 78.
1881 276

354

A MANUAL OF THE ASPERGILLI

A. quininae Heim., in Bui. Soc.
My c. France 9: 239. 1894. The
culture was found upon quinine
solution but the description
given will not separate it from
A. fumigatus.

A. racemosus Persoon, in Neues
Mag. Bot. 1: 121. 1794. Also
in Tentamen Disp. Meth. Fung,
p. 41. 1797. Not recognizable.

A. ramosus Hallier, in Ztschr. f.
Parasit. 2: 266-269, PI. 6, figs.
1-6. 1870. The figures and
descriptions evidently represent
a strain of A. fumigatus 151

A. raulini, nomen nudum, in
Biourge's table, probably at-
tached to a culture in his collec-
tion.

A. rehmii Zukal, in Oesterr. Bot.
Zeitschr. 43: 160, PI. II, figs.
1-10. 1893. Zukal regarded
this form as close to S. sulphurea
Fresenius, but his description of
perithecia and ascospores ex-
cludes the A. ochraceus group.
Blochwitz evidently believed
that Zukal had a mixed culture;
hence, the name would be unten-
able. Cultures belonging to the
A. ochraceus group have been
distributed under the name but
without proving their authority 281

A. repandus Bainier and Sartory, in
Bui. Soc. Myc. France 27: 463,
Pi. XVIII. 1911. A. glaucus

group.

133

A. repens (Cda.) DeBary, in Ab-

handl. I. Senkenberg. Naturf.

Gesellsch. 7: 379. 1870 103

A. repens DeBary and Woronin, in

Beitrage zur Morphologie und

Physiologie der Pilzen p. 379.

1866.
Syn. A. glaucus var. repens Corda,

in Icones 5: 53, Taf. II, fig. 24.

1842 103

E. repens var. amstelodami Vuill.,

Soc. Mycol. de France, Bui.

Trimest.36:131. 1920 122

A. restrictus G. Smith, in Jour.
Text. Inst. 22: T. 115, fig. 5.

1931 141

A. restrictus var. B., G. Smith, in
Jour. Text Inst. 22: T. 115, figs.
4, 6, and 8. 1931. See A.
restrictus series in A. glaucus

group 141

A. roseus Batsch, in Elenchus Fung-
orum, p. 183, No. 58, fig. 58.
. 1783. Cited by Link, Spec. PL,
ed. IV, t. VI, pt. 1, p. 68. 1824.
Also by various authors for a
rosy or flesh-colored organism,
and by Corda as Haplotrichum
roseutn in Pracht-flora, PI. XI.
A. roseus Link, in Sp. Plant, ed. IV,
t. 6, part 1, p. 68. 1824. Link
took the name "roseus" used
descriptively, but not nomen-
clatorially, by Batsch (El. Fung,
p. 183, no. 58, fig. 58. 1783) for
a mold presumed to have been
an Aspergillus, by later authors.
The name appears as No. 2724 in
the Curtis Collection (1849) for a
member of the A. candidus
group. Neither descriptions
nor specimen dating back to
these authors fix this name for
any definite series.
A. rubens Green, in Boston Soc. of
Med. Sc. 1868. Not identifi-
able.

A. ruber (Bremer) 114

Syn. A. ruber (Spieckermann and
Bremer) Thorn and Church, in

The Aspergillill2. 1926 114

Syn. Eurotiurn rubrum Bremer, in
Zeitschr. f. Untersuch. d. Nah-
rung. und Genussmittel IV. 1901,
p. 72; also in Die fettverzehr.
Organismen in Nahr. u. Futter-
mitteln, " Dissert. Munster.

1902 114

Syn. E. rubrum Spieckermann and
Bremer, in Landw. Jahrb. 31:

81-128. 1902 114

A. ruber Estienne

Syn. Physomyces heterosporus
Harz.

CHECK LIST OF SPECIES AND GENERA

355

Syn. S. rubra (Estienne) Biourge.
Syn. Monascus pur pit re us; cer-
tainly not an Aspergillus.
S. rubescens nomen nudum on a cul-
ture in the Bainier Collection
(Thorn No. 4640.487). A.

flavipes group 181

A. rufescens Berlese, in Fungi Mori-
colae Fasc. VII. No. 4, Tav. 54,
figs. 12-17. 1889. Probably
conidial strain of .4. glaucus
group.
A. rugulosus Thorn and Raper, in
Mycologia 31(6): 661-2; fig. 3E

and4C. 1939 160

A. rutilans Mosseray, in La Cellule
XLIII. 234-235, PI. 4, fig. 86-
90. 1934. A member of the A.
niger group; in the Biourge

Collection 1939 233

E. sacchari Spegazzini, in Anales del
Museo Nacional de Buenos
Aires, 6, Ser. 2, 3: 244. 1899.
Some insufficiently described
member of the A. glaucus
group.
A. sachari Chaudhuri and Sachar, in
Ann. Mycol. 32: 95. 1934.
Blochwitz in Ann. Mycol. 33:
240, 1935, leaves this species in

A . quercinus 278

A. salmoneus Biourge, nomen
nudum, cited by Henrard in La
Cellule XLIII: fasc. 2, p. 353.
1934, as one of the series
"glauci." On p. 370, idem, he
records that single ascospores re-
quired 3 months for germination
and that this species was found
to be homothallic. Identifica-
tion to species within the A.
glaucus group is not possible
without more information.
Alliospora Sapucaya Pirn, in Proc. R.
I. Acad. 1883 and in Jour. Bot.
1883. p. 234. One of the A.
niger group causing rot in allia-
ceous bulbs 9

A. sartoryi nomen nudum was
attached by Biourge to a culture
of an organism close to A.

gracilis in the A. rcstrictus

series 139

A. sartoryi Sydow, in Ann. Mycol.
11: 156-160, PI. VIII. 1913.
Possibly in .4. tamarii group.

A. scheelei Bainier and Sartory, in
Bui. Soc. Myc. France 28: 257-
262, PI. X. 1912. See Thorn
and Raper in U. S. D. A. Misc.
Publ. No. 426, p. 12. 1941. A.
glaucus group 107

A. scheelei Bainier and Sartory var.
B., idem. See A. repens series
in A. glaucus group 107

A. schiemanni (Schiemann) Thorn, in
Jour. Agr. Res. 7: 13. 1916.
See also A. fuscus Schiemann;
name changed because previ-
ously used 224

S. schneggiana Biourge, nomen
nudum, in Biourge 's MS; listed
in the A. candidus group as
applying to a culture received
from Schnegg.

A. sclerotifer Mosseray, in La Cellule
XLIII (fasc. 2): 247-248. 1934.
A member of the A. niger

group.

233

A. sclerotiorum Huber, in Phyto-
pathology 23(3): 306-8, fig. 1.
1933. A heavy sclerotium-
producing member of the A.
ochraceus group 278

A. sejunctus Bainier et Sartory,
in Bui. Soc. Myc. France 27:
346-368, Pis. X-XI. 1911. An
ascosporic form of the A . glaucus

group.

114

E. semi-ituniersum Marchal, in C. R.
Soc. Roy. Bot. Belg. 33: 128, Pi.
II, fig. 3. 1895. Not an Asper-
gillus.

A. siebenmanni Costantin and Lucet,
in Ann. Sci. Nat. Bot. 9, 2, p.
162. 1905. This name is based
upon Siebenmann's description
of an organism from the human
ear identified by Siebenmann as
A. flarus, but regarded by the
describers as a separate species

356

A MANUAL OF THE ASPERGILLI

based upon the description
given by Siebenmann 266

A. simplex Persoon, in Tent. Disp.
Meth. Fung. p. 41. 1797.
Tradition calls it a Penicillium.

A. soya on a culture from Okunuki in
the Centraalbureau collection.
No description found; cited by
Blochwitz in Ann. Mycol. 33:
240. 1935, as A. flavus.

S. skottsbergii Bresadola and Vester-
gren no. 250 in Vestergren,
Micromycetes rariores selecti
Rossia baltica: ins. Osilia, Kiel-
kond in silva abiegna prope
Kattiel in foliis vivis Aqulegiae
vulgaris. 1899. Distributed
without description; examined in
collaboration with Professor
Thaxter at the Harvard Uni-
versity Cryptogamic Her-
barium; did not prove to be an
Aspergillus.

A. spadix Amons, in Arch. v. Suiker-
industrie in Nederlandsch Indie
29: 12-14. 1921. This is one of
the A . tamarii series 257

A. sparsus Raper and Thorn, in
Mycologia 36: 572-574, fig. 6.
1944 283

A. sphaerospermus Corda, in Icones
II: 18. 1854. No description.

A. spiralis Grove, in Journal of Bot.
23: 164, tab. 257, fig. 5. 1885.
A conidial organism of the A.
glaucus group.

A. spirius cited by Amons in Arch,
v. d. Suikerindustrie in Neder-
landsch-Indie 29: 14. 1921.
Not identifiable.

S. spuria Schroeter, in Cohn,
Kryptogamen Flora von Schles-
ien 3: 2 Halfte, Lief. 1, p. 218.

1893 201

Syn. ? S. carnea van Tieghem.
1877. See: A. cameus (van
Tieghem) Blochwitz 201

A. stellatus Curzi, in Rend. Acad.
Naz. Lincei 19: 424-428. fig. 1.
1934. Culture in Centraal-
bureau list 1939.
Syn. A.variecolor 163

E. stercoraria Hansen, in Meddelelser
fra den Naturhist. Foren. i.
Kjobenhavn p. 310. 1876.
Syn. Anixiopsis stercoraria
Hansen, in Bot. Ztg. 55: 127-131,
Tab. II, fig. 8. 1897.

A. stercoreus Sacc, in Michelia 1: 78.
1877. Also Fungi italici no. 19.
An unidentifiable member of the
A. glaucus group.

A. sterigmatophorus Saccardo, in Atti
Soc. Ven.-Tren. Sci. Nat. 2,
fasc. 2: 232, Tab. XVII, fig. 5-8.

1873 211

Syn. S. italica q.v 211

A. sirychni Lindau, in Hedwigia
Bd. 43, Heft 5: 306-307. 1904.... 223
Syn. A. pulverulentus McAlpine,
1896. In the A. niger group 223

A. subfuscus Johan-Olsen, in Medde-
lelser fra Naturh. forening i
Kristiania. 1885. Cited also
in O. Johan-Olsen, Viden-
skapselkabet i Kristiania p. 21.
1886. Some member of the A.
niger group 231

A. subgriseus Peck, in Bui. Torrey
Bot. Club, 22, 5: 210. 1895.
Syn. E. subgriseum Peck, in Rept.
N. Y. State Mus. Bot. p. 30.
1910. Also in N. Y. State Mus.
Bui. 150. 1911. There is no
way to identify Peck's species.

A. sulphureus (Fres.) Thorn and
Church, in The Aspergilli, p.

185-186. 1926 275

Syn. S. sulphurea Fresenius, in
Beitr.Z.Mykologie,Heft3:p.83,
Tab. XI, fig. 30-33. 1863. Also
Sylloge 4: 73. 1886. In A.
ochraceus group 275

A. sydoivi (Bainier and Sartory)
Thorn and Church, in The Asper-
gilli, p. 147-148. 1926 184

Syn. S. sydowi, Bainier et Sartory,
in Ann. Mycol. 11: 25-29, PI. III.

1903. /I . sydowi series 184

var. achlamydosporus Nakazawa,
Simo, and Watanabe, in Jour.
Agr. Chem. Soc. Japan 10(2):
178-179. 1934. The absence of

CHECK LIST OF SPECIES AND GENERA

357

Hulle cells is inadequate for
separation 186

A. syncephalis Gueguen, in Champ.
Parasit. Horn. Anim. p. 165, fig.
6. 1904. Probably A. fumi-
gatus 151

S. szurakiana Moesz, in Bot. Kozlem.
19: 44-66, 13 figs. 1921. One
of the A. candidus group.

A. tabacinus Nakazawa, Simo, and
Watanabe, in Jour. Agr. Chem.
Soc. Japan 10(2): 177-178.
1934. Appears to have been a
strain of the A. versicolor
series 193

A. tamarii Kita, in Centralb. f.
Bakt. etc., 2 Abt. 37, No. 14/16,
pp. 432-452. 1913. A tamarii
series 254

A. tardior Biourge, nomen nudum,
listed among his cultures of the
A. restrictus group (Biourge 's
Sec. I. Microaspergilli).

A. terreus Thom, in Turesson, Gote,
Svensk Botanisk Tidskrift 10:
5. 1916. Without description;
diagnosis Thom and Church
Amer. Jour. Bot. 5: 85-86. 1918.

The A . terreus series 195

var. aureus, n. var 198

var. Boedijni (Bloch.) n. comb.

Thom and Raper.
Syn. A. Boedijni Blochwitz, Ann.

Mycol. 32(1/2): 83. 1934 197

var. floccosus Shih, in Lingnan
Science Jour. 15: 372, pi. 16, fig.

3. 1936 198

var. subfloccosus Shih, in Lingnan

Science Jour. 15: 371, pi. 16, fig.

4. 1936.

A. terricola Marchal, in Rev. Mycol.
15(59): 101-103. 1893. In A.

tamarii series 253

var. Americana Marchal in Thom
and Church, Am. Jour. Bot. 8:
125. 1921. Also in Thom and
Church The Aspergilli, p. 192.
1926 253

A. thoynii. This was an undescribed
sclerotium-producing strain of
A. flavus sent by P. W. Graff to

Amer. Type Cult. Collec-
tion 266

A. tiraboschii Carbone, in Atti d.
Inst. Bot. Univ. Pavia Ser. II
Vol. XIV: 320. 1912. In the
A. sydowi -versicolor group 188

A. tokelau Wehmer, in Centralbl.
Bakt. I, 35: 140. 1903. Cited by
Dubreuihl in Jour. Med. Bor-
deaux 32: 312. 1902. Wehmer's
culture was one of the A . glaucus
group but not the pathogenic
organism causing the disease
"tokelau." Dodge, Med Myc.
490. 1935. calls it Epider-
mophyton.

S. tropicalis Matta, in Bol. Inst.
Brasil. Sci. 3: 51-54, 2 figs.
1927. Probably A. sydowi but
not separable from other strains.

A.tubingensis (Schober) Mosseray, in
La Cellule XLIII: 245-247, pi.
3, fig. 58-60. 1934. Member
of the A. niger group; in the
Biourge Collection 1939 234

A. tunetanus (Langeron) Dodge, in
Dodge Med. Mycol., p. 635.
1935.
Syn. S. tunetana Langeron, in Bui.
Soc. Path. Exot. XVII: 345-347,
illust. 1924. See A. sydowi
series 186

A. umbrinus Patterson, in Bull.
Torrey Bot, Club 25: 284. 1900.
In the A. tamarii series.

A. umbrosus Bainier and Sartory, in
Bui. Soc. Mycol. France 28: 267,
PI. XII. 1912. In the A.
glaucus group 129

A. unguis (Emile-Weil and Gaudin)
Emend. Thom and Raper in
Mycologia 31(6): 667-8, fig. 6.

1939 169

Syn. S. unguis Emile-Weil and
Gaudin, in Arch. Med. Exp.
Anat. Path. (Paris) 28: 463-465.
1919. See Thom and Raper,
The A. nidulans group, Mycolo-
gia 31: 667. 1939 169

A. ustus (Bainier) Thom and Church,
in The Aspergilli, p. 152-153.
1926 171

358

A MANUAL OF THE ASPERGILLI

Syn. S. usta Bainier, in Bui. Soc.
Bot, France 28: 78. 1881. .4.

iistus group 171

var. laevis Blochwitz, in Ann.
Mycol. 32(1/2): 84. 1934 175

A. ustilagu Beck, in Itin. Prin. S.
Coburgi. 2: 148 (Wien). 1888.
Saccardo, in Sylloge Fungorum.

10:526 (Patavii). 1892 223

Syn. A. phoenicis in the A. niger
group 222

A. Vancampenhouti Mattlet, in
Ann. Soc. Beige Med. Trop. 4:
167-176. 1924. Name only
ibid. 6: 31. 1926. In the A.
versicolor group 193

S. varia Bainier, in Bui. Soc. Bot.
France 27: 30. 1880. Probably
some nonascosporic A. nidulans,
or A. unguis.

A. variabilis Gasperini, in Atti Soc.
Toscana Nat. Sci. Pisa, Mem.
8, fasc. 2, p. 326. 1887. From
the description, this was proba-
bly some strain of the A. flavus
group 266

A. varians Ceni, in Ri vista speri-
mentale di Freniatria, p. 31.
1905. Certainly identified by
Tiraboschi in Annali di Bot. 7:
9-10, 1908, as A. versicolor.

A. varians Wehmer, in Bot. Centralb.
80: 460-1. 1899. Also in Weh-
mer Monogr. pp. 77-79, taf. I.
1899-1901. If Wehmer's de-
scription is correct, his species is
not known in culture now. The
authors have believed that A.
varians is identical with A.
itaconicus 142

A. variecolor (Berk, and Br.) Thorn
and Raper, in Mycologia 31: 663-

667, fig. 4D and fig. 5. 1939 163

Syn. Emericella variecolor Berk,
and Br., in Introd. Crypt. Bot.
p. 340-341, fig. 76. 1857. See
Patouillard in Bull. Soc. Myc.
Fr. 7: 43-49, pi. 4, fig. 6-12.

1891 163

Syn. Inzengaea erythrospora Borzi,
Jahrb. Wiss. Bot. (Pringsheim)

16: 450-463, pi. 19, 20 (1884).

1885 163

Syn. Emericella medias Chowdhury
& Mathur. Ann. Myc. 36: 61-63.

1938 163

Syn. .4. stellatus Curzi, Rend.
Acad. Naz. Lincei 19, p. 424-428,
fig. 1. 1934 163

A. variegatus Mosseray, in La Cellule
XLIII: 238-239, pi. 4, fig. 72-75.
1934. Member of the A. niger
group; in the Biourge Collection.
1939 233

A. velutinus Mosseray, in La Cellule
XLIII: 252-253, pi. 3, fig. 38-41,
1934. Member of the A. niger
group; in the Biourge Collection.
1939 234

S. veneta Massalongo, in Bui. Soc.
Bot. Ital. No. 7-8, p. 159. 1900.
Not cultivated and not identifi-
able by description, although
Werkenthin (Phytopathology 6:
247-249. 1916) used A. venetus
for strains now known to be A.
terreus.

E. verruculosum Vuillemin, in Bui.
Soc. Myc. France 34: 83. 1918.
See A. echinulatus in A. glaucus
group 131

A. versicolor (Vuillemin) Tiraboschi,

in Ann. Bot. (Rome) 7: 9. 1908. 190
Syn. <S. versicolor Vuillemin, cited
by Mirsky, in These de Med.
Nancy No. 27, p. 16. 1903. A.

versicolor series 190

mut. coerulea Blochwitz, in Ann.
Mycol. 27(3/4): 201. 1929. Rep-
resents a pure blue strain arising
from a green strain. Probably
A. sydowi.
var. fulvus Nakazawa, Takeda and
Suematu, culture available from
the Centraalbureau, 1939.
var. glauca Blochwitz, in Ann.
Mycol. 32: 86. 1934. A strain
greener than typical for the
species, from human skin accom-
panying a Trichophyton. Simi-
lar green strains have been
observed by us 192

CHECK LIST OF SPECIES AND GENERA

359

A. violaceo, cited by Biourge in his
MS. p. 16, but no data offered.
.4. violaceo-fuscens (Was this A.
violaceo-fuscus?) mut. grisea
Blochwitz, in Ann. Mycol. 32:
87. 1934. The grounds of sepa-
ration are vague.
A. violaceo-fuscus Gasperini, in Atti
Soc. Toscana Sci. Nat. Pisa,
Mem. 8, fasc. 2, p. 326. 1887.

In .4. niger group 231

A. virens Link, in Obs. Ord. PI. Nat.
p. 16, 1809; also Sp. PL, Ed. 4, 6:
pt. 1, p. 67. 1824. This has
never been surely identified.
A. virens (Link?) rLichelbaum, in
Verh. Naturw. Ver. Hamburg 3
Folge. XIV. p. 34. 1906. This
might have been close to A.
fischeri.
Syn. E. virens (Eichelb.) Sacc.
Saccardo cites description of
preceding under this name in
Sylloge Fung. 22: 1255. 1913.
A. riridans nomen nudum attached
by Biourge to a culture distrib-
uted by him. One of the A.

rest rictus series 1-10

.4. riridis apparently undescribed,
appears only on a specimen in
the Curtis Collection now in the
Cryptogamic Herbarium of
Harvard University; the label is
"A. riridis Schw. in herb., A.
virens Lk. in Syn.: U. S., Herb.
Schw." No Aspergillus could
be found in the specimen.
,4. rirido-griseus Costantin and
Lucet, in Ann. Sci. Nat. Ser. IX,
2: 140. 1905. One of the A.

fumigatus group 151

S. ritellina Ridley, in Jour. Bot. 34:
152, Plate 357, figs. 14-16. 1896.
No one has since reported Kid-
ley's organism with certainty
but a culture belonging in the A .
ochraceus group appears under

the name in C. B. S. list. Bloch-
witz regarded it as a synonym for
A. flavus 276

.4. oulpinus nomen nudum was dis-
tributed by Biourge. It be-
longed to A. tamarii Kita 256

S. welwitschiae (Bresadola) Hen-
nings. Cited by Wehmer in
Centralb. f. Bakt. etc., 2 Abt . 18:
394-395. 1907. Examination of
material from Hennings by Weh-
mer furnished no basis for sepa-
rating the form from A. niger.
Examination of similar specimen
in the Farlow Herbarium shows
primary sterigmata up to 50 by
12m which would place it near A.
phoenicis.
Syn. Ustilago welwitschiae Bresa-
dola.

A. wehmeri Constantin and Lucet,
in Ann. Sci. Nat. Bot. Ser. IX,
2: 162. 1905. The name is pro-
posed by Costantin and Lucet
for the organism which Brefeld
and Wehmer described as A . fla-
vus Link and which is so used in
this paper. The uncertainties in
the identification of Link's
species do not seem important
enough to justify the change of
name. A. wehmeri is to be re-
garded as a synonym of A.

flavus q.v 266

A. wentii Wehmer, in Centralbl.
Bakt. 2 Abt., 2: 150. 1895. See
also Wehmer, Die Pilzgattung
Aspergillus, etc., in Mem. Soc.
Phys. d'Hist. Nat. Geneva 33:

part 2, p. 119. 1899-1901 246

var. minimus Nakazawa, Takeda,
Okada, and Simo, culture avail-
able from the Centraalbureau in

1939 249

A. westendorpii Sacc. and March, in
Rev. Mycol. 7: 149. 1885. In
A. clavatus group 98

Chapter XXV

ACCEPTED SPECIES, VARIETIES, AND

MUTATIONS

A. alliaceus Thom and Church

A. amstelodami (Mang.) Thom and
Raper

A. atropurpureus Zimmerman

A. avenaceus G. Smith

A. awamori Nakazawa

A. butyracea Bainier

A. caespitosus Raper and Thom. . .

A . candidus Link

A. carbonarius (Bain.) Thom

A. carneus (v. Tiegh.) Bloch., emend.

.4. carnoyi (Biourge) Thom and
Raper

A. chevalieri (Mang.) Thom and
Church

A. chevalieri (Mang.) Thom and Ch.
var. intermedins Thom and Ra-
per

A. citrisporus von Hohnel

A . clavatus Desm

A. conicus Blochwitz

A. delacroixii (Sacc.) Thom and
Church

A. echinulatus (Delacr.) Thom and
Church

A. effusus Tiraboschi

A. elegans Gasperini

A. fischeri Wehmer

A. flavipes (Bain, and Sart.) Thom
and Church

A . flavus Link

A. foetidus n. sp

A. fonsecaeus n. sp

A. fumaricus Wehmer

A. fumigatus Fresenius

A. fumigatus (Fres.) mut. helvola
Yuill

A. giganteus Wehmer

A . gracilis Bainier

A. granulosus Raper and Thom

A. humicola Chaudhuri and Sachar.

A. itaconicus Kinoshita

A. jarius Raper and Thom

244

122
226
246
220
282
166
207
229
201

282

131
267
281
151

179
263
219
227
227
148

150
95
138
175
193
142
187

A
A
A
A
A
A
A
A

A
A

134 A
A

118

A

121 A

251 A
92

140 A

A
A
A
A
A
A
A
A
A
A
A
A
A

A
A
A

A
A

janus var. brevis Raper and Thom 190

japonicus Saito 231

luchuensis Inui 230

lutescens (Bain.), Thom & Church 251

mangini n. comb 127

medius Meissner 133

melleus Yukawa 279

micro-virido-citrinus Cost, and

Lucet 263

miyakoensis Nak., Simo & Wat.. 220
montevidensis Talice and Mac-
Kinnon 125

nidulans (Eidam) Wint 156

nidulans (Eidam) Wint. mut. alba

Yuill 159

nidulans (Eidam) Wint. var.

latus Thom and Raper 159

niger van Tieghem 216

niger v. Tiegh. mut. cinnamomeus

(Schiem.) n. comb 223

niger v. Tiegh. mut. schiemanni

(Schiem.) n. comb 224

niveo-glaucus Thom and Raper. . 135

niveus Bloch., emend 202

ochraceus Wilhelm 279

oryzae (Ahlburg) Cohn 261

ostia7ius Wehmer 283

panamensis Raper and Thom. . . . 242

parasiticus Speare 266

. penicilloides Speggazzini 142

penicillopsis (Hennings) Racib.. 282

phoenicis (Cda.) Thom 222

proliferans G. Smith 117

. pseudoglaucus Bloch 110

, pulchellus (Speg.) Thom and

Church 228

. pulverulentus (McAlpine) Thom 223
. quadrilineatus Thom and Raper 160
. quercinus (Bain.) Thom and

Church 276

. repens (Cda.) DeBary 103

. restrictus G. Smith 141

360

ACCEPTED SPECIES, VARIETIES, AND MUTATIONS

361

A. ruber (Brem.) 114

A. rugulosus Thorn and Raper 160

A . sclerotiorum Huber 278

A. sparsus Raper and Thorn 283

A. sulphureus (Fres.) Thorn and

Church 275

A. sydowi (Bain, and Sart.) Thorn

and Church 184

A. tamarii Kita 254

A . terreus Thorn 195

A. terreus Thorn var. aureus n. var.. 198
A. terreus Thom var. boedijni n. var. 197
A. terreus Thom var. floccosus Shih 198

A . terricola Marchal 253

A. terricola var. americana Marchal 253
A. umbrosus Bainier and Sartory. . . 129
A. unguis (Emile-Weil and Gaudin)

Thom and Raper 169

A. ustus (Bainier) Thom and Church 171
A. ustus (Bain.) Thorn and Ch. var.

laevis Blochwitz, n. comb 175

A. variecolor (Berk, and Br.) Thom

and Raper 163

A. versicolor (Vuill.) Tiraboschi .... 190

A . violaceo-fuscus Gasperini 231

A , wentii Wehmer 246

INDEX

Abbott, E. V., 174, 175, 319

Accepted species, 360
Acid production

Aconitic, 299

Amino, 299

Aspergillic, 272

Citric, 237, 249, 290-293

Fumaric, 237, 293-294

Gallic, 237, 294

General, 294-295

Glaucic,299

Gluconic, 238, 295-297

d-Gluconic, 299

Glucuronic, 299

Glycolic,299

Glyoxylic,299

Itaconic, 143, 204-205, 297

Kojic, 249, 258, 270, 297-298

Malic, 299

Melleic,299

Oxalic, 238, 298-299
Aconitic acid, 299
Actinomycetes, 42, 31
Agar slant cultures, 51
Albino forms, 206, 212
Alcohol, 316

Alcoholic fermentations, 270
Alexander, D. F., ix
Alkaline reaction by A. clavatus, 95
Allergy, 154, 271
Alliospora, 9
Alphabetical check list of species and

genera, 330
Amann, J., 47, 319
Amino acid formation, 299
Anastomoses, 65
Anslow and Raistrick, 154, 319
Antibiotics, 98

from A. clavatus, 98

from A. giganteus, 99

from A.flavipes group, 182

from A. flavus-oryzae group, 272
from A.fumigatus, 154
Arlington Farm, 227
Ascogone, 27
Ascomycetes, 6
Ascophora nigrans, 9
Ascospore, 28

Color in A. nidulans group, 28, 156
Germination, 28, 29
Markings, 108, 130, 152, 162
in A. fischeri, 162
in A. glaucus group, 108, 130
in A. nidulans group, 162
Aspergillaceae, 6
Aspergilleae, 6
Aspergillic acid, 272
Aspergilline, 22
Aspergillopsis Sopp, 9
Aspergillopsis Spegazzini, 8
Aspergillosis, in birds, 148, 154
Aspergillus Micheli, 6, 7
Aspergillus, Generic diagnosis, 7
Aspergillus species, *see also Check list

of species, p. 331
A. alliaceus Thorn and Church, 244*, 246,

245, 249
A. amstelodami (Mangin) Thorn and

Church, 122*, 124, 106, 108, 123
A. archiflaripes Bloch., 181
A. atropurpureus Zimm., 226*
A. avenaceus Geo. Smith, 246*, 243, 249
A. awamori Nakazawa, 220*
A. butyracea (Bainier) n. comb., 282*
A. caespitosus Raper and Thorn, 166*-

168, 167
A. candidus Link, 207*-212, 208, 209
A. capitatus ochroleucus Micheli, 3
A. capitulo pulla Micheli, 3, 214
A. carbonarius (Bainier) Thorn, 229*-

230, 217, 218, 236

* Names are cited in this index for recognized or historically important species
only. For a complete list of published names for the Aspergilli, see the Check
List of Species, Chapter XXIV.

363

364

INDEX

A. carneus (van Tiegh.) Bloch., 201*-

202, 199, 200
A. carnoyi (Biourge) Thorn and Raper,

134*-135
A. chevalieri (Mangin) Thorn and

Church, 118M20, 104, 106, 108, 119
A. chevalieri (Mangin) var. intermedins

Thorn and Raper, 121*, 119
A. citrisporus von Hohnel, 251*, 51
A. clavatus Desm., 92*-95, 93, 94
A. conicus Blochwitz, 140*
A. delacroixii (Sacc.) Thorn and Church,

282*
A. echinulatus (Delacr.) Thorn and

Church, 131*-132, 128, 130, 106, 109
A. effusus Tiraboschi, 267*-269, 268, 69
A. elegans Gasperini, 281*
A.fischeri Wehmer, 151M53, 149, 152
A. flavipes (Bainier and Sartory) Thorn

and Church, 179 *-l 81, 180
A.flavus Link, 263*-266, 264, 265, 69
A.foetidus n. sp., 219*, 217, 40
A.fonsecaeus n. sp., 227*-228, 76, 221
A.fumaricus Wehmer, 226*
A.fumigatus Fresenius, 68, 148M51, 149
A.fumigatus (Fres.) var. helvola, 65, 72,

150
A. giganteus Wehmer, 95*-97, 96, 239
A. glaucus Link, 3, 4, 101
A. gracilis Bainier, 138*-139
A. granulosus Raper and Thorn, 175*-

178, 176, 177
E. herbariorum, 4

A. humicola Chaudhuri and Sachar, 193*
A. itaconicus Kinoshita, 142*-143, 109
A. janus Raper and Thorn, 45, 92, 187*-

190, 188, 46
A. janus var. brevis Raper and Thorn,

190*
A. japonicus Saito, 230*
A. luchuensis Inui, 230*
A. lutescens (Bain.) Thom and Church,

251*-253,252
A. mangini (Mangin) n. comb., 127*-129,

128
A. medius Meiss., 133*-134, 46, 128
A. melleus Yukawa, 279*
A. micro-virido-citrinus Cost, and Lucet,

263*
A. miyakoensis Nak., Simo, and Wat.,

220-221

A. montevidensis Talice and MacKinnon,

125*, 123
A. nidulans (Eidam) Wint. 156*-159,

157, 158, 161, 162
A. nidulans mut. alba Yuill, 65, 72, 158,

159*
A. nidulans var. latus Thom and Raper,

159*
A. niger van Tieghem, 216*, 217, 218, 221,

222, 18, 72, 239
A. niger mut. cinnamomeus (Schiem.) n.

comb., 73, 74, 206, 223*-224, 244, 242,

236
A. niger mut. schiemanni (Schiem.) n.

comb., 73, 74, 206, 224*-225, 217, 244,

242, 236
A. niveo-glaucus Thom and Raper, 135*-

137, 19, 128, 136
A. niveus Bloch., emend. 202*-204, 199,

200
A. ochraceus Wilhelm, 279*-281, 280
A. oryzae (Ahlburg) Cohn, 261*-263, 262,

264, 69
A. ostianus Wehmer, 283*
A. panamensis Raper and Thom, 242*-

244, 243, 249
A. parasiticus Speare, 266*-267, 268, 69
A. penicilloides Speg., 142*
A. penicillopsis (Hennings) Racib.,282*-

283
A. phoenicis (Cda.) Thom, 222*, 223,

218, 239
A. proliferans G. Smith, 117*
A. pseudo -glaucus Bloch., 110*— 111 , 105
A. pulchellus (Speg.) Thom and Church,

228*
A. pulverulentus (McAlpine) Thom, 223*
A. quadrilineatus Thom and Raper, 160*

161,162
A. quercinus (Bainier) Thom and

Church, 276*-278, 277
A. repens (Cda.) De Bary, 103M09, 104,

105, 106, 108, 109
A. restrictus G. Smith, 141*, 136, 139, 109
A. ruber (Bremer) n. comb, 114*-117, 104,

106, 108, 113, 115
A. rugulosus Thom and Raper, 160M63,

158, 161, 162
A. sclerotiorum Huber, 278*-279, 277
A. sparsus Raper and Thom, 283*-
285, 284

INDEX

365

A. sulphureus (Fres.) Thorn and Church,

275*-276
A. sydowi (Bain, and Sart.) Thorn and

Church, 184M86, 185
A. tamarii Kita, 254*-257
A. terreus Thorn, 66, 195M97, 196, 199,

200
A. terreus var. aureus n. var., 66, 67,

198*, 199
A. terreus var. boedijni (Bloch) n. comb.,

66, 67, 197*
A. terreus var. floccosus Shih, 66, 67, 198*
A. terricola Marchal, 253*
A. terricola var. americana Marchal,

253*-254,252
A. umbrosus Bain, and Sart., 129*— 131 ,

128, 130
A. unguis (Emile-Weil and Gaudin)

Thorn and Raper, 169M70
A. ustus (Bainier) Thorn and Church,

171M75, 172, 173, 176
A. ustus var. laevis Bloch., 175*, 173
A. rariecolor (Berk, and Br.) Thorn and

Raper, 163M66, 158, 161, 162, 164
A. versicolor (Vuill.) Tiraboschi, 190*-

193, 185, 191
A. violaceo-fuscus Gasperini, 231*, 217,

218
A. wentii Wehmer, 246*-247, 248, 249
Assumptions, Basic, 10

B

Baarn, Holland, — See Centraalbureau
Bacillus anlhracis, 154
Bacteria in Aspergillus cultures, 59
Bactericidal agents — See Antibiotics
Bainier, G., vii, 4, 174, 227, 229, 236, 278
Bainier and Sartory, 107, 319, 114, 129,

133
Barham and Smits, 270, 298
Barnes, B., 74, 144, 145
Barthel, C, 56

Bary, A. De, vii, 4, 101, 26, 28, 7, 27, 103
Basidia, 23

Berkeley, M. J., 163, 165
Bernhauer, K. I., 237, 293, 295, 299, 320,

291,238,306
Bernton,H.S.,154,320
van Beyma, F. H., viii

Bibliographies, types included, viii
Check list of species with bibliographic

references, 331-359
General, viii, 319-330
Topical, viii, 289-318

Bichloride of Mercury, 61

Binocular, wide-field, 40, 45, 51

Biourge, Ph., ix, 4, 135, 139, 214, 232, 235

Bisby,G.R.,ix

"Black Aspergilli" — see A. niger group

Blakeslee, A. F., 41, 320, 229, 230

"Blase", 22

Blochwitz, Adalbert, ix, 4, 64, 66, 137,
175, 197, 202, 203, 206, 229, 231, 236,
256, 267, 279

Bonner, J. T., 242

Borzi, A.,165, 163,321

Brant, Nancy, ix

Brefeld,0.,4

Bulb disease, 246, 249

Buller, A.H.R.,65,321

Bush and Goth, 272, 300

Calam, Oxford and Raistrick, 204, 321
Candidus group, 206-213

Group relationships, 212

Occurrence, 213

Other white Aspergilli, 206, 207, 212

Outstanding characters, 206

Sclerotia, 212

Variation in head size, 209
Capitulum, 3
Carbon dixoide ice, 53
Cardozo, D.M.,228
Cathode rays, 75
Cellulose, decomposition, 150
Centraalbureau voor Schimmelcultures,

viii, 231, 247, 261, 166
Chaetomiwn, 239
Challenger etal., 270, 197
Characters, Diagnostic, 82
Chemistry of Mold Tissue, 301-302
Chitin, 317
Chlamydospores, 28
Chowdhuri and Mathur, 163, 321
Christensen, L. M., 271, 304
Chromotaxia, Saccardo, 242, 273
Church, M. B., ix
Ciferri,R., 166,321

366

INDEX

Citric acid, 237, 249, 290-293

from A . niger, 237. See also 290-293

from A . wentii, 249
Citrinin, 205

Cladosarum olivaceum Yuills, 9, 65, 73,
240

Barnes "Creamy", 74, 145

Conidium formation, 73

Morphology of, 73

Nuclear behavior, 73, 74
Clarke, F. E., 198, 199
Classification, 6
Clavacin, 98
Clavatin, 99
Clavatus group, 92-99

Antibiotics, 98-99

Occurrence, 98

Outstanding characters, 92

Synonyms, 98
Coffee fermentation, 286
Coghill, R. D., ix, 75, 326
Colony characters, 11-16
Colony types, 11-13,12
Color, 13

in conidial walls, 14

in conidiophores, 14, 22, 148, 153, 171,
179, 273

as group characters, 13

in the mycelium, 13

influence of pH, 14

in the substratum, 15
Color photographs of Aspergilli, Plates
I-VII

of A . alliaceus Thorn and Church, Plate
VI, D

of A. amstelodami (Mang.) Thorn and
Church, Plate III, E

of A. avenaceus Smith, Plate VI, E

of A. candidus Link, Plate VI, A

of A. carneus (v. Tiegh.) Blochwitz,
Plate V, F

of A. clavatus Desm., Plate III, A

of A, flavipes (Bain, and Sart.) Thorn
and Church, Plate IV, F

of A. flavus Link, Plate VII, D

of A. fumigatus Fres., PlatelV, B

of A. giganteus Wehmer, Plate III, B

of A. janus Raper and Thorn, Plate I,
E and F; Plate V, B

of A. nidulans (Eidam) Wint., Plate
I,A-D;PlateIV, C

Color photographs of Aspergilli — Cont'd.
of A. niger group, N.R.R.L. 67, Plate

VI, B
of A. niger group, tan-spored mutant

Plate VI, C
of A. niveo-glaucus Thorn and Raper,

Plate III, F
of A. ochraceus Wilhelm, Plate VII, F
of A. oryzae (Ahlb.)Cohn, Plate VII, C
of A. quercinus (Bain.) Thorn and

Church, Plate VII, E
of A. repens (Cda.) De Bary, Plate III,

C
of A. restrictus Smith, Plate IV, A
of A. ruber (Bremer), Plate III, D
of A. sydoivi (Bain, and Sart.) Thorn

and Church, Plate V, A
of A. tamarii Kita, Plate VII, B
of A. terricola var. americana Marchal,

Plate VII, A
of .4. terreus Thorn (unirradiated),

Plate II, A; Plate V, E
of A. terreus (U-V mutants), Plate II,

B-F
of A. ustus (Bain.) Thom and Church,

Plate IV, E
of A. variecolor (Berk, and Br.) Thom

and Raper, Plate IV, D
of A. versicolor (Vuill.) Tiraboschi,

Plate V, C and D
of A. wentii Wehmer, Plate VI, F
Conidia, 18, 19, 24
Coloration, 13
Endogenous, 25
Formation, 23, 24
Germination, 29
Nuclear behavior, 23
Conidiophore, or stalk, 17, 18, 19
Color, 22
Surface, 19
Connective, 25
Contaminants, 57-62
Bacteria, 59

Elimination of, 41-42, 58, 59
Mold disease, 59, GO
Other molds, 58
Recognition of, 58
Cook and Lacey, 272, 301
Corda, A.C. I., 3, 223
Coremia, 13
Cramer, C, 4, 239, 321

INDEX

367

Culture Collections, 50

American Type Culture Collection,
Washington, 50

Centraalbureau voor schimmelcul-
tures, Baarn, Holland, 50

National Type Culture Collection,
London, 50

Northern Regional Research Labora-
tory, Peoria, 111., 50

Thorn Collection, 50
Culture media, See Media
Culture, purification of, 41, 42
Cultures, types of, 38

Dilution cultures, 40

Hanging drop, 45

Single-colony inoculation, 39, 40

Single spore cultures, 42

Spot inoculation, 39

Streak cultures, 41

Three-point inoculation, 39, 40
Currie, J.N. ,237, 328
Curzi, M., 163, 166, 321
Czapek's solution agar, 32

D

Dale, Elizabeth, 27, 101 , 321
Dangeard, P. A., 27, 322, 28, 30
De Bary, A., See Bary, A. De
Descriptive sheet, 82
Descriptive terms, 11
Desiccation of molds, vacuum, 53
Dewar flask, 53, 54
Diastatic enzymes, 259, 257
"Digestin", 213
Dilution cultures, 40
Dimargaris, 9

Diploslephanus Langeron, 8
Disjunctor, 25
Dodge, B. O., Ill
Dodge, C. W., 5, 154, 322
Doelger and Prescott, 237
Dox,A.W.,239,322,32
Dried cultures — See Lyophil preserva-
tion of molds
Dried specimens, 62
Drierite, 53
Dual character of A. janus, 188, 189

E

Earle, F. R., 178
Ecads, 64

Eidam,E.,155,156,322

"Eidamsche blasen", 28

Elser, W. J., et al., 53, 322

Emericella variecolor Berk, and Br., 9,

163
Emmons, C. W.,198
Endogenous conidia, 25
hmgler and Prantl, 6
Enzymes, 238, 249, 213, 257, 270-271, 194,

302-306
Enzyme production

by A . flavus-aryzae group, 270-271 , 302-
304

by A. niger group, 238, 304-305

by A . okazakii, 213

by A . tamarii group, 257

by A . wentii group, 249
Ergosterol, 194,317
Escherichia coli, 154
Euaspergillus Ludwig, 8
Eurotiaceae, 6
Eurotiales, 6
Eurotium Link, 7, 101
Eurotium herbariorum, 101, 125, 127

Fat production, 194, 238, 306-307
Ferdinandsen and Winge, 17, 228, 322
Fermentation Division, N.R.R.L., ix,

205
Fernbach, A., 238, 304
Fish, fermented, 286
Fisher, Ed., 165
Flavicidin, 272
Flavicin, 272
Flavipes group, 179-182

Antibiosis, 182

Color reactions, 180

Occurrence, 182

Outstanding characters, 179
Flavus-oryzae group, 259-272, 69, 68

Antibiosis, 271-272

Enzyme production, 270-271

Kojic acid, 270

"Moldy bran", 271

Occurrence, 269

Outstanding characters, 259

Pathogenesis, 271

Variation in, 68-69, 260
da Fonseca, Olympio, 227, 230, 322

368

INDEX

Foot-cell, 17, 19

of A. effusus, 264
Frazer and Chambers, 27, 322
Fresenius, G., vii, 4, 275, 148
Fumaric acid, 237, 293-294
Fumigacin, 154
Fumigatin, 154
Fumigatus group, 148-154

Allergy, 154

Antibiosis, 154

Economic importance, 153

Occurrence, 153

Outstanding characters, 148

Pathogenicity, 154

Thermophilic habit, 153
Fungi Imperfecti, 6

G

Gallic acid, 238, 294
Galloway, L.D., 75, 322
Gene mutation, 63
General bibliography, 319-330
Generic diagnosis, 6
Gilman and Abbott, 202, 322
Glaucic acid, 299
Glaucus group, 100-147

Economic importance, 146

Group relationships, 102

Historical considerations, 101

Laboratory cultivation, 101

Occurrence, 146

Outstanding characters, 100

Pathogenicity, 146

Pigment formation, 15

Temperature relations of, 45, 133, 134

Variation in, 143-145
Glister, G. A., 272, 300
Gluconic acid, 238, 295-297
d-Gluconic acid, 299
Glucuronic acid, 299
Glycolic acid, 299
Glyoxylic acid, 299
Gould, B.S., 258, 298
Gould and Raistrick, 15, 323
Graphic key to groups, 87
Greene, H. C, 65, 323
Greene and Fred, 56, 323
Group keys

A. candidus group, 207

A. clavatus group, 92

A.flavipes group, 179

A . flavus-oryzae group, 259-260

A. fumigatus group, 148

A. glaucus group, 102

A. nidulans group, 155-156

A. niger group, 215-216

A. niger group (Mosseray's), 232-235

A. ochraceus group, 274-275

A. tamarii group, 250-251

A . terreus group, 195

A. ustus group, 171

A. versicolor group, 183

A. wentii group, 241
Growth substances, 316
Guegen, F.,4, 323
Gum, 317

H

Haines, R. W., ix, Plates I-VII

Haller, D. A.,3

Hanging drop culture, 45

Hansen, H. N., 65, 323

Hansen and Smith, 65, 323

Hanzawa, J., ix, 225

Hao,L. C.,271,303,304

Hay-infusion agar, 35

Head, 16, 18-21

Henrard, P., 27, 323

Herbarium specimens, 62

Herrick, H. T., ix, 237, 238, 227

Heterothallism, 27

High sugar Czapek's agar, 101

Historical introduction, 3

Hollaender, A.,75, 326

Homothallism, 27

Hooper, et al., 99, 323

Huber,G.A.,278,286,323

Hulle cells, 28

in A. carneus, 176, 202

in A . flavipes group, 179

in A. nidulans group, 157, 167

in A. ustus group, 176

in A. versicolor group, 188, 192

Hydrogen-ion concentration, 14, 34, 35,
95

Hydroxylamine, 317

Hyphomycetes, 6

I
Identification of Aspergilli, 81-91
Incubation of stock cultures, 51

INDEX

369

Incubators, 48

Induced variation, 74

Industrial Farm Products Research

Division, 227
Inoculating needles and loops, 47
Intermediate species, 86
Interpretation of Descriptions, 10
Inui, T., 230
Invertase deficiency in A. panamensis,

244
Inzengea erythrospora Borzi, 9
Isolation of single spores, 43, 44
Itaconic acid, 204-205, 143, 297

Jardin Botanique de l'Etat, 235
Jones, Rake, and Hamre, 272, 301

K

Karow,E.O.,249,293,323
"Katsuobushi", 286
Keitt, G. W., 43, 323
Keys, 86-91

based on color, 88-89

based on morphology, 90-91

Graphic, 87

to groups, 86-91

to species — see group keys
Kinoshita, K., 142, 143, 323, 324
Kita,G.,ix,255,260,324
Kluyver and Perquin, 270, 298
Koji,38,249,257,260
Kojic acid, 249, 258, 270, 297-298

Lacto-phenol, 48
Lambert, E. B., 43, 324
Langeron,M.,204, 324
La Rue, CD., 43, 324
Ledingham, G. A., ix, 198
Link, H. F.,3, 101,210
Linossier, G., 206, 324, 15, 236
Lockwood, et al., 76, 324, 205, 297
Loops, transfer, 47
Ludwig, F.,8,324
Lutz,L.,226

Lyophil preservation of bacteria, 53
Lyophil preservation of molds, 53
Advantages of, 56

Apparatus, 54
Methods, 53, 55
Recultivation, 55
Viability, 53

M

Ma, Roberta, ix

Macroaspergilli, 16

Macy, H., ix

Maintenance of cultures, 51-57

Malic acid, 299

Malt extract agar, 35

Mangin, M. L., 118, 122, 127, 143

Mannitol, 317

Manual, use of, 81-91

Marchal,E.,253,324

Martin, G.W., 6

Mary Clare, Sister, 285

May, O. E., ix, 227, 237, 238, 270

McCoy, Prof. Elizabeth, 56

McKee and MacPhillamy, 272, 301

McKee, Rake, and Houck, 272, 301

Mealy bugs, 267, 260

Media, culture, 31

Czapek solution agar, 32

Hay infusion agar, 35

High-sugar Czapek agar, 101

Influence of, 36

Malt extract agar, 35

Mosseray's Raulin's solution, 34

Neutral Raulin's solution, 33

Sporulation, 37, 38

Steep-liquor Czapek's agar, 35

Steinberg's solution, 34
Melleic acid, 299
Metarrhizium, 239
Methyl cellosolve, 53
Micheli, P. A., vii, 3, 100
Microaspergilli, 16

Microbiological Congress, Third Inter-
national, 5
Microscopes, 48
Mildew, 146, 239
Mites, 60

Damage caused by, 61

Elimination of, 61-62

Poison, 61
Mixed cultures, 57
Mold Disease of A . niger, 59, 60
Mold Tissue, Composition of, 301-302

370

INDEX

"Moldy bran", 271

Molliard,M.,238,295

Monilia, 3

Monograph, 5

Monospore Cultures, 42

"Monster", 74, 85

Montagne, J. F., 3

Morphology and Description, 10

Morrow, Marie B., ix

Mosseray, Raoul, ix, 70, 75, 83, 229, 232,

235
Mosseray 's Synopsis of Species, 232-234
Mounting fluid, 47

Moyer, A. J., 37, 227, 237, 238, 258, 270
Moyer and Coghill, 205, 297
M-type and C-type, 65
Mucidinaceae, 6
Mucidineae, 6
Mucoraceae, 42
Mucor herbariorum, 7
Mutant, definition of, 63
Mutation, 63
Mutations, deficiency, 76
Mutations, induced, 74

by cathode rays, 75

by chemicals, 74, 75

by heat stimulation, 74

by ultra-violet radiation, 75, 76, 77

in A. glaucus group, 74

in A. niger, 75

in A. terreus, 75
Mutations, "injury", 75
Mutations, morphological, 76, 77
Mutation, natural, 71

in A . fumigatus, 72

in A. glaucus group, 73

in A. nidulans, 72

in A. niger, 72, 73
Mutations, physiological, 78
Mutation, taxonomic usage, 84
Nakazawa, R., ix, 225
Nakazawa, Simo, and Watanabe, 220

N

Natural groups, 82, 87

Natural Mutation, 71

Natural relationship of groups, 87

Needles, transfer, 47

Neill, J. C, 5, 69, 325, 83, 143, 193

Neutral Raulin's solution, 33

New species, 84

Bases for description, 85

Recognition of, 84
Nidulans group, 155-170

Occurrence, 170

Outstanding characters, 155-156

Pathogenicity, 170
Niger group, 214-240

Citric Acid, 237, 290-293

Coloration, 236

Conidiophore structure, 19, 222

Enzyme production, 238, 304-305

Fat production, 238, 306

Fumaric Acid, 237, 293-294

Gallic Acid, 237, 294

Gluconic Acid, 238, 295-297

Industrial strains, preservation, 240,
50-62

Mildew, 239

Mosseray 's Synopsis of Species, ^32-
234

Occurrence, 236

Outstanding characters, 214

Oxalic acid, 238, 298-299

Pathogenesis, 239-240, 307-310

Physiology, 239, 310-312

Soil analysis, 238, 313-314

Strain "67", 227

Variation in head structure, 218

Variation in spore size, 221
Niklas, H., 238, 313

Northern Regional Research Laboratory
vii,50

O

Ochraceus group, 273-286

Fermentations, 286

Occurrence, 285-286

Outstanding characters, 273
Ochracin, 318
Odor

Actinomyces-like in A. foetidus, 219

foetid in A. clavatus, 95

foetid in A. flavipes, 180
Okazaki,K., 213, 211,325
Oryzae group — See Flavus-oryzae group
Oshima, K., ix, 260, 303, 261, 270
Ota, M., 271,308
Overgrowths, 58
Oxalic acid, 238, 298-299
Oxford and Raistrick, 154, 325

INDEX

371

Parasiticin, 272
Parasitism of Aspergilli, 59
Partansky and McPherson, 239, 325
Pathogenicity, 307-310

in A . flavus-oryzae group, 271

in A. fumigatus group, 154

in .4 . glaucus group, 146-147

in A. nidulans group, 169-170

in .4. niger group, 239-240

in A. terreus group, 204

in A. versicolor group, 193-194
Patouillard and Delacroix, 222, 223
Patulin,98
Penicillin, 99, 182
Penicillin-like substances

in A. flavipes, 182

in A.flavus, 272

in A. giganteus, 66
Penicillium chrysogenum Thorn, 57
Penicillium notatum Westling, 272
Penicillium patulum West., 99
Penicillium restrictum Abbott, 67, 186
Penicillium rugulosum Thorn, 59
Penicillium spinulosum Thorn, 154, 300
Perithecia, 26

in A . fischeri, 149, 151-153

in A. glaucus group, 27, 100-137, 106,
115

in A. nidulans group, 155-166, 161, 164
Persoon, C. H., 3

Pfizer (Chas.) and Co. Inc., ix, Pis. I-VII
Phialids, 23
Philpot, F. J.,99,301
Photographic equipment, 49
Physiology, 239, 310-312
Pigments, 15, 143, 236, 312-313
Plectascineae, 6
Plug poison, 61
Poison, cotton plugs, 61
Polysaccharides, 318
Pontillon, C, 238, 306
Preservation of Cultures. 51

in agar slants, 51-52

in lyophil form, 53-56, 52

in soil, 56-57, 52

on vegetable substrata, 57

Industrial strains, 240, 50-62
Progressive variation, 68

Proteolytic enzyme, 213, 257, 271
Pulli, 214

Q

Quilico and Di Capua, 15, 313

R

Raistrick, Harold, ix, 111, 117, 143, 231,

238, 283
Raistrick, Robinson and Todd, 15, 313
Rami, 23
Raper, Coghill, and Hollaender, 75, 326,

205, 206, 212, 225
Raper and Thorn, 166, 175, 187, 190, 242,

283
Raulin, J.,4,237
"Raulin neutre-gelose", 70, 33
Raulin's solution, 34
Recultivation of dried cultures, 55
Reduced structures in A. sydowi, 184
Rejected species — See Check list of

species
Resting nuclei, 73
Ridgway, Robert, 13, 326
Ridley, H. N., 13,64,326

S

Saccardo, P. A., 11, 165, 326

Saccharification, 271

Saito,K.,231,297

Salmonella typhi-murinum, 154

Saltant, 64

Sartory , A . , 4

Sartory and Meyer, 212, 327

Sartorya fumigata , 9, 153

Scales, F. M., 253

Schiemann, Elizabeth, 74, 223, 224, 327,

206
Schmidt, C.F., Jr., 238, 306
Schwartz, W., 30, 327
Sclerotia, 30, 212

in A. candidus group, 212, 208

in A. flavus-oryzae group, 259, 265

in A. niger group, 214, 217

in A. ochraceus group, 273, 277

in A . tamarii group, 250

in A . wentii group, 241 , 243

significance of, 212

structure of, 30
Secondary growth, 58

372

INDEX

Sectors, 64

Septa, 17

Shih, Y. K.,ix,198

Short-lived species, 51

Siebenmann, F. , 271 , 307

Simonart, Paul, ix, 232

Single-spore cultures, 42, 44

Smear cultures, 41

Smith, George, ix, 5, 117, 102, 137, 139,

141,246,285
Sodium lauryl sulfonate, 37
Soil analysis, 238, 313-314
Soil Fungi, Summary of, 202
Soil Preservation of Molds, 56

Directions, for, 56

Viability, 56
Sopp,0. J.O.,9
Soy products, including soya sauce, 249,

258, 270
Speare,A.T.,260,266,328
Species

Accepted, 360-361

Check list of, viii, 331-359

Definition of, 83

Diagnosis of, 82

Keys to, see group keys

New, 84

Rejected — See Check list of species

Transitional, 86
Specimens, identification from, 81
Spegazzini,C.,8,229,328
Spinulosin, 154
Spores

Ascospores, 28, 29, 152, 162, 130, 108

Conidia, 23-25, 24, 221
Sporodinia, 3
Sporulation media, 37, 38
Sprays, 62

Stalk — See Conidiophore, 17
Staphylococcus, 53, 98, 99
Staphylococcus aureus, 154, 182
Steep liquor Czapek's agar, 35
Steinberg, R. A., 239, 310, 311, 312
Steinberg's nutrient solution, 34
Steinberg and Thorn, 75, 328, 206, 2^2,

224, 225, 240
Sterigmata, 22

Primary, 19, 23

Secondary, 19,23
Sterigmatocystis Cramer, 8
"Sterile hyphae" in A. unguis, 169

Stock cultures, 51-57
Agar slants, 51
Cultivation, 51
Incubation, 51
Periodic transfer, 51
Storage, 51

Streak cultures, 41

Streptococcus viridans, 154

Substrata, see Media

Swift, MarjorieE., Ill

Synonyms

in A. candidus group, 211

in A. chevalieri series, 120, 121

in A. clavatus group, 98

in A. echinulatus, 133

in A. flavus-oryzae group, 266, 269

in A. luchuensis series, 232

in A. niger series, 225-226

in A. niveo-glaucus , 137

in A. ochraceus series, 281

in A. repens series, 107, 110

in A. restrictus series, 139

in A . ruber series, 117

in A. sulphureus series, 276

in A. sydoivi series, 186

in A. tamarii group, 256-257

in A. terreus group, 197, 200

in A. umbrosus series, 131

in A. versicolor series, 192, 193

in A. wentii group, 247-248

Synonymy, 6

"System und Phylogenie", 5

Takahashi,T.,260,328
Takamine, J., 270, 302
"Tamari", 258
Tamarii group, 250-258

Economic importance, 257-258

Occurrence, 257

Outstanding characters, 250
Tamiya and Morita, 4
Tannic acid, 237, 294
Tannin, 237, 294
Taubenhaus, J. J., ix
Temperature, 45, 46
Temperature, effect of,

on A. giganteus, 45, 97

in A. glaucus group, 45

on A . janus, 45, 187

on A. medius, 45, 46, 134

INDEX

373

Temperatures, optimum, 45
Terminology, descriptive, 11
Terrein, 318
Terreus group, 195-205

Antibiotics, 205

Itaconic acid, 204, 205, 297

Mutations, 75

Occurrence, 204

Outstanding characters, 195

Pathogenesis, 204

Variation in, 66, 67, 197
Tessar lenses, 49
Thaxter, R. T., 50, 189, 251
Thermophilic species, 45
Thorn, C, vii,5, 195,328
Thorn and Church, vii, 4, 5, 102, 137, 206,

235, 255, 260
Thorn and Raper, 74, 328, 102, 168
Thorn and Steinberg, 75, 328, 144, 150,

212,221
van Tieghem, Ph., vii, 4, 237
Timonin, M. I., 203, 205, 329, 213
Topical bibliography, 289-318
Transfer needles and loops, 47
Transfer of stock cultures, 51
Transitional species, 86
Trichoderma, 42
Turfltt,G.E.,246
"Type" culture collections, 50

U

Ultra-violet radiation, 75, 76, 77
Underkofler et al., 271, 303, 285
Ustilago phoenicis Cda., 223
Ustus group, 171-178

Hulle cells, types, 176

Occurrence, 178

Outstanding characters, 171

V

Vacuum desiccation of molds, 53

Vacuum tester, 55

Variant, 64

Variation, natural, 63-78

in A. fischeri, 65

in A . flavus-oryzae group, 68, 69

in A . fumigatus, 68

in A. niger group, 70, 71

in A . sydowi, 67

in A. terreus group, 66, 67

Intra-group, 68

Intra-species, 66, 67

Intra-strain, 65
Variation, references, 314-315
Variety, taxonomic usage, 84
Vegetable substrata, 57
Versicolor group, 183-194

Occurrence, 194

Outstanding characters, 183

Pathogenesis, 193

Strain variation, 192, 186
Vesicle, 18, 19, 22
Vestigial characters, 82
Vibrio cholorae, 154
Vitamin D, 238
Vitamins, 316
Vuillemin, P.,165,329

W

Waksman and Bugie, 272, 301

Waksman et al., 98, 300, 99, 301, 154

Ward, G.E., 237, 270, 329

Webb, P. H.W.,97,329

Wehmer, C, vii, 4, 226, 237, 238, 247, 283

Weisner, B. P., 98, 300

Wells, May, Moyer, Herrick, et al., 227,

237, 270
Wentii group, 241-249

Economic importance, 249

Occurrence, 249

Outstanding characters, 241
Westerdijk, Johanna, viii, 110, 142, 283
Whelden, R. M., 75, 330, 206, 225
White; E. C, 182, 330, 271, 300, 272
White mutants, 206
Wickerham, L. J., 55
Wickerham and Andreasen, 53, 330
Wiggers, Fredricus Henricus, 7, 100
Wilhelm, K. A., vii, 4, 281, 330
Wilkins and Harris, 99, 300
Wire, nichrome, 47

platinum-iridium, 47
Wolf, F. A., 97, 330

Y

Yabuta, T., 249, 297, 330
Yuill, Edward, 65, 150, 159, 206, 212, 221
Yuill, John and Edward, ix, 73, 145, 240
Yukawa, M., 279, 286, 330

Z

Zea Mays, 267
Zonation, 11-13, 12