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Consumption of cyanogenic bamboo by a newly discovered species of bamboo lemur

Authors:

Abstract

Three species of bamboo-eating lemurs were found to be sympatric in the southeastern rain forests of Madagascar. Sympatric species generally differ in habitat utilization or diet, but these three closely related bamboo lemurs lived in the same habitat and all ate bamboo. Behavioral observation revealed that they did select different parts of the bamboo, and chemical analyses confirmed that there was a difference in the secondary compound content present in those selections. The growing tips of Cephalostachyum ef uiguieri selected by the golden bamboo lemur (Hapalemuraureus) contained 15 mg of cyanide per 100 g fresh weight bamboo while the leaves of C. perrieri selected by the gentle bamboo lemur (H. griseus)and the mature culms of C. cf uiguieri selected by the greater bamboolemur (H. simus) did not contain cyanide. Since each individual golden bamboo lemur ate about 500 g of bamboo per day, they daily ingestedabout 12 times the lethal dose of cyanide. The mechanism by which this small primate avoids the acute and chronic symptoms of cyanide poisioning is unknown.
American
Journal of
Primatology
19:119-124 (1989)
BRIEF REPORTS
Consumption
of
Cyanogenic Bamboo by a Newly
Discovered Species
of
Bamboo Lemur
KENNETH E. GLANDER', PATRICIA
C.
WRIGHT", DAVID
S.
SEIGLER:', VOARA
RANDRIANASOLO',
AND
BODOVOLOLONA RANDRIANASOLO'.
'"Department
of
Biological Anthropology and Anatomy, Duke Uniuersity, Durham, North
Carolina; Department
of
Biology, California Institute
of
Technology, Pasadena;
.'Department
of
Plant Biology, Uniuersity of Illinois, Urbana; 'Parc Botanique et Zoologique
cle
Tsinibazaza, Antananariuo, Democratic Republic
of
Madagascar
Three species of bamboo-eating lemurs were found to be sympatric in the
southeastern rain forests of Madagascar. Sympatric species generally dif-
fer in habitat utilization or diet, but these three closely related bamboo
lemurs lived in the same habitat and all ate bamboo. Behavioral observa-
tion revealed that they did select different parts of the bamboo, and chem-
ical analyses confirmed that there was a difference in the secondary
compound content present in those selections. The growing tips of
Ceph-
alostachyum
cf
uiguieri
selected by the golden bamboo lemur
(Hapalemur
aureus)
contained 15 mg of cyanide per 100 g fresh weight bamboo while
the leaves of
C.
perrieri
selected by the gentle bamboo lemur
(H. griseus)
and the mature culms of
C.
cf
uiguieri
selected by the greater bamboo
lemur
(H. simus)
did not contain cyanide. Since each individual golden
bamboo lemur ate about
500
g
of bamboo per day, they daily ingested
about 12 times the lethal dose of cyanide. The mechanism by which this
small primate avoids the acute and chronic symptoms of cyanide poision-
ing is unknown.
Key words:
Hapalemur aureus, Hapalemur griseus, Hapalemur simus,
INTRODUCTION
In 1986, while studying the gentle bamboo lemur
(Hapalemur griseus)
in the
Ranomafana National Park, Madagascar, Wright and colleagues [Wright
et
al.,
1987; Meier et al., 19871 discovered a new species of bamboo lemur
(H. aureus,
or
golden bamboo lemur) and found another species that had not been seen since the
early 1970s
(H. simus,
or greater bamboo lemur). All three of these lemurs live in
the same habitat and all three eat bamboo, in the case of the golden and greater
bamboo lemur, the same species of bamboo [Wright et
al.,
19871. Here we report the
preliminary results of an ongoing behavioral and ecological study of these three
bamboo-eating lemurs.
.
cyanide,
diet,
niche
separation
Received for publication February
6,
1989;
revision accepted August
2,
1989
Address reprint requests to K.E. Glander, Department
of
Biological Anthropology and Anatomy,
Wheeler Building,
3705B
Erwin
Rd.,
Duke University, Durham, NC
27705.
0
1989
Alan
R.
Liss,
Inc.
120
I
Glander et
al.
MATERIALS AND METHODS
Observation
Golden bamboo lemurs were observed for a total of 153 hours during June-
August 1986; June-November 1987; and June-November 1988. Marked individ-
uals were followed for as long as they could be kept in sight. Activities were
recorded, and if the activity was feeding, the part of the plant eaten was noted.
Collection
of
Weights
Capture ofH.
griseus
and
H.
aureus
individuals was accomplished in May 1988
using the Pneu-dart system (Pneu-Dart Inc.,
HC
31,
Williamsport, PA 17701). This
system employs disposable non-barbed darts with a 318-inch needle delivered by a
carbon dioxide powered gun. The darts were loaded with
0.4
cc
(40
mg) of Ketalar
(Ketamine hydrochloride, 100 mglml) (Bristol Laboratories, Syracuse, NY 13201).
Collection
of
Plant Material
Samples of two species of bamboo,
Cephalostachyum
cf
uiguieri
and
Cephalos-
tachyum perrieri,
were collected and tested in May 1987; May, June, and December
1988; and May 1989. The four parts of each plant collected were the leaf, new shoot,
young stem pith, and trunk. Collections
were
made at the exact location where
individuals were feeding or the material dropped was collected in an effort to
obtain plant material as similar as possible to what the animals ate.
The bamboo genus
Cephalostachyum
occurs in Indomalaysia and Madagascar
[Airy Shaw, 19731. Both
Cephalostachyum
species in this study are native to this
area of Madagascar and there are at least 10 species of bamboo in the park area
(Edelman personal communication). This group of grasses is poorly known and the
systematic status of these two species and the genus
Cephalostachyum
is unclear.
Thus the identification of the bamboo in this paper as
C. uiguieri
must be consid-
ered provisional.
Chemical Analyses
The presence or absence of cyanide was determined by the Feigl Anger test
[Feigl
&
Anger, 1966; Tantisewie et al., 19691. All analyses were started and
completed in the shade, at the same temperature and humidity. All tests were done
in the field in May 1987; May, June, and December 1988; and May 1989. The plant
parts tested were chopped into 1-cm-long bits and added to glass test tubes. The
material was macerated in the bottom of vials before a strip of filter paper im-
pregnated with Feigl Anger solution [Feigl
&
Anger, 19661 was added and a cork
placed in the top of the vials. Care was taken
so
that the paper did not touch the
plant material. The white strip changed to blue in the presence of cyanide. The
color of the test strips was noted at intervals of 15, 30, 60, and 240 minutes.
Replicates (a minimum of 3-4 from each plant) of leaves, young stems, and mature
stems of each species were examined.
To determine the amount of cyanide present, the outer fibrous material of the
bamboo was removed from approximately the last foot of growth collected on May
20, 1987. The white tender inner material (115 g) was cut into small pieces as
quickly as possible and added to the hot alcoholic extraction medium (Saint
Claude, Rhum de Luxe, Extra Fin, Special 43", Carte Noire). The mixture was
boiled for 10 minutes, cooled, and decanted. After boiling, the residual plant ma-
terial still contained residual cyanogenic materials as indicated by tests with Feigl
Anger paper [Tantisewie et al., 1969; Feigl
&
Anger, 19661. This material was
reboiled with 200 ml water for an additional 10 minutes. It still gave positive tests
Lemur Niche Separation
I
121
for cyanide, and it is clear that the extraction under field conditions was ineffi-
cient. The percent of cyanide was later determined in the laboratory.
Cyanogenic compounds in the extract were decomposed by addition of emulsin
(Sigma,
St.
Louis, MO) and the resulting cyanide was captured in 0.1 N sodium
hydroxide solution in Warburg flasks. The basic solution was removed and cyanide
content determined by colorimetric analysis [Lambert et al., 19751. Each sample
was repeated four times. A control was run on the extraction medium (Rhum de
Luxe).
RESULTS
Feeding
At least five species of bamboo are common at the research site. One of these,
C.
cf
uiguieri,
was the principal food
of
golden bamboo lemurs and the only bamboo
species they were observed to eat. They ate shoots, leaf bases, pith, and the viny
part of this bamboo. Greater bamboo lemurs ate the mature culms (stalks) of the
same bamboo species while gentle bamboo lemurs ate the leaf bases and blades of
a related species of bamboo,
C.
perrieri.
Ninety-one percent of the golden bamboo lemurs’ feeding minutes were spent
eating bamboo, while the remainder was spent feeding on one unknown palm fruit
and one unknown
Melastoma
fruit. Feeding began shortly after dawn (0530) and
continued for an average of
30
minutes (n
=
45, range 20-50). These feeding bouts
were followed by rest periods of 30 to
40
minutes, which were followed by another
feeding bout and rest period. Feeding and resting alternated until an extended
rest
period began at 1000 and lasted until 1500. After 1500, feeding and resting periods
continued to alternate until dark at 1745.
From June to August, bamboo shoots are rare and the golden bamboo lemurs
spent 60%
of
their feeding time eating leaf bases and the inside of the viny part of
C.
cf
viguieri.
By
December, new shoots of
C.
cf
uiguieri
were again common, and
the golden bamboo lemurs spent
75%
of their feeding time eating shoots with the
remainder spent eating bamboo leaf bases and the pith from the viny part of
C.
cf
viguieri
as well as the palm and
Melastoma
fruit.
The shoots, leaf bases, and pith of
C.
cf
viguieri
all contained cyanide, with the
pith containing high levels. Each of the more than 200 samples tested turned the
test paper dark blue within the first 30 seconds. Leaf blades and culms of
C.
cf
viguieri
tested negative for cyanide as did all plant parts of
C.
perrieri.
Testing
at
various times during a three-year period (see Collection of Plant Material) indi-
cated no seasonal variation in the production of cyanide.
The cyanide content of the extracted sample was 15 mg HCN per 100 g fresh
weight. The extraction medium (Rhum de Luxe) contained no cyanide.
Body Weight
The only wild-caught
H.
griseus
weighed 770 g, whereas the average weight of
five captive
H.
griseus
at
the Duke Primate Center was 936 g (Table
I).
The
average weight of three
H.
aureus
captured during May 1987 was 1,560 g (Table
I).
One of the adult males was captured again in October 1988 and had gained 120
g.
The only weight for adult
H.
simus
is 2,365 g for one adult male [Meier et al.,
19871.
DISCUSSION
Plants that release more than 10-20 mg HCN per 100 g of fresh plant tissue
are considered potentially dangerous to livestock [Moran, 1954; Kingsbury, 19641.
Therefore, the amount of cyanide
(15
mgilO0 g fresh weight) found in the soft
122
/
Glander
et
al.
TABLE
I.
Lemur
weights
Species
Hapalemur aureus
H.
aureus
H. griseus
H.
griseus"
H. griseus"
H.
griseus"
H.
griseus"
H. aureus'
H. aureush
Sex
F
M
M
M
M
M
F
M
M
Weight
(g)
1,500
1,640
770
1,226
94
1
1,022
1,167
1,540
1,660
"Captive animals held at the Duke Primate Center.
hSame individual captured in
May
and October
1988.
stems and growing tips of
C.
cf
uiguieri
would be dangerous to most mammals, but
was not harmful to the golden bamboo lemurs.
Free hydrogen cyanide is readily absorbed by most animals and is highly toxic
[Poulton, 19831. An oral lethal dose of HCN for the mouse is 3.7 mgikg; dog,
4.0
mgikg; cat, 2.0 mgikg; rat,
10
mgikg [Christensen
&
Fairchild, 19761. The lethal
dose for sheep is 2.0 mgikg body weight [Harborne, 19821. The oral lethal dose of
hydrogen cyanide for humans is 0.5-3.5 mgikg body weight or about 50-250 mg
for a typical adult human [Montgomery, 1969; Christensen
&
Fairchild, 19761.
Assuming that an adult golden bamboo lemur
(1.5
kg) is proportionately as
sensitive to cyanide as the animals above, an average (excluding humans) lethal
dose of 6.5 mg would be found in
43
g of bamboo eaten. The golden bamboo lemurs
ate about
500
g of bamboo per day, or about 12 times the lethal dosage of cyanide
daily. This undoubtedly represents a low estimate because of the inefficiency of the
field extraction method. Much larger amounts of cyanide (90-800 mg HCN per
100
g fresh weight) have been reported from other bamboo species
[
Schwarzmaier,
1977; Bagchi
&
Ganguli, 19431.
We do not know how golden bamboo lemurs avoid cyanide poisoning, but there
are
several possibilities. For most animals, the major mechanism of cyanide detox-
ification involves conversion of cyanide to thiocyanate [Montgomery, 1979
I.
This
process requires the enzyme rhodanese, which is present in most animal tissues,
plus amino acids such as methionine or cysteine to convert cyanide to thiocyanate.
Based on nutritional data for other bamboo species, sulfur-containing amino acids
are relatively low ISchaller et al., 19851. Thus, this method of detoxification seems
unlikely for the golden bamboo lemur.
Another method of detoxification is to keep the stomach acidic, since the
re-
lease of cyanide is inhibited by low pH. For humans and other monogastric ani-
mals, acidic stomach conditions prevent the formation of cyanide, Both the greater
bamboo lemur (which eats mature culms) and the gentle bamboo lemur (which eats
leaf bases) have unspecialized stomachs [Hill, 1953
I
and short, globular ceca that
lack apical narrowing and are the simplest of all the lemurs [Tattersall, 19821. This
simplicity in their stomachs and ceca is surprising given their extremely fibrous
diet, but resembles the digestive system of the panda [Schaller et al., 19851. The
digestive system of the golden bamboo lemur is unstudied.
Bamboo is
a
plentiful resource, but relatively few mammals and certainly no
other prosimians utilize it for food. Probably the best-known herbivore that feeds
almost exclusively on bamboo is the giant panda
(AiLuropoda melanoleuca)
Lemur
Niche
Separation
I
123
[Schaller et al., 19851. During some portions of the year, the giant panda eats
shoots of the genus
Fargesia
as
an exclusive food, but not those of the genus
Sinarundinaria,
which is common in the same area [Schaller et al., 19851. These
shoots have considerable total fiber (76-90%? and are low in nutrients, especially
the sulfur-containing amino acid methionine [Schaller
et
al., 1985; Dierenfeld et
al., 1982; Dierenfeld,
1981
I,
but apparently were not tested for cyanide.
Other mammals utilizing this abundant food resource are two Southeast Asian
rodents
(Hapalomys longicaudatus
and
R
hizomys sinense),
a South American rat
(Dactylomys dactylinus?,
and African monkey
(Cercopithecus mitus kanditi),
and a
South American monkey
(Callicebus moloch)
[Emmons, 1981; Schaller et al., 1985;
Wright,
1985
I.
The gentle bamboo lemur
(H. grzseus)
can be found throughout most of the
eastern rain forest, but the golden bamboo lemur
(H.
aureus)
is among the rarest
primates of Madagascar and only found in the southeastern rainforest. Golden
bamboo lemurs’ geographical distribution is closely linked to that of bamboo and
may be limited to those areas of Madagascar where cyanogenic bamboo occurs.
The panda-like bamboo lemurs have exploited an open niche and appear to
have specialized to the extent of dividing up the niche based on the chemical and
nutrient content of the bamboo. Niche partitioning at this level may be more
common among primates than has been formerly appreciated. Ganzhorn [1988
1
suggested that food selection based on primary and secondary plant chemicals is an
important factor in the mechanisms permitting several primate species to be sym-
patric. Additional studies of the three bamboo-eating lemurs of Madagascar are
presently underway to further define this dynamic plant-primate interaction.
CONCLUSIONS
Madagascar.
as a principal component of their diet.
1)
Three species of bamboo-eating lemurs are sympatric in the rain forests of
2) These three species of lemurs utilize the same habitat, and all eat bamboo
3)
The three species select different parts of the bamboo.
4)
The diet of the golden bamboo lemur contained at least 15 mg of cyanide per
5)
Based on a daily diet of
500
g of bamboo, the golden bamboo lemur ingested
6) The mechanism by which the golden bamboo lemurs avoid cyanide poison-
100 grams of fresh weight bamboo.
about 12 times the lethal dose of cyanide each day.
ing is unknown.
ACKNOWLEDGMENTS
This study was supported by the Sub-Saharan African Cooperative Science
Program of the National Science Foundation (NSF INT-86022861, Wildlife Con-
servation International, World Wildlife Fund-US Primate Program, Duke Uni-
versity Research Council, National Geographic Society, Chicago Zoological Soci-
ety, and the Dourocouli Foundation. We thank the Ministere de la Production
Animale (Elevage
et
Ptiche)
et
des Eaux et Fortits, Department of Water and
Forests of Madagascar, and the Missouri Botanical Garden for their cooperation
and help. Thanks to George Schatz, Porter Lowry
11,
Roan McNab, Anita Brinker,
David Meyers, Patrick Daniels, Joe Macedonia, and our guides, Emile Rajery,
Loret Rasabo, and Pierre Talata of Ambatolahy without whose assistance the
successful completion of this study would have been difficult. Our special thanks to
Tom Soderstrom, David Edelman, and Emmet Judziewicz for identifying the bam-
boo. This is paper no.
448
from the Duke University Primate Center.
124
I
Glander
et
al.
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... These are the bamboo lemurs (Hapalemur spp. and Prolemur simus) (Ballhorn et al., 2009;Eronen et al., 2017;Glander et al., 1989) and, most particularly, the only two exclusively herbivorous Carnivora taxa, i.e. the giant panda (Ailuropoda melanoleuca) (Hansen et al., 2009;Nie et al., 2015;Schaller et al., 1989) and the red panda (Ailurus fulgens) (Panthi et al., 2012;Wei et al., 1995;Wei et al., 1999;Zhang et al., 2009). ...
... In vertebrates, the common mechanism to detoxify cyanide is its conversion to thiocyanate via the enzyme rhodanase (Nahrstedt, 1985) which is also the main mechanism described for pandas (Huang et al., 2016;Zhu et al., 2018). Additionally, a low gastric pH can prevent the release of cyanide from plant material (Glander et al., 1989). Bamboo lemurs (Hapalemur spp. ...
... Bamboo lemurs (Hapalemur spp. and Prolemur simus) ingest high amounts of cyanide (up to 12 times the lethal dose for rodents) but seem to be adapted through mechanisms still to be clarified (Glander et al., 1989). Whether the guinea pigs were capable of detoxifying cyanide by either of these mechanisms remains to be elucidated. ...
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Field and Laboratory Methods in Primatology is a manual for students and researchers studying wild primates. Technological advances allow fieldworkers to collect a wide range of data, store samples for later analysis, and collect information remotely. These methods open up opportunities to gain new insights on previously studied populations and are the means of collecting data on species that have, until now, been difficult to study. However, information on the practicalities of using such methodology in the field has largely been lacking. Here, in this indispensable reference, experienced fieldworkers provide the first comprehensive guide to the wide variety of techniques available for the study of wild primates. Covering everything from pre-trip planning to laboratory analysis of endocrine and genetic samples, packed full of tips and emphasising practicalities and ethics throughout, it is a must-have for all field primatologists and others studying free-ranging animals.
Chapter
Field and Laboratory Methods in Primatology is a manual for students and researchers studying wild primates. Technological advances allow fieldworkers to collect a wide range of data, store samples for later analysis, and collect information remotely. These methods open up opportunities to gain new insights on previously studied populations and are the means of collecting data on species that have, until now, been difficult to study. However, information on the practicalities of using such methodology in the field has largely been lacking. Here, in this indispensable reference, experienced fieldworkers provide the first comprehensive guide to the wide variety of techniques available for the study of wild primates. Covering everything from pre-trip planning to laboratory analysis of endocrine and genetic samples, packed full of tips and emphasising practicalities and ethics throughout, it is a must-have for all field primatologists and others studying free-ranging animals.
Chapter
Field and Laboratory Methods in Primatology is a manual for students and researchers studying wild primates. Technological advances allow fieldworkers to collect a wide range of data, store samples for later analysis, and collect information remotely. These methods open up opportunities to gain new insights on previously studied populations and are the means of collecting data on species that have, until now, been difficult to study. However, information on the practicalities of using such methodology in the field has largely been lacking. Here, in this indispensable reference, experienced fieldworkers provide the first comprehensive guide to the wide variety of techniques available for the study of wild primates. Covering everything from pre-trip planning to laboratory analysis of endocrine and genetic samples, packed full of tips and emphasising practicalities and ethics throughout, it is a must-have for all field primatologists and others studying free-ranging animals.
Chapter
Field and Laboratory Methods in Primatology is a manual for students and researchers studying wild primates. Technological advances allow fieldworkers to collect a wide range of data, store samples for later analysis, and collect information remotely. These methods open up opportunities to gain new insights on previously studied populations and are the means of collecting data on species that have, until now, been difficult to study. However, information on the practicalities of using such methodology in the field has largely been lacking. Here, in this indispensable reference, experienced fieldworkers provide the first comprehensive guide to the wide variety of techniques available for the study of wild primates. Covering everything from pre-trip planning to laboratory analysis of endocrine and genetic samples, packed full of tips and emphasising practicalities and ethics throughout, it is a must-have for all field primatologists and others studying free-ranging animals.
Chapter
Field and Laboratory Methods in Primatology is a manual for students and researchers studying wild primates. Technological advances allow fieldworkers to collect a wide range of data, store samples for later analysis, and collect information remotely. These methods open up opportunities to gain new insights on previously studied populations and are the means of collecting data on species that have, until now, been difficult to study. However, information on the practicalities of using such methodology in the field has largely been lacking. Here, in this indispensable reference, experienced fieldworkers provide the first comprehensive guide to the wide variety of techniques available for the study of wild primates. Covering everything from pre-trip planning to laboratory analysis of endocrine and genetic samples, packed full of tips and emphasising practicalities and ethics throughout, it is a must-have for all field primatologists and others studying free-ranging animals.
Chapter
Field and Laboratory Methods in Primatology is a manual for students and researchers studying wild primates. Technological advances allow fieldworkers to collect a wide range of data, store samples for later analysis, and collect information remotely. These methods open up opportunities to gain new insights on previously studied populations and are the means of collecting data on species that have, until now, been difficult to study. However, information on the practicalities of using such methodology in the field has largely been lacking. Here, in this indispensable reference, experienced fieldworkers provide the first comprehensive guide to the wide variety of techniques available for the study of wild primates. Covering everything from pre-trip planning to laboratory analysis of endocrine and genetic samples, packed full of tips and emphasising practicalities and ethics throughout, it is a must-have for all field primatologists and others studying free-ranging animals.
Chapter
Field and Laboratory Methods in Primatology is a manual for students and researchers studying wild primates. Technological advances allow fieldworkers to collect a wide range of data, store samples for later analysis, and collect information remotely. These methods open up opportunities to gain new insights on previously studied populations and are the means of collecting data on species that have, until now, been difficult to study. However, information on the practicalities of using such methodology in the field has largely been lacking. Here, in this indispensable reference, experienced fieldworkers provide the first comprehensive guide to the wide variety of techniques available for the study of wild primates. Covering everything from pre-trip planning to laboratory analysis of endocrine and genetic samples, packed full of tips and emphasising practicalities and ethics throughout, it is a must-have for all field primatologists and others studying free-ranging animals.
Article
This edition of the Registry of Toxic Effects of Chemical Substances was published in two volumes to accommodate the increased number of substances and toxicity citations, and to provide indexes to those subfiles containing substances of special interest. Rather than split the text arbitrarily, the authors included the indexes in Volume I and all toxicity data in Volume II. Volume I contains the Introduction and Appendixes and is divided into five chapters. Chapter one contains a complete alphabetical listing of all substance prime names and synonyms in the Registry. The remaining four chapters are indexes to the subfiles of those compounds with toxicity citations in the literature indicating carcinogenic and neoplastigenic, teratogenic, mutagenic, or human toxic effects. The Registry contains 98,993 listings of chemical substances. The 1977 Registry includes listings for over 4,700 new substances in addition to more than 8,000 new toxic dose data lines. This edition also includes citations for those substances being tested under the National Cancer Institute's carcinogenesis bioassay program, as well as references to comprehensive toxicity review articles located in the literature.
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Taxiphyllin (1), l-asparagine, 4-hydroxybenzaldehyde and β-sitosterol are identified as the main extractives of the shoots of Dendrocalamus giganteus and D. hamiltonii. An analysis of the free amino acids is presented and the administration of taxiphyllin-[1-14C] to the inner leaves of a bamboo shoot yielding labelled asparagine described.
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The 1975 Edition of the Registry of Toxic Effects of Chemical Substances, formerly known as the Toxic Substances List is the fourth revision of the list of toxic substances prepared at least annually in accordance with Section 20(a)(6) of the Occupational Safety and Health Act of 1970, Public Law 91-596. The Registry includes the original list published in June 1971 and the offerings of the subsequent publications. The Registry contains 64,300 listings of chemical substances: 16,500 are names of different chemicals with qualifying toxic dose information and 47,800 consist of synonymous names and codes which have been acquired from published literature, cooperating industries, and the American Chemical Society. The revision contains approximately 4,000 new compounds which had not appeared in the 1974 edition. The specific data provided on the toxicity of each substance related to: route of administration, animal species involved, type of exposure reported (e.g. lethal), dose which caused toxic response, and body system affected. The bibliographical sources from which the toxicity data were extracted are all listed. Absence of a substance does not imply that it is innocuous.