A RESOURCE UNIT ON ANIMAL REGENERATION
A THESIS
SUBMITTED TO THE FACULTY OF THE SCHOOL OF EDUCATION
ATLANTA UNIVERSITY, IN PARTIAL FULFILMENT
OF THE REQUIREMENTS FOR THE DEGREE
OF MASTER OF ARTS
BY
BOBBIE L. JOHNSON
SCHOOL OF EDUCATION
ATLANTA UNIVERSITY
ATLANTA, GEORGIA
AUGUST 1962 3 k 9^
DEDICATION
This thesis is affectionately dedicated to my wife, Mrs. Patricia Grace Johnson, for her under¬ standing and encouragement, and to my children -
Michael, Michele and Kassondra,
B.L.J#
ii ACKNOWLEDGMENTS
The author wishes to thank all persons who have made the success of this venture possible* Special words of appreciation, however, must go to Drs, E. K, Weaver and
L* Boyd of the School of Education; Drs* G, E. Riley and
M, L. Reddick of the Biology Department, Atlanta
University; Dr, K, A, Huggins, Director of the National
Science Foundation Academic Tear Institute; Drs* H, C.
McBay and Roy Hunter, Jr, of Morehouse College, and Dr,
Joe Hall, Superintendent of the Dade County (Florida)
Public Schools,
B,L,J,
iii TABLE OF CONTENTS
Page
DEDICATION Ü
ACKNOWLEDGMENTS iii
Chapter I. INTRODUCTION ...... 1 Rationale • 1 Evolution of the Problem • •••...••.•• 3 Contribution to Educational Knowledge...... 3 Statement of the Problem U Purpose of the Study . U Limitation of the Study. U Definition of Terms U Materials ...... •••• 5 Operational Steps 6 Method of Research «•••• •••• 8 Survey of Related Literature. ••••••••• 8 H. THE RESOURCE UNIT 13 Outline . . . * 13 Introductory Comments. •••.••••••••. 13 Orientation for the Teacher* Animal Regeneration 18 Introduction ...... 18 Universality of Regeneration 20 Source of the Cells of the Regenerate. . . . . 23 Extrinsic Factors in Regeneration, 2k Stimulus for Regeneration ...... 2$ Organization of the Regenerate ... 26 Release of Block to Regeneration 28 Regenerative Differences in Closely Related Forms 29 Summary ...... 29 References ...... 31 Specific Treatises and Books with Chapters on Regeneration. ••••••••••••• 31 Articles 32 Protozoa ...... •••• 32 Coelenterates (Hydra and Tubularia). , . . 32
iv V
TABLE OF CONTENTS (CONTINUED)
Chapter Page
Flatwonns (Planaria) • •••••••••. 33 Annelids (Earthworms)* •••••••••• 33 Crustaceans (Crayfish) * 34 Vertebrates ••••* . . . . • 34 Statement on the Objectives...... 37 Introduction 37 Principles of Biology Associated with Regeneration Phenomenon. *••.•••••* 37 Suggested Activities for Securing the Objectives 39 Introduction . • 39 Questions on Regeneration 40 Experiments for Answering Question 1 42 Experiments for Answering Question 2 45 Experiments for Answering Question 3 . . • * • 46 Experiments for Answering Question 4 • • * . • 47 Experiments for Answering Question 5 « • • • • 49 Experiments for Answering Question 6 , * * . * 50 Experiments for Answering Question 7 • • • • • 51 Experiments for Answering Question 8 £2 Experiments for Answering Question 9 • • • • « 53 Experiments for Answering Question 10. .... 54 Experiments for Answering Question 11. . * . . 55 Evaluation of the Experiences from the Activities of the Unit 56 Ability to Accurately Observe and Record Data. 56 Operation Skills Developed •••••..••• 56 Ability to Propose new Experiments for Testing Validity of Other Principles Not Included In This Unit 56 Examination 56 Appendix for the Unit. . 56 Animals Required •«••• 56 Methods for Culturing and Caring for Animals in the laboratory 57 Equipment and Supplies Needed. 58 Non-Specific Items •••.•••. 58
III. SUMMARY AND CONCLUSIONS 59
Recapitulation of Experimental . 59 Summary of Literature. * 60 Resume of Findings 6l Conclusions. 63 Recommendations 63
BIBLIOGRAPHY 64 CHAPTER I
INTRODUCTION
Rationale.—The phenomenon of regeneration (the restoration of lost parts) has attracted the attention of man for many years. Much of this concern resulted from the "closeness" of the phenomenon. That is to say, man could not escape the recognition of regeneration because all around him, and even within himself, he could see its manifestations* such as the healing of wounds and the replacement of finger nails and hair. Too, he was most certainly led to wonder why some animals could replace a lost limb and he could not. How amazed were the oyster fishermen when they found they could not destroy the oyster's natural enemy, the starfish, by tearing off its limb. Each of these limbs, when broken in a certain manner, would form an entirely new starfish.
After two centuries of work and thought, men of science, as well as the laity, are still puzzled by the regenerative powers of some animals.
There is, however, one clear point: the ability to replace lost parts has
decreased with advancing evolution of more complex animals. In other words, animals in the "lower phyla" (large categories for classifying living organisms) show more power of regeneration than those in the "higher phyla."
It is also true that, with increasing age, the ability to repair and
to replace is progressively lost, for repair is a measure of the growth-
energies in an individual. These energies are greatest during the early
1 2 stages of the life span. As age accumulates growth-energies diminish.^"
How are lost nails replaced? What causes a wound to heal such that it is often difficult to point out the original site of the injury? How can a fragment of an animal (like a planarian) give rise to a complete and well organized animal? How is it that man cannot restore a lost
finger or leg when some other animals can? These questions are obvious to man because he sees the sources of the questions. Yet there are num¬
erous instances of the replacement of lost parts going on daily within his body about which he is unaware. The lining of the stomach is being re¬ placed almost constantly, and worn-out blood cells are being restored, as well as replaced.
Beyond the "selfish” aspects of the phenomenon of regeneration is its
tremendous biological importance. What is the source of the cells which
will form the regenerated part? Why is it that animals so closely related
as frogs and salamanders show such different regenerative abilities? Why
does regeneration occur more readily only in the young of some species,
and in both young and old of other species? What triggers the regenerative
process itself — is it the marshaling of the necessary cells and synthetic
processes with the ability to duplicate the organization already observed
prior to injury?
It would appear, therefore, that a unit of study on this topic will
be most rewarding to a pupil in gaining some insight into a problem so
basic and so close to himself. The teacher will be in a position to, in a
single unit, touch upon several key biological principles - cell death,
-
L. J. Milne and M. J. Milne, The Biotic World and Man (2d. ed,; Englewood Cliffs, New Jersey: Prentice-Hall, Inc., 19^8),p. 272. 3
cell renewal, tissue organization. Furthermore, it will be possible to observe the effects of environmental factors (temperatures, etc.) on the
regenerative process.
Evolution of the Problem.—This problem grew out of a series of ex¬ periments on animal regeneration conducted by the writer in a course in
Experimental Biology at Atlanta University. The impact of the phenomenon
of regeneration itself, as well as the numerous biological principles in¬
herent in the phenomenon, led the writer to feel that such studies would
provide needed impetus to high school biology courses. The pupil would
undoubtedly be fascinated by direct observations on regenerating systems
to the extent of wanting to raise many important questions about the
process. Several short-term experiments could then be set up to provide
answers to many of the questions. The writer believes that as teacher of
such a group, he could then lead them to a greater understanding of key
issues in biology as well as to an appreciation of the tasks involved in
adequately interpreting the results of the experiments.
Contribution to Educational Knowledge.—The problem to be explored
by the writer will contribute greatly to the pupils' knowledge about some
of the mysterious phenomena happening around themj phenomena which they
can see and marvel at. Too, the pupils will be made more cognizant of
many of the unseen events going on constantly within their bodies. Such
events which they take for granted pose baffling problems for biologists
and would offer significant clues to problems of life itself if they could
be solved. Hence, a practical contribution can be made: the pupils come
to appreciate the tediousness and difficulty involved in the interpretation
of scientific data as well as the setting up of suitable experiments to U answer important questions»
Statement of the Problem»—The problem here is to identify certain phenomena associated with animal regeneration and then to utilize these as the bases for preparation of a resource unit on regeneration to be used in teaching high school biology. Such a resource unit will include data on the kinds of animals best suited for short-term experiments on regenera¬ tion and on specific experiments designed to answer key questions.
Purpose of the Study»—The ultimate objective of this study is pre¬ paring a prospectus for teaching certain selected principles in a course in high school biology. The prospectus will be one that is "alive" and challenges the imagination and thinking of the student, while fascinating him. Furthermore, the student will become directly involved in what he is doing and, in all probability, will exhibit more intellectual curiosity about what is happening, especially since he will be able to design means of directing the course of his experiments. The teacher can, therefore,
take advantage of this kind of pupil exuberance to get over many more basic principles of biology.
Limitation of the Study,—The material presented in this study will
touch directly upon the gross aspects of the regenerative process. In
order to gain a deeper insight into the minute details of the phenomenon,
inferences will have to be drawn from the gross observations. The study is
further limited in that the entire animal kingdom cannot be dealt with,
hence only certain phyla and/or classes will be studied.
Definition of Terms.—Some of the significant descriptive terms to be
used in the study are defined below:
"Resource Unit" - A systematic and comprehensive survey, analysis, and 5 organization of the possible resources (e,g., problems, issues, activities, bibliographies, and the like) which a teacher might utilize in planning,
developing, and evaluating a learning unit.^
'•Regeneration" - The repair by growth and differentiation of damage 2 suffered by an organism past the phase of primordial development,
"Mitosis" - A form of cell division characterized by ,., exact
chromosome duplication,
"Metabolism" - A group of life-sustaining processes including prin¬
cipally nutrition, production of energy (respiration), and synthesis of more protoplasm, 3 "Growth" - Increase in cell size or in cell number,
"Vertebrates" - Animals possessing a cranium and a spinal column of
vetebrae which form the chief skeletal axis of the body. It "Invetebrates" - Animals without a spinal column of vetebrae.
Materials,—The materials to be used in the experiments proposed in
this study are the following:
(l) Invetebrates
(a) Stentor - a protozoan
1 Harold Alberty, "How to Make A Resource Unit" (unpublished bulletin, College of Education, The Ohio State University Press, 19WJ), p. 5« 2 P. Weiss, Principles of Development (New York: Henry Holt Company, 1939), p. U58. 3 P. B, Weisz, The Science of Biology (New York: McGraw-Hill Book Company, 1959), p. 583. k C, P, Hickman, Integrated Principles of Zoology (St, Louis, Missouri: C, V, Mosby Company, 1955), p. 3$lu 6
(b) Hydra - a freshwater coelenterate
(c) Tubularia - a marine coelenterate
(d) Planaria - a free-living fiatworm
(e) Earthworm. - an annelid worm
(f) Leech - an annelid worm
(g) Crayfish - a crustacean (artfchropod)
(h) Starfish - an echinoderm
(2) Vertebrates
(a) Goldfish - a bony fish
(b) Frogs (tadpoles and adults) - tailless amphibians
(c) Salamanders (adults) - tailed amphibians
(d) Mouse - a mammal
Operational Steps.—-The operational steps in this study will be as follows:
1. Several questions will be posed to test certain key hypotheses
about animal regeneration.
2. Suitable animals, representing "lower" and "higher" groups, will
be selected. The suitability of the animals will be based on
their known ability to show rather rapid regenerative powers.
(a) These animals will then be categorized as follows:
(1) Ability of a fragment to give rise to a whole animal
(2) Ability of an animal to restore that part only
3. Experiments will be carried out to establish the rate of re¬
generation under controlled conditions of temperature, growth
medium constituents.
U. In order to show the significance of these factors in growing
systems, experiments will next be conducted wherein temperature 7
and changes in growth medium will be varied. The information
obtained here will be correlated with the "controls" of number
three.
5>. Each of the above experiments will be prefaced with particular
questions to be answered by the results of the experiments.
6. A Resource Unit will be prepared centered around the following
biological principles :
Regeneration is almost universal among living thingsj from the simple to the more complex animals, the abilities to regenerate lost parts and to reproduce asexually, fall off, gradually and independently, as the body becomes more specialized.
Growth and repair are fundamental activities for all protoplasm.
Growth and development in organisms is essentially a cellular phenomenon, a direct result of mitotic cell division.
All cells arise through the division of previous cells (or protoplasm), back to the primitive ancestral cell (or pro¬ toplasm). Cell division is the essential mechanism of reproduction, of heredity, and to a large extent, of organic evolution.
Cells are organized into tissues, tissues into organs and organs into systems, the better to carry on the functions of complex organisms.
The protoplasm of a cell carries on continuously all the general processes of any living bodyj the processes concerned in the growth and repair or upbuilding of protoplasm (anabolism) and the processes concerned with the breaking down of protoplasm and elimination of wastes from the cell (catabolism). The sum of all these chemical and physical processes is metabolism.
Prom the lower to the higher forms of life, there is an in¬ creasing complexity of structure, and this is accompanied by a progressive increase in division of labor. In all organisms, the higher the organization the greater the degree of differ¬ entiation and division of labor and of the dependency of one part upon another.
Cell division is the essential mechanism of reproduction of heredity, and to a large extent, of organic evolution. 8
The environment acts upon living things, and living things act upon their environment. Since the environment of living things changes continually, these creatures are continually engaged in a struggle with their environment.
The range of temperature for life activities is very narrow as compared with the range of possible temperatures. There is a minimum temperature below which, and a maximum temperature above which, no life processes are carried on. The temperature range for life processes is from many degrees below 0° C, to nearly the boiling point of water.
Adult organisms that differ greatly from one another but which show fundamental similarities in embryological development, have originated from similar ancestors.
Animals resemble each other more and more closely, the farther back we pursue them in embryological development.
Method of Research,--The principle-question-experiment method will be employed in this study. Such a procedure will permit the student to realize at once the principles involved. Then specific questions may be proposed about the principle. The experiment will provide experiences that may be discussed in the light of the principle, with appropriate conclusions to be drawn from the results.
Survey of Related Literature,—The success or failure in the teaching of any subject rests, to a large degree, in the ability of the teacher to make each lesson vitally interesting and worthwhile to the pupil. Suf¬ ficient experimental evidence indicates that if there is abundant present- day life activity in the classroom, the child is more likely to develop a lively interest in the subject field. Success in the promotion of this type of lesson depends largely in the development of certain fundamental
1 M, J, McKibben, "An Analysis of Principles and Activities of Importance for General Biology Courses in High Schools,” Science Education, XXXIX (April, 1955), pp. 187-196, 9 techniques in the learning process.
One of the main objectives of science instruction is to develop in students the habits of thought and action inherent in the so-called methods and attitudes of science. There is a need in science courses for setting classroom situations which will be conducive to the development of the ability to communicate and to participate effectively in group dis- 2 eussions. One of the best ways to secure these objectives is through the medium of the laboratory exercise. The value of problem-solving through laboratory work in the school does not lie in the factual knowledge that may result from it but in the attitudes and habits of reflective thinking it encourages and in the understanding it gives of how the knowledge of science gained by the student from description was attained in the first 3 place. Appropriate criticisms of research can be made more intelligible when one is made aware of underlying principles.
Every school is, in essence and purpose, and experimental school. All of us are trying to improve our teaching. Our effectiveness in this process of improving our instruction depends upon our philosophy, and upon our skills in implementing our philosophy through provision of opportunities for students to learn...In the curriculum there is definite need for the development of procedures and techniques for directing effectively the
_ M. P. Simmons, "A Model Lesson in General Science," Science Education, XXIII (March, 1939), p. 133. 2 J. M. Mason and W. G. Warrington, "An Experiment in Using Current Scientific Articles in Classroom Teaching," Science Education, XXXVIII (October, 19&), p. 299. 3 Science in General Education. Report of the Committee on the Function of Science in General Education (New York: D. Appleton Centüry Co., 1938), p. 317. 10 activities of the learner. The resource unit is the procedure for directing the activities of the learners.^ The unit to be described in this study will provide a directive for students and teachers in high school biology courses with the hope that such a directive will represent a somewhat radical departure from existing methods in such courses.
Most of the experiments presented in present-day science manuals and carried out in science laboratory periods, preclude the possibility of much reflective thinking on the part of the student. Too often the student's job is to follow direction in an effort to achieve accuracy in results he already knows. Too often the subject of the experiment has little or no real interest and motivation for the student, who, as a con¬ sequence, blindly follows the directions without question or understanding.
Under these circumstances it can hardly be expected that the student will 2 develop resourcefulness and facility in employing the scientific nethod.
The study of regeneration phenomenon promises to open many fruitful avenues of approach toward the solution of such basic problems of develop¬ ment as growth, differentiation, morphogenesis, and secondary adjustments 3 between parts and the rest of the organism. The word regeneration is used to refer to the processes by which an animal restores, or tends to restore, any regions which may be removed. At one extreme an adult mammal which has suffered the loss of a small part, such as a finger, or a larger part,
_ Edward K. Weaver, "How to Make a Resource Unit" (unpublished com¬ pilation, The State Teachers College, Montgomery, Alabama, Summer Session, 1946), p. 2. 2 R. W. Burnett, "Vitalizing the Laboratory to Encourage Reflective Thinking," Science Education, XXIII (March, 1939), p. 138. 3 Weiss, op. cit., p. 478. 11 such as a finger, or a larger part, such as a limb, can do more in the way of regeneration than merely repair the wound and close the cut surface.
At the other extreme a very small part of the normal body of a coelen- terate, a flat worm, or a starfish, can restore the whole large region which is missing and become a complete individual. There are all grades 1 in between these two extremes.
On the whole, one gains the impression that regenerative capacity tends to vary inversely as the scale of organization. Generally speaking, the percentage of good regenerators is lower among the higher forms than among the lower, more simply organized forms. But when it comes to par¬ ticulars, there is not a single group of animals whose regenerative capacity could be safely predicted from its position on the evolutionary scale alone. There are lowly forms with practically no regenerative capacity, for example, the ctenophores and rotifers; while, on the other hand, some higher forms, such as the crustaceans and amphibians, re¬ generate amazingly well. Moreover, closely related forms, such as the earthworms and the leeches, or the tailed and tailless amphibians, may often represent diametrical extremes with regard to regenerative capacity: the earthworm and urodele amphibians are excellent regenerators, while 2 the leeches and anurans are among the poorest,
A basic problem in biology is that concerning the stimulus for re¬ activating developmental processes after development has been, as it were, halted. In other words, is an adult organism a fixed structure, incapable
1 C, H. Waddington, Principles of Qribryology (London: George Allen and Unwin, 1957), p. 302, 2 Weiss, op, cit,, p, U59. of change, or does it still show some of the labile organization so 1 characteristic of the egg? The act of regeneration clearly indicates that an adult organism is capable of change. The process of regeneration gives rise to some new problems in development. One of them concerns the origin of the cells which give rise to the regenerating structure. We might visualize either that the new structure comes from the old cells by migration or cell division or that there may be reserve cells, embryonic in nature, which are undifferentiated and which give rise to the new structure,^
1 L. B, Barth, Embryology (New York: Henry Holt and Company, 1953), P. 327. CHAPTER II
THE RESOURCE UNIT
This chapter presents the resource unit on "Regeneration" for High
School Biology Teachers, The basic factual materials for this unit were primarily derived from a series of "experimental" studies carried on in the Biology Department of the Atlanta University Schools of Arts and
Sciences during the summer sessions of 19f>7 and I960, All other materials presented in the resource unit were gathered through copious reading and reference work in the Trevor Arnett Library, Atlanta University, during the first and second semesters of the 1961-1962 academic year, A brief outline of the contents of the unit is provided below, together with certain introductory and other explanatory comments.
Since all of us are trying to improve our teaching, our effective¬ ness in this process will depend upon our philosophy and upon our skills in implementing our philosophy through the provision of opportunities for students to learn. The resource unit is the procedure for directing the activities of the learners. The unit to be described will provide a directive for teachers in high school biology courses, with the hope that such a directive will represent a somewhat radical departure from existing methods in such courses.
Outline
I, Introductory Comments
II# Orientation for the Teachers Animal Regeneration
13 lli
A. Introduction
B. Universality of Regeneration
C* Source of the Cells of the Regenerate
D. Extrinsic Factors in Regeneration
E. Stimulus for Regeneration
F. Organization of the Regenerate
G. Release of Block to Regeneration
H. Regenerative Differences in Closely Related Forms
I. Summary
J. References
1. Specific Treatises and Books with Chapters on Regeneration
2. Articles
a. Protozoa
b. Coelenterates
c. Flatworms
d. Annelids
e. Crustaceans
f. Vertebrates
III, Statements on the Objectives
A, Introduction
B, Principles of Biology Associated with Regeneration Phenomena
TV, Suggested Activities for Securing the Objectives
A, Introduction
B, Questions on Regeneration
C, Experiments for Answering the Questions
D, Evaluation of the Experiences from the Activities of the Unit 15
1. Ability to Accurately Observe and Record Data
2. Operation Skills Developed
3. Ability to Propose New Experiments for Testing Validity
of Other Principles Not Included in This Unit
U. Examination
V* Appendix for the Unit
A. Animals Required
B. Methods for Culturing and Caring for Animals in the
Laboratory
C* Equipment and Supplies Needed
D. Non-Specific Items
Introductory Comments
Any scientific study or activity, involving phenomena which not only
“strike one's fancy" but which are of tremendous scientific importance as well, is likely to be most rewarding to the participant. A student in a biology course who can, by active and well-planned experimentation, come to grips with a phenomenon so close to him as regeneration of damaged or lost body parts, will acquire immeasurable knowledge about some key biological principles. This is eo because phenomena associated with re¬ generation itself touch upon and, to be sure, are part of several funda¬ mental principles of biology in general. For example, it is well known that adult frogs and salamanders (both are amphibians) show different re¬ generative abilities. When a salamander's limb is amputated, a new one forms. This does not occur normally when a frog's leg is amputated. Hoi*, ever, both frog and salamander larvae show remarkable capacities for re¬ generation. Such a simple observation on such closely related animals as 16
frogs and salamanders clearly illustrates the principle that animals
resemble each other more and more closely the farther back we pursue them
in embryological development,
A salamander's limb may be de-boned (that is, all bones are removed, by surgical procedures) and amputated. The amputated surface will heal
and, in time, a new limb regenerates, complete with bony partsJJ Where did
the new cells come from for the "boned” new limb which regenerated from
a "de-boned" stump? First of all, one of the basic principles of biology
is that all cells arise through the division of previous cells. Since this
is true, the above observation on limb regeneration suggests that cells
which were originally one thing can become something else; for it is
clear, that some of the remaining cells in the de-boned stump becane , as
it were, triggered into becoming bone cells. This fact leads up to the
principle that growth and development and repair in organisms is essentially
a cellular phenomenon, a direct result of mitotic cell division.
Finally, a tiny fragment of an animal like a planarian can give rise
to a whole organism; however, if a dog's tail is cut off at any stage in
its life, no new tail forms, A planarian is a flatworm, an irorertebrate
animal; a dog is a mammal, a vertebrate. Such an occurrence fits another
of the generalizations in biology: While regeneration is almost universal
among living things, from the simple to the more complex animals, re¬
generative abilities fall off as the body becomes more specialized.
It would appear, therefore, that a resource unit on animal regeneration
will provide many experiences that can be quite beneficial and rewarding
to a pupil. The usefulness of such an approach to teaching biological
principles gains support from the following: There appears to be a growing 17 concern over the country about the development of scientific attitudes and the abilities of critical thinking or problem solving ...Some of the needed studies recommended...in the area of problem solving ares a. Tech¬ niques of setting up problems to be solved in a unit of workj b. Learning experiences designed to help pupils identify problems; c. Processes in¬ volved in proposing and testing hypotheses; d. Learning experiences to help pupils interpret evidence; e. Learning experiences designed to 1 help pupils draw conclusions from data.
From such a unit proposed in this thesis, the student can gain in¬ sight into a problem so basic to biology and yet so close to himself. Too, he will obtain satisfaction from verifying accepted principles for himself, as a result of carrying out the proposed experiments. In this way the student visualizes the facts instead of just memorizing than. The teacher will be in a position to take advantage of such a learning situation and stress several key biological principles from a single ’'living" unit of study.
Prior to the use of the suggested activities in this unit, it would be well for the teacher to make a survey of the pupils' awareness of such happenings in nature, including those within their own bodies. That is to say, he (the teacher) might askî Have you ever heard of an animal losing a part of its body and having it restored? What kinds of animals seem to possess this ability? Have you ever wondered why it is that a human being does not restore an amputated limb? Are there any parts of your body which are being repaired or restored? Do you look upon this
-
E. S. Oboura, L. H. Darnell, G. Davis, and E. K. Weaver, "Fifth Annual Review of Research in Science Teaching," Science Education, XLI (December, 19S7), p. UOU. 18 restoration activity as being a simple or complex phenomenon? What are
some of the problems involved in such activity? How would you go about trying to determine if all kinds of animals, simple and complex ones, have the ability to restore lost or repair damaged parts?
Following this brief survey, the teacher can go into details about certain aspects of regeneration (as selected from the Orientation to the
Teacher); after which the class can be divided into small groups to carry out selected experiments designed to stress basic principles of biology.
The students should be allowed to draw conclusions from their observations,
discuss them with their other classmates, and all of these activities are
to be coordinated by the teacher.
Orientation for the Teacher: Animal Regeneration
Introduction
One of the significant qualities of many animals, during their embry-
ogeny , is that of plasticity. That is to say, certain parts of the embryo
are not irreversibly fixed, but can be altered to such an extent that these
parts can become something entirely different from their expected fate. In
time, however, as the embryo ages, there is a progressive decrease in the
“lability” and an increase in the "stability” of embryonic parts. In
other words, the older the developing organism becomes, the more fixed
(or determined) will be its many parts.
Once the organism has reached an advanced stage in its life history
(adulthood, for example), can those "fixed” areas repair or restore them¬
selves if damaged or lost? Will there always be a reservoir of the "labile"
embryonic cells on hand to re-form any losses suffered by the adult? Gan
the "stable" cells of the adult be stimulated to become "labile" cells?
How "fixed" is an adult animal? Is it so rigidly fixed or stable that it 19 cannot exhibit any degree of plasticity or lability?
The evidence is clear that the parts of adult animals are not rigidly determined. They can repair and restore damaged or lost parts in numerous instances. This fact is so obvious from our general knowledge about our own body as to be taken for granted. We cut ourselves and the wound heals.
We lose a nail and a new one develops. We break a leg bone and it repairs itself. We are told that billions of our red blood cells become worn out daily, and yet we do not suffer (under normal conditions) any ill effect
since these cells are replaced.
On the other hand we are equally aware that when we cut off the tail of our pet dog or cat, no new tail forms. Should one of our legs or arms be amputated, there is no restoration of the lost part. Yet when the
fisherman cuts his bait (an earthworm) into two parts, he finds that after
a time there are two earthworms. The chief enemy of the oyster industry,
the starfish, if severed into five parts, may after a while form an entirely
whole organism from each of the five fragments. Just as noticeable is
the ability of the little boy's pet salamander to restore a missing limb,
or his tadpole to replace part of a severed tail.
These and many other rather obvious occurrences are rightfully puzzling
to any man. Yet, to the student of life phenomena these incidences of
regeneration present new problems of animal development. Some of these
problems are as follows:
1. Is there any direct correlation between the ability of an
organism to regenerate and the position it occupies on the evo¬
lutionary scale of animal organization? If so, are there both
quantitative and qualitative manifestations in this capacity 20
to replace or repair lost or damaged parts?
2. What is the source of the cellular materials involved in re¬
storing lost cellular areas? Do they rise anew or are they
produced by thetransformation of cells already present?
3, Can the capacity for regeneration be controlled by extrinsic
forces, or is it an expression of intrinsic factors only?
U* What initiates a re-awakening of the ability of a seemingly
well determined and stable organism to exhibit, once again,
capacities usually reserved for the embryo?
Does the regenerated part acquire the same, in all details,
organization associated with the normal structure?
6, How valid is the statement that the "ability of organisms to
regenerate is not lost but that certain conditions of healing
may create a block to cellular activities in the area of the
severed structure?" If such a block is removed, may regeneration
occur in those forms which normally do not regenerate?
7* May closely related organisms show strikingly different capa¬
cities for regeneration?
The information to follow attempts to relate the evidence from experimental morphology which seeks to provide clearer insight into these problems if not answers.
Universality of Regeneration
Regeneration appears to be a universal phenomenon within the animal kingdom. Although present in nearly all animal groups, the ability to regenerate missing portions of the body differs in scope and its course within specific groups, Trembley (around 171*0) first decribed this act (regeneration) in the fresh water coelenterate, Hydra, One of these 21 organisms may be cut in two or more parts and each will reconstitute itself into a new and complete individual, although somewhat diminished in size*
Numerous other invertebrates form (including other coelenterates) are known in which major portions of the body can be repeatedly restored after loss.
Among such organisms are representative protozoa, flatworms, nemerteans, annelids, arthropods, echinoderms, and tunicates. In most of these a fragment measuring only a small fraction of the original body can become a complete individual.
Experiments with Stentor, a protozoan, have shown that regeneration
can occur in fragments containing portions of the nucleus. Even here, how¬
ever, there must be a minimal volume relationship between the nucleus and the cytoplasm in order to get regeneration. In coelenterates like Hydra,
small sections of the body, comprising as little as 1/200 part of the original individual, can regenerate a complete whole, Flatworms such as
Planaria show remarkable regenerative abilities. If one of these animals
is transversely cut into halve», within a period of a few days visible
signs of regeneration are already manifest. This "bud’1 grows steadily and
soon replaces the lost part of the body. That is to say, a new fore-part
is formed on the caudal half, and a new hind-part on the anterior half,
Whether the cut is made in the middle, or in the front or hind part of the worm, the result is always the same*
Segmented worms (annelids) like the earthworms show a somewhat similar
type of regenerative capacity. It has been found in such animals that the
regeneration-bud which develops at a front edge will always differentiate
into a fixed number of segments, which agree with those found in the rostral
end of the body, the Hheadw of a normal worm. The number of segments in
the regenerate is entirely independent of the place of the cut. In arth¬
ropods regeneration is confined to the renewal of lost appendages. For 22 example, in most crustaceans the limbs may regenerate at any stage of de¬ velopment including the adult; however, in insects, limb restoration occurs only in the larval stages, and the regenerated limb usually does not reach the size of a normal limb. Among the echinoderms, the starfishes can regenerate arms and parts of the central disc. The arms appear to be lost rather frequently in the natural environment of these animals, as individuals regenerating one or more arms are found quite often.
The regenerative capacity of vertebrates is much less extensive than that observed in invertebrates. Even in the most favorable cases, such
ability appears to be confined to the replacement of organs, never entire bodies! Among fishes the restoration of fins and scales has been studied widely. In amphibians regenerative power is high during the larval stages of nearly all forms; and in tailed amphibians, it persists even in the
adult stage. These animals can adequately replace parts removed from eyes, jaws, limbs, gills, tail, and several of the internal organs. While an
adult tailed amphibian like a salamander can replace a lost limb, an adult
tailless form like a frog cannot.
Reptiles (lizards, e.g.) can restore lost tail parts. Even birds and mammals are not without some regenerative abilities. Lost feathers may be
restored in birds; in mammals, the phenomenon of wound healing is quite
obvious, and so is the restoration of a lost nail, deer antlers, bone
repair, and others.
From the information provided above it is quite clear that in some
forms regenerative capacity is enormous, while in others it is almost
neglible. On the whole, one gains the impression that regenerative ability
tends to vary inversely as the scale of organization. That is to say, in 23 general terms, the percentage of good regenerators is lower among the higher forms than among the lower, more simply organized, forms. Never¬ theless, when it comes to particular instances, there is not a single group of animals whose regenerative ability could be safely predicted from its position on the evolutionary scale alone. There are lowly forms with practically no restorative capacity, for example, ctenophores and rotifers; while, on the other hand, some incomparably higher forms, like crustaceans and amphibians, regenerate amazingly well.
Source of the Cells of the Regenerate
When a portion of an organism is removed, there are three sources from which the cells which build up the regenerated part may be derived:
1. The tissues of the stump may grow out and form the new organ
or part.
2. The body may contain a reserve of undifferentiated or embryonic
cells which accumulate at the wound and then differentiate into
the tissues of the regenrated part.
3. Some of the already differentiated tissues may lose their
differentiation and assume a more plastic condition from which
they are able to re-differentiate into the specialized tissues
of the regenerate.
Observations from experimental morphology have provided evidence that in animals such as coelenterates, flatworms and some annelid worms, the body contains a supply of reserve cells (called neoblasts) which play a part in regeneration. The stimulus of the wound activities these undif¬ ferentiated cells. In vertebrated animals, however, there is still no full agreement as to the origin of the cells of the regenerate. There appears 2k rather overwhelming evidence that there is a dedifferentiation of the tissues in the near neighborhood of the wound, and that this is by far the most important source of the regenerating cells.
Extrinsic Factors in Regeneration
Extrinsic forces may play significant roles in regeneration. The rate of regeneration is dependent on temperatures, as most biological processes are. Increase of temperature, up to a certain point, accelerates re¬ generation. For example, in Planaria regeneration is scarcely possible at a temperature of 3°G. Of six individuals kept at this temperature only one regenerated a head, and this was defective} the eyes and brain were not fully differentiated after six months. Regeneration was most rapid at 29.7°C.J at this temperature new heads developed in U.6 days. A tem¬ perature of 31.5>°C. was too high, and the heads regenerated after 8,5 days. A temperature of 32°C. proved to be lethal for the animals.
Food, on the other hand, does not affect regeneration very much. Even a fasting animal will regenerate at the expense of its own internal re¬ sources. In such diverse cases as rats regenerating parts of the liver, salamanders regenerating limbs or hydras and planarians regenerating parts of their body, depriving the animals of food does not prevent regeneration and may even accelerate it to a certain extent. If planarians are deprived of food for a long time, they can live by metabolizing constituents of their own bocfcr. The animal, of course, diminishes in size as a consequence.
In this state a planarian can still regenerate. Although the over-all size increases, the missing parts are gradually rebuilt so that a complete, even if small, worm is eventually developed. Although restriction of feeding 25 seems to be favorable for regeneration, if anything, extreme degrees of emaciation by starving prevent regeneration except in organisms such as the planarian which is able to utilize its own body as a source of energy with¬ out deleterious results.
Stimulus for Regeneration
In order for regeneration to take place, certain conditions must be met. First of all there must be a stimulus to provoke the restorative activities. There must be cellular material with an adequate supply of
"embryonic" potencies to produce all the new tissues necessary. There must be organizing factors to make the regenerate appear exactly like the normal structure or organisms.
According to definition, regeneration is the replacement of lost parts. One could expect, therefore, that the loss of some part of the body would be the adequate stimulus to set in motion the mechanism which re¬
stores the part, and thus restores the normal structure of the animal.
This is by no means always the case. If a deep incision is made on the
side of a salamander^ limb, or on the side of the body of an earthworm or
a planarian, a regeneration bud (blastema) may be formed on the out surface.
The bud then proceeds to grow and develop into a mew part, as in ordinary
regeneration. In the case of a limb the new part thus developed will be
the distal part of the limb, from the wound level outward. The development
of the regenerating part proceeds just as if the entire distal part of the
limb were cut off.
In the case of a planarian a lateral incision may cause the develop¬ ment from the wound surface of either a new head or a new tail, or both.
If both a head and a tail are regenerated, the head forms from that part 26 of the wound surface facing posteriorly. This results, of course, in the regenerated head lying more anterior than the regenerating tail. A some¬ what similar reaction is produced by lateral incisions in the earthworm, with the restriction that lateral incisions near the head end of the worm give rise to additional heads, incisions in the middle part of the animal cause the development of both heads and tails, while incisions in the pos¬ terior part of the animal^ body cause the formation of tails only. Another peculiarity, in the case of the earthworms, is that the incision must be deep enough to sever the ventral nerve chain if any regeneration at all is to take place.
Careful study of each of the above mentioned cases will show that the original parts of the animal (heads, tails, limbs) had not been removed, so that the regenerated parts are additional and/or superfluous to the animal. The experiments allow us to draw the conclusion that not the absence of an organ but the presence of a wound is the stimulus for re- generation. Thus it would seem that the capacity for regeneration is not a novel and secondarily acquired adeptness of the organism at meeting later accidents by adequate repairs, but is simply a residue of the original capacities for growth, organization, and differentiation through which the individual was first formed. Hence, the extent of regenerative capacity is limited by the extent to which formative capacities survive the on¬ togenetic phase.
Organization of the Regenerate
Unless the regenerate is structured exactly like the normal structure, one may question its occurrence. In a hydroid, similar to Hydra (Having a base for attachment, a long body, and a set of tentacles which take in 27 food), if we cut it transversely into two parts and follow the development of each of these two parts, we find that one end will form the tentacles and the other end will simply form a base. It is the upper end which forms tentacles; the lower forms a base.
If an animal of simple structure, e.g., a planarian, is cut into two parts transversely, a bud-like out-growth, within a few days, will develop on the cut surface in each half. (This is the so-called regeneration-bud.)
It grows steadily, and replaces the lost part of the body, i.e. a new fore¬ part is formed on the caudal half, and a new hind-part on the anterior half. The position of the cut is of no, or at most of secondary, im¬ portance. Whether it is made in the middle, or in the front or hind part of the worm, the result is always the same. Even if the worm is divided
into several parts by a series of transverse cuts, each of them will re¬
generate a head at its front edge, and a tail at its hind edge.
In earthworms, such as Eisenia foetida. a number of regions with dif¬
ferent powers of regeneration can be distinguished. In the foremost six
segments of the body, a head is regenerated at a front edge, but no re¬
generation takes place at a hind edge. In the succeeding zone of about
eleven segments, heads are formed by both front and hind edges. Next
comes a region, extending to segment fifty-four, in which only tails are
regenerated at both edges. Finally, in the zone hehind segment fifty-
four, a tail is formed by a hind edge, but no regeneration occurs at a
front edge. This earthworm (Eisenia), even in young stages, has 100 seg¬
ments. When the worm is amputated at segment fifty, new segments re¬
generate posteriorly until the total is 100j amputated at eighty, twenty
new segments regenerate posteriorly.
It has been found that in many annelid worms the regeneration-bud 28 which develops at a front edge will always differentiate into a fixed number of segments, which agree with those formed in the rostral end of the body, the ’'head" of a normal worm. The number of segments in the re¬ generate is entirely independent of the place of the cut. This shows that here regeneration does not lead to restitution of the missing part of the body, but produces only a new apical end by an autonomous differentiation.
The same applies to planarians. Here, again, regeneration at a front edge, irrespective of its position, produces only a head, whereas that at a hind edge leads to complete restituttion of the missing parts.
Release 6f Block to Regeneration
Normally the adult frog limb after amputation will heal, but no re¬ generation blastema will form. If, however, one stimulates the amputated surface with salt solutions, the limb forms a blastema. Later the blastema begins to differentiate into the missing limb. Hence, it appears that the usual stimulus of simply cutting through the limb is not suf¬ ficient. Probably in the amputated stump of a frog limb restorative processes are blocked in some way. As already observed, such a block is not associated with an amputated salamander linibj it normally regenerates after amputation.
Failure of a frog limb to regenerate appears to be related to a
failure of the tissues near the cut surface to undergo necessary changes.
The skin merely heals over the wound, and no further action occurs. In
amputated salamander limbs, if the skin is pulled over the cut surface,
no regeneration occurs. That is to say, in an animal in which regeneration
normally occurs, if the wound is covered with skin immediately after
amputation, regeneration is blocked. Such a phenomenon might explain the 29 absence of regeneration in adult frog limbs because it is a normal occur¬ rence for the skin (both outer and inner layers) to close over the wound quite rapidly during the healing process. When salt treatment is applied,
the inner layer of the skin fails to cover the wound, and under such treatment regeneration occurs in the frog limb. Hence, it seems that when both inner and outer layers of the skin close over the wound, no regeneration
takes place; when only the outer layer covers the cut surface, regeneration
ensues. The ability of organisms to restore lost parts does not seem to be lost as such, but conditions of healing arise which present a block to necessary changes in the cells around the cut surface. If such a block
can be alleviated by chemical (or other) treatment (increasing nerve.’
supply), restorative processes take place.
Regenerative Differences in Closely Related Forms
All of these studies on regeneration fail to tell us why the fishes
regenerate much less than amphibians, which, during the course of evolution,
emerged from fish ancestors. Too, no answer is forthcoming to the striking
differences in regenerative capacities between amphibians with tails
(salamanders) and those without (frogs), or the truly segmented worm
(annelids) like earthworms and leeches. By and large, however, when one
looks at the gross picture of regeneration, it would appear that "the
whole is less than the sum of its parts"; for in certain organisms even a
tiny fragment can give rise to a whole and completely organized organism.
The truth is, however, that we have not come to understand fully "the
wholeness of the whole."
Summary
It is apparent, therefore, that the phenomenon of regeneration is 30
■universal among animals. From protozoa to mammals, some kind of res¬ torative processes occur. The quantity of repair or restoration does seem to correlate with the histological complexity of the organism and its position on the evolutionary scale of organization. That is to say, generally speaking, the best regenerators are the simplest organized organisms. On the other hand, the evidence is quite clear that the quan¬ tity and quality of regenerative processes may be controlled by extrinsic factors. References
Specific Treatises and Books with Chapters on Regeneration
Adams, A. E. 1959 Studies in Experimental Zoology (Regeneration,, Experi¬ mental Qnbryology, Endocrinology). Edwards Brothers, Inc,, Ann Arbor, pp. 7-19*
Balinsky, B. I, i960 An Introduction to Bribryology. W. B. Saunders Company, Philadelphia, pp, 472-508.
Barth, L. G. 1953 Embryology, rev, ed, Henry Holt and Company, New York, pp. 327-338*
Barth, L, G, Regeneration: Invertebrates, In: Analysis of Development (B, H. Willier, P, Weiss, and V, Hamburger, eds.), W, B. Saunders Company, pp, 664-672.
Berrill, N, J, 1961 Growth, Development, and Pattern, W, H, Freeman and Company, Inc,, San Francisco, pp. 244-320, 358-1*02. von Bertalanffy, L, 1962. Modern Theories of Development: An Introduction to Theoretical Biology" Harper and Brothers, New York, pp. 168-172."
Bliss, De. I960 Autonomy and regeneration. In: The Physiology of Crustacea, Vol. I (T. H. Waterman, ed,), Academic Press, Inc, New York,
Bodenstein, D, 1953 Regeneration. In: Insect Physiology (K. D, Boeder, ed.), John Wiley and Sons, Inc., New York,
Goss, R. J. 1961 Regeneration of vertebrate appendages. In: Advances in Morphogenesis, Vol. 1 (M. Abercrombie and J. Brachet, eds.), Academic Press, Inc,, New York, pp, 103-149,
Needham, A, E, 1952 Regeneration and Wound-Healing. John Wiley and Sons, Inc., New York.
Needham, J. 1942 Biochemistry and Morphogenesis. Macmillan Company, New York, pp. 430-456,
Nicholas, J. S. 1955 Regeneration: Vertebrates. In: Analysis of De¬ velopment (B. H. Willier, P. Weiss, and V. Hamburger, eds.), W. B. Saunders Company, Philadelphia, pp. 674-693*
Raven, C. P. 1959 An Outline of Developmental Physiology. Pergamon Press, Inc., New York, pp. 166-189*
Thornton, C. S. (ed.) 1959 Regeneration in Vertebrates. University of Chicago Press, Chicago.
Vorontsova, M. A., and L. D. Liosner. I960 Asexual Propagation and Re¬ generation. Pergamon Press, Inc., New York,
31 32
Waddington, C. H. 1957 Principles of Embryology, George Allen and Unwin, Ltd., London, pp. 302—32U.
Weiss » P. 1939 Principles of Development. Henry Holt and Company, New York, pp. H5«-U78.
Articles
Protozoa Balumuth, W. 19h0 Regeneration in protozoa: a problem in morphogenesis. Quart. Rev. Biol., 15: 290-337*
Lillie, F. R. 1897 On the smallest parts of Stentor capable of re¬ generation, a contribution on the limits of divisibility of living matter. Jour. Morph., 12s 239-2U9.
Coelenterates (Hydra and Tubularia) Barth, L. G. 19I4.O The process of regeneration in hydroids. Biol. Rev., 15: U05-U20. Barth, L. B. 19UU The determination of the regenerating hydranth in Tubularia. Physiol. Zool., 17: 355-366. Berrill, N. J. 19U8 Temperature and size in the reorganization of Tubu¬ laria. Jour. Exper. Zool., 107: U55-U6U. Berrill, N. J. 1957 The indestructable hydra* Scien. Amer*, 197: 118-125* Brien, P. i960 The fresh-water hydra. Amer. Sci., lj.8: U61-U75*
Burnett, A. L. 1959 Histophysiology of growth in hydra. Jour. Exper. Zool., II4.O: 281-3U2.
Chalkey, H. W. 19U5 Quantitative relation between the number of organized centers and tissue volume in regeneration masses of minced body sections of Hydra, Jour. Nat. Can, Inst., 6: 191-195* Curtis, W. C. 19^0* The histologic basis of regeneration and reassociation in lower invertebrates. Amer. Nat,, 7U* U87-500. Goldfarb, A. J, 1909 The influence of the nervous system in regeneration. Jour. Exper. Zool., 7: 6U3-722. Ham, R. G. 1958 Chemically-induced loss of regenerative capacity in hydra. Fed. Proc., 17: 236. Ham, R. G., and R. E. Eakin 1956 Effects of lithium ion on regeneration of Hydra in a chemically defined environment. Jour. Exper. Zool., 133: 559-572. 33
Ham, R. G., and R. E. Eakin 1958 Time sequence of certain physiological events during regenerative in hydra. Jour. Bxper. Zool., 139s 33-51.
Lillie, F. R. 1900 The source of material of new parts and limits of size. Amer. Hat., 3U* 173-177.
Moore, J. A. 1939 The role of temperature in hydranth formation in Tubularia. Biol. Bull., 76: 10U-107.
Rafferty, K. A. 1935 The fates of segments from Tubularia primordia. Biol. Bull.. 108: 196-205.
Roudàbush, R. L. 1933 Phenomenon of regeneration in everted hydra. Biol. Bull., 6U: 253-258,
Steinberg, M. S, 1955 Cell movement, rate of regeneration, and the axial gradient in Tubularia. Biol. Bull., 108: 219-23U.
Tardent, P. 1959 Principles governing the process of regeneration in hydroids. In: Developing Cell Systems and Their Control (D. Rudnick, ed., 18th Growth Symposium), Ronald Press Co., New York, pp. 21—UU.
Tweedell, K. S. 1958 Inhibitors of regeneration in Tubularia. Biol. Bull., llU: 255-269.
Flat-worms (Planarla)
Bardeen, C. R. 1901 On the physiology of Planaria maculata. with especial reference to the phenomena of regeneration. Amer. Jour. Physiol., 5: 1-55.
Bmsted, H. V. 1955 Planarian regeneration. Biol. Rev., 30: 65-125.
Curtis, W. C., and L. M. Schulze 193U Studies on regeneration. I. Contrasting powers of regeneration in Planaria and Procotyla. Jour. Porphol., 55s U77-512.
Rulon, 0. 19U0 The environmental control of regeneration in Euplanaria. Amer. Nat., 7Us 501-512.
Santos, F. V. 1931 Studies on transplantation in planarians. Physiol. Zool., hi 111-161;.
Annelids (Earthworms)
Berrill, N. J. 1952 Regeneration and budding in worms. Biol. Rev., 27s U01-U38. 3U Gates, G. E. 19l*8 On segment formation in normal and regenerative growth of earthworms. Growth, 12: l6j^»l80.
Hyman, L. H. 19ljl Aspects of regeneration in Annelids. Biol. Symposia. 2: 21*1-256.
Liebmann, E. 191*3 New light on regeneration of Eisenia foetida. Jour. Morphol., 73: 583-610.
Moment, G. B. 191*6 A study of growth limitation in earthworms. Jour. Exper. Zool., 103: 1*87-506. Moment, G. B. 1950 A contribution to the anatomy of growth in earthworms. Jour. Morphol., 86: 59-72.
Moment, G, B. 1953 The relation of body level, temperature, and nutrition to regenerative growth. Physiol. Zool., 2o: 108-117.
Crustaceans (Crayfish)
Emmel, V. E. 1910 Differentiation of tissue in the regenerating crust¬ acean limb. Amer, Jour. Anat., 19: 109-156.
Needham, A. E. 1953 The central nervous system and regeneration in Crustacea. Jour. Exper. Biol., 30: 155.-159.
Vertebrates
Birnie, J. H. 1931* Regeneration of the taü-fins of Fundulus embryos, Biol. Bull., 66: 316-325.
Brynes, E. F. 1901* On the skeleton of regenerated anterior limbs in the frog. Biol. Bull., 7: 166-169.
Collins, H, H. 1932 Regeneration of hindlimbs in the vemilion-spotted newt, Triturus viridescens. Anat, Rec., 5U, Suppl., 58 (Abs.). Durbin, M. L. 1909 An analysis of the rate of regeneration throughout the regenerative process;. Jour. Exper. Zool., 7: 397-1*20.
Dent, J. N., and E. L, Hunt 191*8 The relationship of temperature to the rate of regeneration of epidermis in Triturus viridescens. Anat. Rec., 100: 651*.
Ellis, M. M. 1908 Some notes on the factors controlling the rate of re¬ generation in tadpoles of Rana clamitans-Daudin. Biol. Bull., ll*: 281-283. 35
Ellis, M, M. 1909 The relation of the amount of tail regenerated to the amount removed in tadpoles of Rana clamitans. Jour, Exper, Zool., 7ï 1*21-1*55.
Eycleshymer, A, C, 1907-08 The closing of wounds in the larval Necturus. Amer, Jour, Anat., 7* 317-325,
Goss, R. J,, and M. W, Stagg 1957 The regeneration of fins and fin rays in Fundulus heteroclitus. Jour, Exper, Zool., 136: 1*87-508.
Gidge, N, M,, and S. M. Rose 19kh The role of larval skin in promoting limb regeneration in adult Anura, Jour, Exper. Zool., 97? 71-93,
Morgan, T, H., and S. E, Davis 1902 The internal factors in the re¬ generation of the tail of the tadpole. Arch, f, Entw-mech., 15: 3lU- 318.
Needham, J, 1936 Biochemistry and causal morphology in amphibian re¬ generation, Science Progress, 31» Ul—5U,
Nabrit, S, M, 1938 Regeneration in the tail fins of embryo fishes (Opsanus and Fundulus), Jour, Exper. Zool., 79* 299-308.
Polezhayev, L. W. 191*6 The loss and restoration of regenerative capacity in the limbs of tailless Anphibia. Biol, Rev., 21s H4I—llt7,
Reed, M. A. 1903 The regeneration of a whole foot from the cut end of a leg containing only the tibia. Arch, f, Bitw-mech., 17: 150-15U.
Rose, S. M. 191*2 A method !>r inducing lirrib regeneration in adult Anura, Proc, Soc, Exper. Biol, and Med., 1*9* 1*08-1*10.
Rose, S. M. 19l*l* Methods of initiating limb regeneration in adult Anura. Jour, Exper, Zool., 95* 11*9-170,
Rose, S. M, 191*5 The effect of NaCl in stimulating regeneration of limbs of frogs. Jour, Morphol,, 77* 119-11*0.
Schotte, 0. E., and E. G, Butler 191*1* Phases in regeneration of urodele limb and their dependence upon the nervous system. Jour, Exper. Zool., 97s 95-121.
Schotte, 0. E., and M. Harland 19l*3 Amputation level and regeneration in limbs of late Rana clamitans tadpoles. Jour, Morphol,, 73* 329-363,
Thornton, G. S. 191*2 Studies on the origin of the regeneration blastema in Triturus viridescens. Jour. Exper. Zool., 89: 375-391.
Zelney, G, 1909 The effect of successive removal upon the rate of re¬ generation. Jour, Exper, Zool., 7* 1*77-512. 36
Zelney, G. 1909 The relation between degree of injury and rate of re¬ generation - additional observations and general discussion. Jour. Exper. Zool., 7î 913-961.
Zelney, C. 1909 Some experiments on the effect of age upon the rate of regeneration. Jour. Exper. Zool,, 7* 563-593»
Zelney, C. 1916 A comparison of the rates of regeneration from old and
from new tissue. Proc. Nat. Acad. Sci.f 2î I4.8I4.-U86.
Zelney, C. 1917 The effect of degree of injury, level of cut and time within the regenerative cycle upon the rate of regeneration. Proc. Nat. Acad. Sci., 3: 211-217. 37
Statements on the Objectives
Introduction
Perhaps more than any other science, biology is a staggeringly large collection of facts. This poses a great problem in an elementary course of biology because the mere accumulation of these facts is a large task
and leaves little time to see what they mean and how they fit together.
It is not unheard of for students to fail utterly to see the point...j
they may never see it, and the unhinged facts may soon wash away from their minds.^ One way out of such chaos of detail is to generalize, where
possible. Such principles (or generalizations) based on several facts
can therefore be stressed, rather than mere memorization of details with¬
out any understanding of the underlying idea.
The principles used in this unit were originally selected from 2 McKibben1s analysis and subsequently scrutinized by a group of biologists
in the Atlanta University Center. All agreed to the importance of these
principles and the significance of studies on regeneration as being directly
related to an understanding of these principles.
Principles of Biology Associated with Regeneration Phenomena
A. Principles: Regeneration is almost universal among living things. Prom the simple to the more complex animals, the abilities to regenerate lost parts and to reproduce asexually, fall off, gradually and independently, as the body becomes more specialized.
Growth and repair are fundamental activities for all pro¬ toplasm. Prom the lower to the higher forms of life,
Ï “ J. T. Bonner, The Idea of Biology (New York: Harper and Brothers, 1962), p. ix.
^M. J. McKibben, op. cit. 38
there is an increasing complexity of structure, and this is accompanied by a progressive increase in division of labor. In all organisms, the higher the organization the greater the degree of differentiation and division of labor and of the dependency of one part upon another.
Associated Phenomena: Apparent direct correlation between the ability of an organism to regenerate and the position it occupies on the evolutionary scale of animal organization. Quan¬ titative and Qualitative gradations in regenerative abilities between lowest and highest animal phyla.
B. Principles: Growth and development in organisms is essentially a cellular phenomenon, a direct result of mitotic cell division. Cells are organized into tissues, tissues into organs, and organs into systems, the better to carry on the functions of complex organisms.
All cells arise through the division of previous cells. Cell division is the essential mechanism of reproduction, of heredity, and to a large extent, of organic evolution.
Associated Phenomena: Source of the cellular materials involved in restoring lost cellular areas: Do they arise anew or are they produced by the transformation of cell types already present? Precise organization of the regenerated part.
C. Principles: The environment acts upon living things, and living things act upon their environment. Since the environment of living things changes continually, these creatures are continually engaged in a struggle with their environ¬ ment.
The range of temperature for life activities is very narrow as compared with the range of possible temperature. There is a minimum temperature below which, and a maximum temperature above which, no life processes: are carried on. The temperature range for life processes is.from many degrees below 0°C. to nearly the boiling point of water.
Associated Phenomena: The apparent control of regenerative rates by extrinsic forces. The re-initiating of regeneration, by external means, in animals that lose, after some time, their ability to regenerate certain parts of their body.
The re-awakening, by externally applied factors, of the ability of a seemingly well-determined and stable organism 39
to exhibit once again, capacities usually reserved for the embryo; that is, abilities of adult animals to restore lost and repair damaged parts.
D. Principles: Adult organisms that differ greatly from one another but which show fundamental similarities in embryological development, have originated from similar ancestors.
Animals resemble each other more and more closely the farther back we pursue them in embryological development.
Associated Phenomena: The strikingly different abilities for regeneration of lost limbs in such closely related animals (amphibians) as adult frogs and salamanders.
The strikingly similar abilities for regeneration of lost parts in the tadpoles (young stages) of frogs and salamanders.
E. Principles: The protoplasm of a cell carries on continuously all the general processes of any living body; the processes concerned in the growth and repair or upbuilding of protoplasm (anabolism) and the processes concerned with the breaking down of the protoplasm and elimination of wastes from the cell (catabolism). The sum of all these chemical and physical processes is metabolism.
Associated Phenomena: Apparent correlation between age of the organism and the capacity for regeneration, or the rate of regeneration.
Suggested Activities for Securing the Objectives
Introduction
The objectives previously stated are to be carried out by the
Question-Experiment method. The significance of experimentation in high school biology courses has gained significant support from the American
Institute of Biological Sciences. Glass has recently commented on this problem:
The biological sciences are currently advancing at so rapid Uo
a rate as to double the amount of significant knowledge in every ten to fifteen years. While this fact makes it imperative to revise our courses and methods of teaching at more and more frequent intervals, it also makes it increasingly impossible to •cover' in any course all that-is significant and that a general citizen might profitably know.
How can one truly understand the nature of science as in¬ vestigation and inquiry without some active participation in the experimental attack upon a new and unsolved problem? No matter how much you learn about the facts of science, you will never quite understand what makes science the force it is in human history, or the scientists the sorts of people they are, until you have shared with them such an experience. The laboratory and the fields are the scientists' workshops. Much reading and discussion are necessary in scientific work, but it is in the laboratory and field that hypotheses are tested. Properly to realize this aim, the student's experience must involve real, not make-believe, scientific investigation.
Questions on Regeneration
Question 1. Do "lower" (invertebrates) and "higher" (vertebrates) animal groups show the ability to regenerate?
Experiments on Invertebrates: Stentor - a protozoan Hydra - a coelenterate Planaria - a flatworm Earthworm - a segmented worm (annelid) Crayfish - an arthropod (crustacean) Starfish - an echinoderm
Experiments on Vertebrates: Goldfish - a bony fish Frog tadpole - an amphibian Mouse - a mammal
Question 2. Is there any correlation between the quantity (or extent) of regeneration and the simplicity or complexity of the organism?
Experiments on: Planaria - a flatworm Lumbricus - an earthworm (annelid) Frog tadpole - an amphibian
Question 3. When an animal is cut into two equal parts, does each re¬ generate attain the size of the original organism?
"^B. Glass, "Perspectives: A New High School Biology Program," American Scientist. XLIX (December, 1961), p. !?2J>. p. 529. la
Experiments on: Planaria - a flatworm Luribricus - an earthworm (annelid)
Question li. When part of an animal is removed, will the regenerated portion reach the size of the part originally removed?
Experiments on: Planaria - a flatworm Crayfish - an arthropod (crustacean) Lumbricus - an earthworm (annelid) v Frog tadpole - an amphibian
Question Is the ability of an organism to regenerate organized along an axial gradient (antero-porterior, posterior-anterior polarity)?
Experiments on: Planaria - a flatworm Tubularia - a coelenterate Earthworm - an annelid
Question 6. Can a ‘'fragment" give rise to a whole and thoroughly organized organism?
Experiments on: Planaria - a flatworm Tubularia (or Hydra) - a coelenterate Lumbricus - an earthworm (annelid)
Question 7. Does the type of cut made on the animal have any effect on the morphology of the regenerated part?
Experiments on: Planaria (a flatworm) longitudinal cut oblique cut transverse cut partial longitudinal oblique transverse
Question 8. Is the "material" for regeneration dispersed or localized within the animal?
Experiments on: Planaria a flatworm Tubularia (or Hydra) - a coelenterate
Question 9. Are extrinsic forces of significance in regeneration?
Experiments on the Effects of Food - Planaria Experiments on the Effect of Temper attire - planaria Experiments on the Effects of Crowding - Planaria
Question 10. Are intrinsic forces of significance in regeneration? k2
Experiments on the Effects of Age - Frog Tadpole Experiments on Previous History of Regeneration - Flanaria Experiments on Effects of Nerve Supply- Lumbricus
Question 11. May closely related organisms (evolutionary, i. e.) show different capacities for regeneration?
Experiments on Annelids: Earthworm (Lumbricus) and Leech Experiments on Amohibians: Adult Salamander and Adult Frog
Experiments for Answering Question 1
Stentor (a protozoan): Place s single protozian in a small amount of
water or culture medium on a depression slide on the stage of a bin¬
ocular microscope. With an £*ibryo knife or fine needle sharpened
to a thin blade, cut the specimen transversely into two parts (an
anterior and a posterior one) with a fairly quick motion. Add more
water or culture medium and place the slide in a moist chamber for
daily observation. More definite observations may be made if the cut
pieces are separated by using another depression slide (previously
marked) noting whether the piece is an anterior or a posterior part.
(See Figure 1).
Hydra (a coelenterate): Take three (3) Hydra and place each in a separate
container of culture fluid. Obtain a small flask of additional culture
and six more glass containers (small dishes). Label the dishes con¬
taining each Hydra A, B, C, Label the other dishes as follows: A-
base, A-hydranthj B-base, B-hydranth; C-base, C-hydranth. Pour into
each of the dishes some culture medium. Cut each hydra through a
region slightly below the hydranth (containing the tentacles), and
slightly above the base, thus leaving the stem portion. Place the base and hydranth pieces in their proper dishes and leave the stem portion ~har- h3 in the original dishes. Set the dishes aside and make daily ob¬ servations, Sketch any changes you may note at the cut surfaces of all of the pieces. Does each piece become a whole Hydra? How long does it take for complete regeneration to occur in each piece? (See Figure 2).
Flanaria (a flatworm) : Using a camel hair brush, place a single specimen in a small dish on the stage of the binocular microscope. Add a small amount of pond water, enough to cover the body surface. With a sharp scalpel, cut the specimen transversely into equal halves (i.e,, an anterior and a posterior part). Add more water. Label and cover the dishes (placing each piece in a separate dish and noting whether it is anterior or posterior). Record the progress of the cut pieces, (See Figure 3),
Lunibricus (earthworm; an annelid): Place a group of earthworms in a finger bowl filled with damp paper towel strips to remove the grit and dirt from the body surface of the animal, (The animals may be cleaned in groups,) Allow the animals to clean for thirty to sixty hours. Remove the animals from the cleaning bowl one at a time and place in a petri dish on the stage of a binocular microscope. Add a small amount of spring water, enough to cover the body surface. Two drops of chloretone (2 per cent solution) should be placed in the the medium to anaesthetize the animal. With a sharp razor blade, cut the specimen transversely into equal parts (anterior and a posterior part). Place each piece in an appropriately labeled bowl that contains dirt from the natural environment of the worm and cover the bowl with 33531 PïiC
• 2- ~ f-jyJr-<3 ï~i f 3 PUnar 'ta kh
damp, punctured paper towels. The toweling and dirt must be kept
moistened. Make daily observations on the pieces, (See Figure U).
Gambarus (crayfish: an arthropod crustacean): Take three (3) crayfish
and place them in separate jars containing a small amoung of water.
Label the jars A, B, C, Remove the antenna of the animal in jar A
by cutting through the point (X) shown in Figure 5, Sever a walking
leg (Point 0 of same Figure) of the animals in jars B and C, Set
jars aside and record the changes which take place in the animals
whose antennae and walking legs (portions) have been removed,
Asterias (starfish; an echinoderm): Sever two (2) starfishes in the
manner described in Figure 6, Place the pieces in appropriately
labeled containers and set aside for subsequent inspection. Record
all changes and make sketches of any external signs of restoration.
Goldfish (a bony fish): Take three (3) goldfish and place each of them in
a separate container of water. Remove and discard the t ail fin of
each animal (see Figure 7), Set the containers aside and record all
subsequent changes.
Frog tadpole (an amphibian): Place a single tadpole in a petri dish con¬
taining no pond water. With a sharp razor blade cut off and discard
about 2ram from the posterior most end of the tail. Place each tadpole
in a separate battery jar containing pond water and record all sub¬
sequent developments, (See Figure 8.)
Mouse (a mammal): Place a mouse into a glass jar. Close the jar opening
with a top to which a wad of cotton has been attached. Prior to
/Ai/oiwa f-ïf 7- QroUfiïU
fc f- ~ TjdjOo/a. closing, the cotton should be dipped in ether. In this way an--
aeâthètizethe mouse but do not kill it by allowing too long an ex¬
posure to the ether. When the mouse has been overcome by the ether
fumes, remove it from the jar and shave a region of the dorsal side
of the body. With a sterile, well sharpened scalpel, inflict a
skin-deep wound within the shaven area. Return the animal to a
separate container. Record superficial changes in the wounded area.
Experiments for Answering Question 2
Flanaria (a flatworm): Using a camel hair brush, place a single specimen
in a small stender dish on the stage of a binocular microscope.
Add a small amount of pond water, enough to cover the body surface.
With a sharp razor blade cut the tip of the posterior end (approxi¬
mately one-fourth of the total body size). Add more culture medium
(pond water). Label and cover the dishes. In all cases, place the
posterior piece in one dish of water and the anterior three-fourths
in another. Set aside and record all observations,
Lumbricus (earthwormj an annelid): Place a group of earthwoms into a
finger bowl filled with damp paper towel strips to remove the grit
and dirt from the body surface of the animals. Allow the animals to
clean for thirty to sixty hours. Remove the animals from the cleaning
bowl one at a time and place in a petri dish on the stage of a
binocular microscope. Add a small amount of pond water, enough to
cover the body surface. Two drops of chloretone (2 per cent
solution) should be placed in the medium to anaesthetize the animal.
With a sharp razor blade, cut the tip of the posterior end (approximately U6
one-fourth of the total body length). Add more culture medium
(pond water). Each piece should be placed in appropriately labeled
finger bowls containing dirt from the natural environment and the
bowls kept covered with damp, punctured paper towels, Make periodic
observations on the progress of each cut as it attempts to restore
itself.
Frog Tadpole (an amphibian): Place single tadpole in a petri dish con¬
taining no pond water. With a sharp razor blade cut through the tip
of the posterior end (appriximately one-fourth of the total body
length). Discard the severed tip; place the amputated specimen into
a battery jar containing pond water, and record all subsequent de¬
velopment.
Experiments for Answering Question 3
Planaria (a flatworm): Using a camel hair brush, place a single specimen
in a small dish on the stage of a binocular microscope. Add a small
amount of pond water, enough to cover the body surface. With a
sharp razor blade, cut the animal transversely into equal parts (thus
an anterior half and a posterior half). Add more spring water.
Place each half in a separate container, appropriately labeled. Prior
to cutting, measure the length of the planarian. Record all sub¬
sequent transformations of the cut pieces,
Lumbricus (earthworms an annelid): Place a group of earthworms into a
finger bowl filled with damp paper towel strips to remove the grit
and dirt from the body surface of the animals. Remove a single
specimen from the cleaning bowl and place in a petri dish on the 1*7
stage of a binocular microscope. Add a small amount of spring water,
enough to cover the body surface. Two drops of a 2 per cent
chloretone solution should be placed in the medium to anaesthetize
the animal. With a sharp razor blade, cut the animal transversely
into two equal parts. Each half is then placed into a bowl, ap¬
propriately labeled, that contains dirt from the natural environ¬
ment and covered with danqp, punctured paper towles. Keep an accurate
record of the periodic observations made on the progress of each cut
half.
Experiments for Answering Question It
Planarian (a flatworm): Using a camel hair brush, place a single specimen
into a stender dish on the stage of a binocular microscope. Add a
small amount of pond water, enough to cover the body surface. With
a sharp razor blade, cut one side of the anterior half of the
organism, as shown below (a). Another animal may be cut, as in (b),
while another as in (c). Place all pieces in appropriately labeled
dishes containing some pond water, ^ake daily observations and
record all changes.
Lubricus (earthwormj an annelid): Place a group of earthworms into a
finger bowl filled with damp paper towel strips to remove the grit and dirt from the body surface of the animals. Remove the animals
from the cleaning bowl one at a time and place in a petri dish. Add
a small amount of pond water, enough to cover the body surface. Place
two drops of 2 per cent chloretone solution into the medium to an¬
aesthetize the animal. With a sharp razor blade, cut off five
segments from the anterior end of an animal marked A, and ten segments
from the posterior end of an animal marked B. Prior to cutting, make
a record of the number of segments anterior to the clitellum and the
number posterior. In this way, should regeneration occur you will
know if the correct number of segments has been restored. Place all
pieces in correctly labeled bowls containing dirt from the natural
environment and covered with damp, punctured paper towels. Record
all periodic inspections.
Cambarus (crayfishj an arthropod crustacean): Place a crayfish into a
finger bowl containing pond water. With a sharp razor blade, cut
off and discard all the right or left walking legs at various levels.
Place the amputated specimens into battery jars (or jointly into an
aquarium), with proper labeling on each jar. Observe the animals
every two or three days and record all observations.
Frog Tadpole (an amphibian): Place a single tadpole into a petri dish
containing no pond water. With a sharp razor blade, cut off 3-5mm
of the posterior part of the tail. Put the amputated tadpole into
battery jars containing pond water, set jars aside, and make daily
observations. Experiments for Answering Question $
Tubularia or Hydra (coelenterate): Cut a single specimen into several
parts (as indicated below). Give each cut region a label and place
each niece into a separate container of sea water (for Tubularia)
or de-ionized water plus Calcium (for Hydra), talcing care to label
each container appropriately. Set the containers asi.de and make
hourly observations. Does each piece form a complete organism,
having a stem, base and a hydranth?
Planaria (a flatworm): Cut a single planarian into several parts (as
diagrammed below). Give each region a label and place all pieces
into separate containers of pond water. Each container should be ap¬
propriately labeled. Set the vessels aside and make periodic ob¬
servations on the progress of each piece. Do extreme tail pieces
form a head? Do extreme head, pieces form a tail? Are the middle
regions more capable of forming both head and tail?
Lumbricus (earthworm; an annelid); Anaesthetize an earthworm with 2 per
cent chloretone and cut into several pieces according to the diagram 50
below. Place each piece into an appropriately labeled dish containing
some dirt from the natural environment of the worm. Cover the dishes
with damp, punctured paper towels. Record all subsequent observations.
Are the posterior-most segments capable of forming heads? Are the
anterior-most segments capable of forming tails? As one progresses
from the anterior to the posterior segments, in which regions will
both head and tail- formation occur most frequently?
Hydra or Tubularia (coelenterate): Cut out a fragment of the stem of one
of these hydroids, as indicated below. Discard the anterior part
containing the hydranth and the posterior part bearing the base.
Place the fragment in a separate dish of culture fluid (sea water or
de-ionized water with calcium added) and make frequent observations.
Sara.
Planaria (a flatworm): Cut out a fragment of the worm, as indicated below.
Discard the other portions. Place the fragment in a dish of pond
water, set aside and make daily observations. ^ 51
Lumbricus (earthworm; an annelid): Cut out a piece of the worm from the
posterior-most end of the clitellum to twenty-five segments back.
Discard all other pieces. Return the fragment to a dish of moistened
dirt (kept moist with damp paper towels). Record all subsequent
observations.
Sa-r-c,
Experiments for Answering Question 7
Planaria (a flatworm): At least seven (7) planarias are needed in this
experiment. ’The worms should be labeled 1-7. The manner of
cutting for each worm is described in the diagram beloxj: 52
Worm Number 1 - Arrow A - B
Worm Number 2 - Arrow C - D
Worm Number 3 - Arrow E - F
Worm Number k - Arrow A - 0 (and or B - 0)
Worn Number 5 - Arrow C - 0 (and or D - 0)
Worm Number 6 - Arrow E - 0 or F - 0
Worm Number 7 - Zig-Zag line indicated by Arrow X - XX
Keep both pieces (where necessary) in the same container of pond
water. Make daily observations and record all changes.
Experiments for Answering Question 8
ELanaria (a flatworra): Gut a single planarian into as many parts as are
shown in the following diagram. Label each piece and place it in an
appropriately labeled dish containing pond water. Set aside and
make daily observations. What is the fate of each piece? Does each
piece become a whole planarian? 53
Experiments for Answering Question 9
Effects of Food: Use Planaria (a flatworm) - Starve five planarias for
varying periods of time ( two days, five days, seven days, etc.).
Gut the starved animals transversely into an anterior and a posterior
half. Place each cut piece into a separate container of pond water
and label each appropriately. Make daily observations. Compare the
time required for total generation to occur with the animals re¬
generating according to Experiments for Questions three on planaria.
Take three planarias that have been allowed to grow in a medium
containing an excess amount of food (such that the animals are never
without food) for a period of four to seven days. Cut the overly
fed animals into equal halves with a transverse cut. Place each
piece into a container of pond water, with each container being
appropriately labeled. Make daily observations. Compare the time
required for total regeneration to occur with that in Experiments
for Questions three and nine on planaria.
Effects of Temperature: Use Planaria (a flatworm) - Place pieces of
transversely cut planarias into appropriately labeled containers of
pond water and allow subsequent development to occur at the following
temperatures: 3 “ 5°C. 10 - 15°C. 2h - 26°C. 27 - 28°C. 29 - 30°C. 32 - 3U°C.
Make daily observations and compare the time required for total
regeneration to occur with the time required for the planarias in Question three to regenerate,
Effects of Crowding (Population Density): Use Planaria (a flatworm) -
Repeat the Experiments on Planaria in Question five. Instead of
placing each piece into a separete container, place all pieces into
a single container of the same size, Hake daily observations and
compare time required for total regeneration to occur with the value
obtained for the regenerating planaria in Question five.
Experiments for Answering Question 10
Effect of Age: Use Frog Tadpole (an amphibian) - Repeat the Experiment
on the Frog Tadpole in Question four, using tadpoles of different
ages (based on total body lengthj the older ones being longer and
larger). Place each tadpole in a separate container, taking care
to label each vessel appropriately. Make daily observations and
compare the time required for tail regeneration to occur with values
obtained from Question four.
Effect of Previous History of Regeneration: Use Planaria (a flatworm) -
Repeat Experiments on planaria in Question three several times,
always using one of the regenerated organisms in subsequent cuttings.
Make a careful study on the time required for total regeneration to
occur in a First Regenerate, Second Regenerate, Third Regenerate,
Fourth Regenerate, and a Fifth Regenerate, Do Fifth Generation Re¬
generates become restituted at the same rate as First Generation
Regenerates?
Effect of Nerve Supply: Use Lumbricus (earthworm; an annelid) - With 55
a sharp razor blade, make a partial transverse cut in an earthworm
as indicated by Arrow A (Figure U). As seen in the Figure, this cut
will not reach the ventral nerve cord. In another earthworm, make
a partial transverse cut as indicated by Arrow B in Figure [>, This
cut will penetrate the ventral nerve cord. Place each animal in an
appropriately labeled container of moistened dirt from the environ¬
ment of the animal, hake daily observations and record changes
occurring in both.
Experiments for Answering Question 11
Annelids (an earthworm and a Leech): Make transverse sections (as indi¬
cated by the arrows in the diagram belox*) through an earthworm and a
leech. Place each piece in an appropriately labeled container. Set
aside and make daily observations. Record all changes. Do the earth¬
worm and leech fragments regenerate, giving complete worms?
Amphibians (a salamander and a frog): Cut off the forelimb of an adult
salamander (as shown in Figure A below) and an adult frog (as shown
in Figure B), Place each amputated animal in a separate jar, set
aside and make periodic observations. Record all changes. Do both of
these animals show similar abilities to restore portions of amputated
limbs? 56
Evaluation of the Experiences from the Activities of the Unit
Ability to Accurately Observe and Record Data»—Make a constant check on each student to determine the sharpness of his ability to make ob¬ servations. Watch for growth in the student’s ability to make careful observations* ones which do not often "meet the eye” at first glance.
Have each student turn in periodically a written record of his observations and the analysis of his data. Stress throughout the activity the necessity for scientific accuracy in recording data.
Operation Skills Developed.—Always be cognizant of the skills of each student in making proper operations* The student is to be made aware of the importance of handling live materials and the care one must take in carrying out his dissecting.
Ability to Propose few Experiments for Testing Validity of Other
Principles not Included in This Unit.—Have the student propose new ex¬ periments to test the validity of other biological principles not covered
in the unit.
Examination.—Prepare an examination to test how well the student has
grasped, not only the broader picture of the experiments and their results, but details as well.
Appendix for the Unit
Animals Required
The following animals represent suitable experimental material for 1 studies on regeneration:
Ï " Organisms 1-8 are invertebrates, while 9-12 are vertebrates. Si
1* Stentor - a ciliated protozoan
2. Hydra - a fresh-water coelenterate
3. Tubularia - a marine coelenterate
lu Planaria - a free-living flatworm
5* Earthworm - a free-living annelid (segmented) worm
6, Leech - an ecto-parasitic annelid (segmented) worm
7. Crayfish - a crustacean (arthropod)
8. Starfish - an echinoderm
9, Goldfish - a bony fish
10. Salamander - a tailed amphibian
11. Frog (adult and tadpole) - a tailless amphibian (adult, that is)
12. Mouse • a mammal (rodent)
Methods for Culturing and Caring for Animals in the Laboratory
The accompanying leaflets, published by the General Biological Supply
House,^ will serve as appropriate guides for laboratory care of the animals required for the experiments suggested in this Unit, There is a leaflet for every animal listed above, except Tubularia and Starfish and Goldfish.
Tubularia and Starfish live in a marine environment} hence, the leaflet entitled "Notes on Marine Aquaria" should be used for these organisms* The leaflet entitled "Starting and Maintaining a Fresh-Water Aquarium" will suffice for the Goldfish* General operation techniques are cited in the leaflet on "Laboratory Dissection." In addition, since some of the animals thrive on algae, there is a special leaflet on "Growing Fresh-
Water Algae in the Laboratory."
Ï “ General Biological Supply House, 8200 South Hoyne Avenue, Chicago 20, Illinois. î>8
Equipment and Supplies Needed
All of the equipment and supplies called for in executing the experi¬ ments may be obtained from the General Biological Supply House* 8200 South
Hoyne Avenue, Chicago 20, Illinois. The leaflet "Basic Laboratory Equip¬ ment for High School Biology Course" contains the pertinent information for ordering these and other supplies. Another leaflet, “Preserving
Zoological Specimens: Narcotization, Fixation and Preservation", provides adequate information about narcotizing (or anaesthetizing) the animals prior to operation. Furthermore, should anyone desire to preserve a specimen used in his experiments, appropriate suggestions are provided in the textual materials included in this unit.
Non-Specific Items
In order to provide further assistance to the teacher who may desire to do additional work not cited in the Unit, several other leaflets are included: "Special Projects for Biology Students," "Demonstration and
Display Materials," "Embryology in the High School Biology Course," "A
Selected List of Books for the Biology Library." TURTOX SERVICE LEAFLET No. 4
THE CARE OF PROTOZOAN CULTURES IN THE LABORATORY
Care of Cultures our laboratory in excellent condition but Sometimes encounter delays or are ex¬ Protozoa are almost infinite in form, in habit and in their distribution, and a posed to temperature extremes which wide range of kinds can be found in any kill the specimens. Cultures received hay infusion or culture of decaying pond in poor condition should be reported at weeds. Simply because such “Mixed Cul¬ once and at the same time a delivery tures” contain a great many forms which date should be given for the replacement. are not easily recognized is no argument If no delivery date is indicated the re¬ for ignoring them in the school labora¬ placement will be made promptly. tory. Their variety and the changes in After the cultures have been received protozoan population which occur from and opened allow them to remain un¬ day to day, the cycles through which a disturbed for an hour or more. In search¬ mixed culture passes and the great num¬ ing for the protozoan in the culture jar ber of different forms present, are all the characteristics of the species should be extremely interesting to one who wishes kept in mind and much fruitless searching to know something of the science which with pipette can be eliminated. Ameba we call Biology. settle to the bottom or on the sides of Field collecting in any pond or stream tlie container. Paramecium, Euglena, will produce an abundance of protozoa Didinium, Blepharisma, Euplotes and from which pure cultures of some forms Colpidium are found swimming freely may be obtained. This takes time, how¬ throughout the entire culture. They can ever, for after inoculation a pure culture usually be concentrated by wrapping the usually requires several weeks to develop bottle with dark paper leaving only the to a point at which it is best for labora¬ surface of the water exposed to the light. tory study; and, furthermore, a “pure” Stentor and Vorticella attach themselves culture seldom remains pure for any to food, sides and bottom of the bottle. long period of time. To check the protozoan culture or study Since laboratory work is given at the the forms in class place a drop of the beginning of the fall term, and it takes culture water on a clean microscope slide time to rear protozoan cultures, the and cover with a clean coverglass. Ex¬ teacher finds himself faced with the prob¬ amine under the low power objective and lem of supplying his students with living be sure the light is well stopped down. material at the very beginning of the Cultures of protozoa used in class work school year. It is usually advisable, there¬ usually contain more specimens than fore, to purchase pure cultures from Tur- needed and the remaining material can tox and keep these going as long as pos¬ he used in propagating cultures in the sible after they have served their purpose laboratory. The same procedure can be in the course. used in starting pure cultures by isolat¬ Turtox cultures are shipped in tightly ing species found in a mixed protozoa capped two-ounce jars. As soon as they collection. are received at the school the screw- In culturing protozoa the important caps should be removed and the bottles factors to consider are temperature, light, kept at a temperate of not above 70° F kind and quantity of food, type of culture and out of direct sunlight. The cultures jar, and freedom from contamination. can be left in the shipping jars until These factors vary widely of course and they are used in the laboratory. Anieba, will be treated separately for each specific Paramecium, Euglena, Stentor and form. others will keep in good condition for days without any attention, but mixed Ameba protozoan cultures should be studied If the teacher wishes to culture Amebae immediately since the larger forms eat from a local source, some decaying water the smaller and the variety is quickly plants (water lily leaves are particularly reduced. good) and pond water should be ob¬ Material which has been purchased tained. A little of the plant material and should be examined immediately upon about 50 cc of pond water may be placed unpacking to be certain the protozoans in a finger bowl dish and observed daily. have not died in transit. Shipments leave To save time it is usually best to start
TURTOX Service Department Copyright, 1959. by GENERAL BIOLOGICAL SUPPLY HOUSE (INCORPORATED) 8200 South Hoyne Avenue Chicago 20, Illinois THE SIGN OF THE TURTOX PLEDGES ABSOLUTE SATISFACTION Printed in U.S.A. GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
several cultures, using pond weeds and found in nature only five times since water from several localities. If Ameba then; in 1900 by H. V. Wilson in North are present, they should appear in quan¬ Carolina; in 1902 by E. Penard in Swit¬ tities in about two weeks, being most zerland; in 1916 by W. A. Kepner in plentiful on the bottom of the culture Virginia; and by A. A. Schaeffer in Ten¬ dish. In looking for them, pipette a few nessee and by A. A. Schaeffer in 1936 in drops of the bottom sediment onto a a marsh in New Jersey. Since rediscover¬ slide and examine under the microscope. ing this ameba in 1936, Dr. Schaeffer has Ameba appears to do best with a mod¬ succeeded in culturing it and, during the erate temperature, diffused light and very 1937-38 school year, his pure cultures little food. At 50° F. they are inactive were made available to hundreds of teach¬ and sluggish, at 75° F. they develop rap¬ ers—General Biological Supply House idly and at a temperature of 90° or acting as Dr. Schaeffer’s exclusive dis¬ higher there is little chance of their sur¬ tributor. vival. Chaos chaos is of a very large size, fre¬ Direct sunlight is harmful to Ameba quently attaining a length of from 4 mm and an over abundance of light encour¬ to 9 mm. when in locomotion. It is easily ages the development of the other more seen with the unaided eye. However, this vigorous protozoa, such as Euglena and large size causes the amebas to break up Paramecium, which result in rapid con¬ in shipment and there seems to be no way tamination of the culture. As a general of preventing this fragmentation which rule, windows located on the side of the occasionally occurs. The result is not a room opposite that side on which the dead culture, but a living culture which cultures are placed will provide suf¬ contains small instead of large amebas. ficient light for Ameba. Ordinary stacking finger bowls make Caring for the Culture. Unscrew the excellent culture dishes. Several dishes culture jar immediately and allow it to may be stacked one upon the other and stand undisturbed for an hour in a cool the top dish may be covered with a piece place (60° to 70° F.), not in strong light. of glass. Wheat kernels and pieces of Then examine the culture, looking for the timothy hay about 2 inches in length, amebas under a dissecting microscope or which have been boiled for five minutes, under the lowest power of the compound supply the source of food. microscope. If large specimens are pres¬ Each bowl is filled with 75cc. distilled ent, they will be seen without difficulty. If water to which 1 grain of wheat (cut in the amebas appear to be of small size, two) and one 2 inch piece of hay (cut allow the culture to remain in the un¬ into )4" lengths). Using a clean pipette, capped jar for three or four days; in inoculate immediately with amoeba from most instances they will increase in size a pure culture. After 2 weeks discard greatly during the period. one half of the fluid and add enough Sub-culturing. Chaos chaos is a heavy distilled water to bring the volume to feeder and for rapid growth it must have lOOcc. Add the same amount of food as abundant food such as Paramecium or at the first if the culture is “clean”— other ciliates. New cultures of Chaos with not too many paramecium and not chaos can be started by the method com¬ too much bacterial growth. Reduce the monly used for culturing Paramecium. amount of the second feeding if the When the new culture is rich in Parame¬ culture shows these signs of over-rich¬ cium inoculate with the Chaos chaos. In ness. An even simpler medium consists of just three grains of boiled wheat per three to six weeks the culture will show good growth. culture dish treated in the same way as indicated above. Study Suggestions. Examine this ameba These cultures are examined every 2 in a shallow culture dish, with the naked weeks to observe the food supply and eye and with a low power (5X) magnifier. the abundance of the Amebae. The water Then observe under the low power of the mold which invariably develops on the compound microscope, and study a por¬ wheat and hay is not detrimental to the tion of one specimen under high power. culture. After six weeks the Ameba Note the many nuclei. Using a microm¬ will have divided and increased to make eter eyepiece (or any other convenient a heavy culture and may now be used for method) measure the length of a speci¬ class study. If the cultures have been men “at rest” and the length of one in inoculated with one of the larger strains, locomotion. Study its method of engulfing the Ameba can be seen plainly with the (eating) a Paramecium. naked eye as tiny milky-white specks on the bottom and side of the dish. Paramecium Chaos chaos The culturing of Paramecium is not Chaos chaos, the largest ameba known to so difficult as culturing Ameba. They science, was first seen by Roesel von require little attention and develop very Rosenhof in Germany in 1755. It has been rapidly. Ordinary room temperature is
(4-2) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
satisfactory for good continuous growth acid for a short time, rinsed well with of the culture, but a temperature of 80° distilled water and then allowed to dry. to 85° will bring the culture to its maxi¬ Obtain clean dry timothy hay and cut mum growth in a shorter time. The best the spikes as well as the stems and leaves culture dishes for Paramecium are finger into segments approximately one inch in bowls, both the 4% and 7% inch diam¬ length. Place 1 to iy2 grams of the cut eter, because of the greater surface area hay into 250 cc. Erlenmeyer flask, add and less depth of water. However, any a knife-tip of precipitated CaCOa, fill to type of glass container will do for this the neck with distilled water, then cover protozoan. Timothy, rice and wheat the mouth of the flask with an inverted boiled about five minutes are good food snugly fitting beaker. Flasks so prepared for the culture. The culture will grow in are heated and the contents boiled for 15 direct and indirect light; however, the minutes. The flasks or cooked infusion latter is best because direct sunlight en¬ are now allowed to stand (ripen) for ap¬ courages other protozoa and results in proximately 24 hours, then the medium is contamination. ready for use. This medium is an ex¬ The dishes for culturing should be cellent one for the culturing of Parame¬ thoroughly cleaned. If the 4y2 inch di¬ cium caudatum, P. multimicronucleatum ameter finger bowls are used fill the and P. trichium. bowl two-thirds full with distilled water Although Paramecium bursaria and and add 4 kernels of boiled wheat or P. aurelia will grow in dilute hay infusion rice and 12 to 15 pieces of cooked better results, especially in demonstrat¬ timothy 15 to 20 mm. long. If larger ing the mating reaction, can be obtained finger bowls, or other containers are by using the lettuce infusion as described used, increase the amount of food in by Drs. Jennings and Sonneborn. This proportion to the amount of water. may be prepared by obtaining a clean The media are ready for immediate head of lettuce and placing the individual inoculation with paramecium from a leaves in an oven. When the leaves be¬ concentrated culture found locally or come brown and brittle they are crushed purchased from a reliable source. The to a powder with a mortar and pestle. older method of allowing media to age One and one-half grams of the desiccated prior to inoculation is no longer recom¬ lettuce powder is added to one liter of mended since there is often an observ¬ distilled water and boiled for five min¬ able tendency for the food organism utes then filtered into 250 cc. flasks while (bacteria) to outgrow and crowd out hot, stoppered with cotton and if pos¬ the cultured organism. The film of bac¬ sible, autoclaved. While the flasks are still teria which sometimes forms on the warm (but not hot) small square pieces surface of the medium should be broken of “parafilm” are used to cover and seal up whenever it is encountered. Its con¬ the plugged mouths of the flasks. This tinuance is detrimental to paramecium. is the stock fluid and flasks so prepared Stack finger bowls to any desired may be stored for weeks in the refrigera¬ height, cover the top one with a glass tor. Equal parts of distilled water and plate and place where they will receive the stock fluid are mixed and when the plenty of indirect light. mixture reaches room temperature (about Three or four days after inoculation 21° C.) the fluid is ready for use. the Paramecium will be seen concen¬ Flasks of pure-line mass cultures of trated near the surface of the water Paramecium may be kept on shelves at and around the food. In ten days or room temperature for months and still two weeks the concentration will be at contain many individuals. To maintain its maximum. When the Paramecium the pure-line culture it is only necessary begin to die down a little boiled timothy, to inoculate new flasks of infusion. If wheat or rice should be added to main¬ larger numbers of the ciliates are needed, tain the culture. Such cultures can be use large battery jars and, of course, kept in pure condition from four to six proportionately larger amounts of hay weeks. and distilled water. Demonstrating the Mating Reaction and Conjugation Pure-line Cultures of Paramecium for Two pure-line cultures of Paramecium General Study and Demonstration of bursaria are used for this demonstration. Conjugation These two cultures are mixed together, Practically all species of Paramecium preferably at or about noon on a sunny grow well in hay infusion. Because of the day. Immediately a few are seen to stick ease with which this culture medium can together, then the clump becomes larger be made, full directions will be given. and larger until clumps of many indi¬ All glassware that is to be used must be viduals are formed. These large clumps scrubbed clean and sterilization should be then become smaller until, after about done by autoclaving. If an autoclave is six hours, only small masses of individu¬ not available, glassware after it is als will be observed. Finally individual scrubbed may be placed in 10% nitric pairs of conjugants are present and re-
(4-3) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS main joined for from 24 to 48 hours. culture medium is prepared similar to that It is well to allow two days for this suggested for culturing Paramecium. demonstration, for the individual con¬ These forms may be easily isolated from jugating pairs will sometimes be best on the pond collection and used to inoculate the second day. The demonstrations the prepared culture medium. They de¬ should be made in watch glasses or other velop rapidly and will have increaseu shallow dishes so that the entire process considerably a week after inoculation can be studied under a low power micro¬ when they will be found in large numbers scope. IMPORTANT ! Use this material in the surface scum and upper half of the as soon as possible ! Results cannot be culture. Sub-cultures should be made guaranteed unless the conjugation cul¬ every three weeks. tures are used within 24 hours after Didinium is an interesting protozoan delivery. because of its carnivorous habits. It lives Euglena mainly on Paramecium. Consequently Euglena is the easiest protozoan to cul¬ paramecium are necessary in the culture ture because it requires plenty of food, of Didinium. These paramecium cultures light and little attention. Battery jars are developed as previously suggested or other tall jars are excellent for cultur¬ and when the paramecium are abundant ing and a window sill is an ideal place the Didinium can be added. The culture to keep cultures since direct sunlight is will last as long as there is a sufficient food beneficial. The temperature can range supply. About every ten days more from normal to 90° with little damage paramecium should be added or new to culture. Boiled wheat, rice or timothy paramecium cultures should be inocu¬ can be used for food. lated. A clean 6x8 inch battery jar should be The culture of the slow moving peach- filled with distilled water and a large colored protozoan, Blepharisma, can best handful of timothy or 100 kernels of be carried on in shallow culture dishes in wheat or rice added. Place culture on media similar to that used for Ameba. window sill and after a week add a few A little more boiled wheat is necessary drops of concentrated Euglena from a because Blepharisma grows better in rich pure culture which can be collected very cultures. Inoculation can take place two easily in stagnant pools. In two weeks weeks after culture has been started. the entire culture will begin to have a Blepharisma develop quickly in cultures greenish appearance and the surface of and require little attention from four to the water will be covered with a scum six weeks. They are usually found containing large numbers of Euglena. swimming slowly about near the bottom The culture will last for four weeks of the dish. without any attention. If more food is Arcella is common in most old pond- added every two weeks the culture will water cultures and will often be found last three or four months. on the bottom of the dishes containing Stentor old hay infusions from which the para- The culture of Stentor can be carried moecia have died out. Arcella is slow on in any type of disli such as finger moving and easily studied, but the light' bowls or battery jars. Because Stentor should be carefully regulated when ex¬ thrive in rich protozoan cultures contain¬ amining them with the compound micro¬ ing such forms as Euplotes, Colpidium scope. and Chilomonas, it is best to prepare “Both wheat and hay infusions are media with the idea of mixed protozoa good, but a mixture is best. To 100 cc. cultures in mind. This can be done fol¬ of pond water, add 2 grains of wheat lowing tlie same procedure as that given and y2 gram of hay. Inoculate with for Paramecium. When the cultûre is Chilomonas if available, although Arcella ready for inoculation, instead of using will grow by feeding merely on the de¬ Paramecium, use mixed protozoa. When composing infusion. After two or three the culture is well established add the days, add Arcella which may be found Stentor, keep the culture in diffused on the bottom of many old cultures or light and in a temperature of 65“ to in the bottom ooze of any shallow pond. 75°. It will be necessary from time to Isolate them from the ooze and other time to sub-culture to new media. Protozoa with a fine pipette and place The Culture of Other Protozoa them in the culture in a shallow dish.” Sometimes Studied —John P. Turner, University of Min¬ Before concluding the discussions on nesota. Quoted from “Culture Methods culturing protozoa, we believe a few ad¬ for Invertebrate Animals” by permission ditional remarks about the culturing of of Comstock Publishing Company, Inc. less familiar forms might prove worth¬ Culture Concentrates. while. Turtox concentrated media for the cul¬ Colpidium, Euplotes and Halteria are ture of protozoans, algae and such forms commonly found in pond collections and as Volvox and Chlamydomonas, is now can be grown in pure culture as easily as available. Write for price list and for the the other types already discussed. The Special Turtox Bulletin No. 61V170.
CM) TURTOX SERVICE LEAFLET No. 39
THE FRESH WATER HYDRAS The hydras are of interest to almost microscope red crustacean upon which every teacher of Biology for several rea¬ they had been feeding. sons. Hydras are studied in practically As pictures of hydras and discussions every beginning Biology and Zoology of their structures are given in nearly course and are also of unusual interest every Zoology text, these will not be in¬ because they are the only common fresh¬ cluded here. water representatives of the group of Coelenterata to which the marine jelly¬ Occurrence. fishes and corals belong. Many teachers, Hydras are found in nearly all perma¬ however, show their students only pre¬ nent bodies of fresh water, but are much served specimens in alcohol or hydras more abundant in some places than in mounted on slides, other localities where overlooking the fas¬ First Things to Do Upon the conditions are cinating studies Receipt of Hydra seemingly just as which are possible if (1) Before opening jar, shake it favorable for their a few living hydras gently to detach Hydra which growth. In general are available in the may be attached to the glass they are likely to be laboratory. or cap above normal surface found in lakes, slow- of water. flowing rivers and Form and (2) Open jar and allow it to reservoirs, rather Appearance. stand quietly for a few than in springs Hydras are not minutes. and swiftly-flowing large but after one (3) If Hydra are to be used streams. has become familiar soon, leave them in the jar, with their appear¬ placing it in a cool location. Where to Collect. ance and knows (4) Do not change the water in It will be rather where to look for the jar unless it appears difficult for the be¬ them they can be cloudy. If new water must ginner to locate Hy¬ seen and studied be added, use only clear dra in the field as fairly well even aquarium or pond water. the specimens con¬ without the use of (5) To keep Hydra permanently, tract when the wa¬ a lens. A hydra has release them in an aquarium ter weeds to which been described as which contains growing plants. they are attached are resembling a “short removed from the piece of string with one end frayed out,” water. Look for this form in permanent and this description is a good one if the pools or small lakes or even flowing student is made to realize that a very streams or rivers. In the Chicago area small bit of string is meant. The frayed one species is found upon the rocks along ends are the tentacles which radiate out the shores of Lake Michigan where the from the mouth region of the animal. waves continually pound the shore. When When fully expanded a hydra may be an collecting look for Hydra on the sub¬ inch or more in length and when merged water weeds. Place a few of the contracted the entire animal looks like weeds in a quart jar of clean pond a tiny drop of flesh-colored jelly. water and examine by holding it up to There are several species of Hydra, the light. The specimens will usually re¬ all of which are similar in general ap¬ main contracted for some time and will pearance, although they may vary greatly appear as tiny lumps of jelly with the as to color. The commoner (and larger) shortened tentacles projecting outward species are grayish, yellowish or brownish from the free end of this lump. Do not in color, although one hydra (Hydra give up the quest quickly, but make a viridissima) is a vivid green. When fully thorough search. At certain seasons of expanded the grayish and brownish kinds the year the Hydra may be very abund¬ are nearly transparent. Food sometimes ant, while at other seasons only an causes peculiar colorations and on one occasional specimen will be seen. occasion we collected a quantity of bril¬ If specimens are located, collect a liant red hydras, so colored because of a quantity of the water plants to which
TURTOX Service Department Copyright, 1960, by GENERAL BIOLOGICAL SUPPLY HOUSE (INCORPORATED) 8200 South Hoyne Avenue Chicago 20, Illinois THE SIGN OF THE TURTOX PLEDGES ABSOLUTE SATISFACTION
Printed in U.S.A. Thousands of living Hydra in one of the Turtox laboratory tanks. they are attached and take them to the the most difficult problems connected laboratory in a pail of water. Place these with the rearing of Hydra is combating weeds in pans of water and the Hydra the periods of depression (see next page) will gradually expand so that they can be which affect them. When hydra become seen, and they may even come to the sur¬ depressed, the tentacles become shortened face and hang suspended from the under and the body contracted. They are un¬ side of the surface film. If a large glass able to eat and will soon die. A change aquarium tank is available, a quantity of of water will sometimes bring them out weeds may be placed in this aquarium of this state in a few days, providing the and the tank filled with water. After the water is taken from a culture in which specimens have expanded, they may be Hydra is flourishing. We have also found seen easily by looking through the aquar¬ that water taken from an aquarium in ium from the side. To obtain individual which plants are growing is very bene¬ specimens, clip off small portions of wa¬ ficial. This condition is most apt to ter plants to which Hydra are attached appear soon after the specimens are and place in another container of water. brought into the laboratory, and it may take considerable care to bring them How to Care for Hydra through the period. in the Laboratory. Hydra seems to do well in water which contains a slight amount of boiled aqua¬ If hydra are wanted only for the time tic plants. A very slight amount of fer¬ the class will be studying them, the living mentation seems to be beneficial if the specimens may be kept in a small-sized water remains perfectly clear. Water battrey jar or in a finger bowl full of thus prepared should stand for several pond or aquarium water. However, if the days before the Hydra are added. How¬ hydra are to be kept for a considerable ever, we have been equally successful in period a fairly large balanced aquarium rearing them in a small balanced aqua¬ containing growing water plants (but no rium of one-gallon capacity in which fish !) will be needed. Release the hydra were growing one plant of Vallisneria, in it and they soon attach themselves one of Sagittaria and one of Elodea. to the water plants and the glass sides of the tank. In handling living hydra two Hydra is a hearty eater and, if the important points must be kept in mind; culture is to do well, it must be fed first, water from most city water supplies plenty of Daphnia. This means that an will prove fatal, and they should there¬ active culture of Daphnia is necessary fore be provided with natural pond water when the Hydra are to be kept over or water taken from a balanced aqua¬ winter. rium; and second, rapid temperature changes are usually dangerous. Hydra Feeding. must be kept cool, for they can stand As has already been pointed out, Dahp¬ high (70°F) water temperatures only nia are an excellent food for hydra, but when the water temperature has in¬ any other small crustacean such as creased very gradually. Cyclops is acceptable. The small white- To grow Hydra successfully, two worm Enchytraeus albidus is also a good things are necessary—a good supply of food. It is impossible to keep living patience and plenty of Dahpnia. One of hydra in the laboratory for any great
(39-2) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
length of time unless a regular supply of they become nothing but stumps, this living food can be maintained for them. continuing until the hydra becomes a To demonstrate the feeding habits of small round ball and dies. During this Hydra to a group of student, pipette a period they refuse to eat and it was once few drops of water into a Syracuse thought that starvation was the cause of watch glass and place a Hydra in this. depression. Dr. I,. H. Hyman, of The Then add one living Daphnia or an en- American Museum of Natural History chytreid and the Hydra will soon en¬ suggests four possible reasons for de¬ tangle the organism in its tentacles, draw pression: (1) The temperature of the it into its oral cavity and consume it. water rising to more than 20°C. (2) insufficient oxygen in the water. (3) Ex¬ Reproduction. cess fermentation present. (4) Changing Hydra reproduces by means of buds of the hydra to water differing from that and by sex cells, but usually the two in which they were collected. types of reproduction occur at different In our own laboratory we have noticed times. When well fed, they will develop depression as a result of all of these buds very rapidly and in the course of conditions, but it also occurs at times a few days these buds become detached when no reason is apparent. from the parents and begin their inde¬ pendent existence as distinct individuals. Notes on Culturing Daphnia. Usually the sex organs (spermary and Since Daphnia are used so commonly ovary) are found on different animals, in feeding Hydra as well as aquarium but occasionally both will occur on the fishes it is well to keep a culture on hand. same individual. Fertilization and the A few suggestions are given to help those early development occur while the egg is interested. still attached to the parent. Hydra also If Daphnia are collected locally use produce winter eggs which are fertilized same water for culturing them as that in in the autumn and lie dormant until which they are found. Pond or aquarium spring. conditioned water should be used if the specimens are not collected locally. Depression. So far as we have been able to deter¬ Anyone who attempts to keep living mine by experimentation, there has been Hydra for a considerable period will no “sure-fire” method discovered of keep¬ sooner or later find that in spite of ing a culture of Daphnia permanently in plentiful food and other favorable con¬ the laboratory. Many methods have been ditions they will begin to die off. This is tried and some of them with a fair degree usually due to “depression,” the causes of success, but no perfect culture method of which are not clearly understood. De¬ which will work under all conditions has pression is that slow contraction often yet been found. The more successful found in hydras, where the body stalk methods use rather large containers— shortens and the tentacles contract until usually wooden tubs or small barrels
(39-3) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
having a capacity of from twenty-five to where Hydra are plentiful. fifty gallons. It is also well to keep the One of the easiest foods to rear in the cultures in subdued light and in a place laboratory for Hydra is the larva of the where the temperature is fairly constant. Brine Shrimp, Artemia salina. See Tur- The food varies greatly. Sheep manure, tox Service Leaflet No. 27. bone meal, living algae, lettuce leaves and boiled water weeds have been used References: successfully for varying periods. Morgan, “Filed Book of Ponds and Many other small fresh-water crusta¬ Streams.” ceans in addition to Daphnia are excel¬ Ward £ Whipple, “Fresh-water Biol¬ lent for feeding Hydra. Cyclops, Oypris, ogy.” small Gammarus and other similar forms are all good. One or several of these are See other useful references in Turtox usually found in abundance in waters Service Leaflet No. 14.
Materials for Study of Hydra LIVING MICROSCOPE SLIDES 3V21 Hydra oligactis (Pelmatohy- dra). These large Hydra are col¬ Z3.111 Hydra, extended specimen for lected in the Northern lakes and are general body structure, w.m.. $0.85 the largest species we have seen. They are ideal for laboratory study Z3.112 Hydra, adult with bud, w.m. and will be found much superior to 1.25 the smaller white Hydra. Available at all seasons. Bottle of 50. .$5.00 Z3.113 Hydra, male, showing gonads, Large. Per dozen 2.50 w.m 1.50 Large. Bottle of 25 3.50 9V11 Daphnia. Living Culture with Z3.114 Hydra, female, showing go¬ instructions 3.50 nads, w.m 2.50 9V1105 Brine Shrimp Eggs. Vial of about 50,000 viable eggs, vial of Z3.121 Hydra, x.s. showing detailed salt crystals for making the brine structure of ectoderm and endo- solution and cultural directions. derm 90 Price 1.75 Z3.122 Hydra, x.s. through male PRESERVED gonad 1.00 3X111 Hydra. Hydra oligactis, large size, preserved with tentacles well Z3.125 Hydra, x.s. through female extended. gonad - 1.10 Dozen 1.00 Hundred 8.00 Z3.131 Hydra, l.s., general structure. 90 3X112 Hydra. Large selected speci¬ mens showing budding. Z3.132 Hydra, l.s. through adult and Each 50 bud 1.25 Dozen 2.50 3X113 Hydra. Large, selected speci¬ Z3.137 Hydra smear preparations mens showing male sex organs. with nematocysts discharged and bud 1.20 Each 55 Dozen 5.50 3X114 Hydra. Large mature speci¬ mens showing female sex organs. Each 65 CHARTS Dozen 6.50 3X1141 Hydra Reproduction Set. In¬ Refer to your Turtox Biology Cata¬ cludes one of each (1) normal log for illustrations and descriptions specimens, (2) budding specimens. of Charts, Key Cards and Quiz Sheets (3) male and (4) female, in dealing with the structure and devel¬ separate vials 1.90 opment of Hydra.
All prices are f.o.b. our laboratories and are subject to change without notice
(3n-4) TURTOX SERVICE LEAFLET No. 16
THE CULTURE OF PLANARIA AND ITS USE IN REGENERATION EXPERIMENTS Living planaria are easily collected and kept in the laboratory and their remark¬ able powers of regeneration make them of unusual value in laboratory work. By following the simple steps outlined here, little difficulty will be experienced in keep¬ ing live specimens and demonstrating the phenomenon of regeneration. 1. Obtain living specimens by collect¬ ing in the field or by purchasing. Turtox can supply these in any quantity at all times of year. The smaller species, Planaria maculata, are easily collected in most localities. Look for them on the under sides of stones in pools or streams. The larger species, Planaria agilis or Planaria dorotocephala, are found in cold springs or spring-fed pools or streams, and can be captured by laying pieces of raw beef at the margins of the water. When the worms attach themselves to the meat they may be shaken into the collect¬ ing jar. 2. When the worms have been brought to the laboratory, place them in an enam¬ eled dishpan about half filled with clear water. (Pond water is better, but aerated tap water may be used.) Cut a piece of tin to serve as a cover to shut out the light. 3. Feed the worms every four or five days by placing in the pan small pieces of fresh calf or beef liver. It is inter¬ esting to note how quickly the worms can locafe the food. After two or three hours the worms will become gorged and leave the meat. At this time the food must be removed and the water changed, because Planaria cannot live in water which contains any decaying organic ma¬ terial. Agitate the water so that the l. Eye, S. brain, S. Auricle, k. Ventral nerve animals will drop to the bottom of the cord, 5. Intestine—anterior trunk, 6. Pharyn¬ geal chamber, 7. Pharynx, 8. Mouth, 9. In¬ pan, clean the organic material from the testine—posterior trunk. sides of the container and decant all the water. Pour on fresh water and, if not clear, repeat the process. Be sure cover with a cover glass. Gently flatten every bit of meat is removed. the animal, but do not exert too much pressure. Examine this mount under the General Laboratory Study low power of the microscope. To observe the general appearance and behavior place one or two live worms in a Regeneration Experiments watch glass containing a small quantity The flatworm shows remarkable powers of water and examine with the hand lens. of regeneration. If possible, give each To study the detailed structure place a student a few specimens, so that he can few grains of chloretone in the water. make several experiments. If material When the worm has become motionless, is not abundant, let several students work place it, dorsal side down, on a slide and together. Take several Planaria and cut
TURTOX Service Department Copyright, 1959, by GENERAL BIOLOGICAL SUPPLY HOUSE (INCORPORATED) 8200 South Hoyne Avenue Chicago 20, Illinois THE SIGN OF THE TURTOX PLEDGES ABSOLUTE SATISFACTION Printed In U.S.A. GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
Diagrams illustrating various cuts which may be made to demonstrate regeneration in Planaria. them into pieces, as indicated by the ac¬ companying diagrams. Planaria can be cut by allowing them to extend fully on a glass plate and then cutting with a sharp razor blade or Gil¬ lette knife. Experiment 1. Cut forward from the caudal end of the body, to a point just ahead of the mouth. Place specimen in a dish of water. Prevent the cut halves from growing together by recutting if the wound closes. Two tails should de¬ velop as a result of this experiment. Experiment 2. Remove head at “cut 1,” then make “cut 2” from the anterior end of the body posteriorly to a point just back of the mouth. The two halves should develop two heads. Prevent halves from fusing for a few hours as described in experiment 1. Living Planaria Experiment 3. Cut a 25-millimeter form postero-laterally. specimen into eight pieces. Place each Experiment 5. Remove head at “cut piece into a separate watch glass of water 1” and then cut a triangular piece out of and label so that it will be possible to the anterior third as illustrated in the tell from which section of the body each diagram. A normal head should develop piece came. The results of this experiment at “cut 1” and a very small head at the vary with the species used. side. Experiment 4. Make two diagonal cuts NOTE: Various abnormalities will oc¬ as indicated in the above diagram. The cur in these experiments. All of the re¬ center oblique section should develop a sults should be recorded by means of head antero-laterally and a tail should diagrammatic sketches.
Living Material Microscope Slides 5V11 Planaria (Dugesia). Large size. This is Z5.ll Planaria (Dugesia), w.m. Animal is the fresh-water planarian commonly used excellently fixed in an expanded condi¬ for experimental purposes. tion. The digestive system is completely in¬ Dozen $2.25 jected with carmine, in contrast with the Fifty 8.00 lightly hematoxylin-stained background of Hundred 15.00 the body $1.10
Z5.12 Planaria, x.s. through anterior, middle Preserved Material (pharynx) and posterior regions 1.35 5X15 Planaria (Dugesia). Large fresh-water planarian. 15 mm. or more in length. The Z5.13 Planaria. l.s. Each slide contains at large size and comparatively small amount least one section through the pharynx. 1.50 of pigment in the body make these speci¬ mens ideal for laboratory work. Flattened. Each 35 Z5.15 Planaria, w.m. of injected specimen Dozen 2.00 and uninjected specimen stained for cellular Hundred 14.00 structure. Both on one slide 1.60 All prices are F.O.B. our laboratories; subject to change without notice.
(16-2) TURTOX SERVICE LEAFLET No. 41
COLLECTION AND CULTURE OF EARTHWORMS AND OTHER ANNELIDS
The large earthworm, Lumbricus of the body out of the hole. By a quick terrestris, commonly used for dissection dart of the hand the worm is seized just in schools, is a somewhat delicate animal, where its body emerges from the hole not easily maintained under artificial and with a steady pull the posterior conditions in large concentrations. How¬ end is drawn out and the worm dropped ever, if large containers and the correct into the bucket. Should the pull be too soil mixture (see below) are provided, vigorous the collector feels a sudden and if at least one cubic foot of soil is giving way and finds himself holding a provided for each 40 earthworms, good half worm, the other half being securely results can be secured. If the earth¬ anchored in the hole. It is the collector’s worms are very large, it is often better hope that the rain will continue regard¬ to limit the population to 25 worms per less of its chilling effect and despite the cubic foot. fact that the ground over which he must Smaller earthworms (there are numer¬ crawl is becoming muddier and muddier, ous species) are hardier and can be for should the rain cease, the worms soon maintained in greater concentrations. retreat into their holes. Earthworms The manure worm, Eisenia foetida, is must be stalked with some care, for very hardy and easy to maintain in they are sensitive to the jar of the earth laboratory cultures. This worm is a good caused by the approaching collector and size for use in feeding toads, frogs, will jerk back into their holes like a snakes, lizards and aquarium fishes; and flash at the faintest vibration of broken it produces abundant cocoons from stick or incautiously placed hand, knee which very small worms emerge. or pail. Should the rain pour down in a steady, chilling sheet, the collector picks Collection worms with both hands, and, with con¬ tinuous rain, many of the worms leave Earthworms (Lumbricus terrestris) their holes entirely. Copulating pairs may be collected in quantity on rainy are now numerous and two worms can nights during the spring and fall. In be scooped up with a single motion. summer the rains are likely to be short Should the rain continue all or most of and hard followed by quick drying, the night and into the next morning, the giving the worms little opportunity to streets and roads may become covered emerge. The best collecting season is in with worms that have crawled or been the spring. During the winter the worms washed onto the hard surface and are are confined below the frozen earth, but unable to escape by burrowing. with the spring rains and the thawing of this frozen crust they swarm to the Culture surface. In the Chicago region this In many parts of the United States swarming usually occurs during the earthworms of one or more species are months of April and May, at which time plentiful and small quantities can be spells of rainy weather of two to three collected during the spring, summer or or more days duration are common, with autumn by digging for them in gardens, the precipitation heaviest during the barnyards or moist woodlands. If large night. It is during the time of these cold spring rains that the earthworm collector numbers of worms are required, outdoor beds can be established, or large indoor equipped with rubber boots, raincoat, containers can be maintained. bucket and flashlight, repairs to the lawns and shrubbery. On hands and A very satisfactory and productive knees he moves cautiously, and with outdoor “earthworm farm” can be made straining eyes endeavors to distinguish as follows: the worms from the twigs and leaves 1. Select a shady place where the soil with which the ground is covered. As is well drained and excavate an area visual adjustment comes, the worms about 6 by 6 feet (or larger if desired) may be seen lying with the anterior half and about three feet deep. If the under-
TURTOX Service Department TURTQr^BUCTS Copyright, 1960, by GENERAL BIOLOGICAL SUPPLY HOUSE (Incorporated) 8200 South Hoyne Avenue Chicago 20, Illinois
THE SION OF THE TURTOX PLEDGES ABSOLUTE SATISFACTION Printed in U.S.A. GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS lying soil is heavy clay, line the ex¬ coons can usually be found at all cavated area with six or eight inches of seasons. coarse sand and gravel. Enchytrae Worms 2. Fill the reminder of the bed with a mixture of equal parts of light loam, Every fish fancier knows “Enchytrae” well rotted manure and leaf mold. The as one of the best foods for his fish. leaf mold should contain a good propor¬ Equally important is it for food for tion of half-rotted leaves. fish in laboratory aquaria and for the small salamanders such as red-spotted 3. After this has settled, wet it newts. The scientific name of this thoroughly and place the worms on the “domesticated” annelid is Enchytraeus surface of the bed. albidus. If the surrounding soil is heavy clay, The cultural methods for this form sand or rather barren and dry, the worms are quite similar to those of earthworms. will remain pretty well within the bed, Secure a tight wooden box and fill it up breeding and increasing over a period to within two inches of the top with rich of years. However, if the surrounding humus, sifted to remove coarse particles. soil is a rich, light loam, the worms are Place the original culture in a hole in likely to spread out and migrate slowly the center. Cover it over with humus and to surrounding areas. This can be pre¬ then cover the box with a wooden or glass vented, at considerable cost, by lining the cover to prevent it from becoming dry. excavated area with a wooden framework Place the box in a cool basement, the covered with fine-meshed copper or temperature should not become higher bronze screening. than from 60° to 65 °F. Add water It is well to introduce small quantities sparingly—just enough to keep the of well-rotted manure and rotted leaves humus damp. into the bed once or twice a year, and Feed Enchytrae bread soaked in milk to keep the surface moist and covered by working small holes in the humus, at all times with a layer of leaf mold filling them with the food and covering several inches deep. In the northern over with an inch of humus. Alternate states, a covering of a foot or so of foods are cooked oatmeal, mashed straw, weighted down with poles or potatoes, mashed potatoes mixed with boards, will retard frost to some extent rich chicken broth, and the like. Feed and make the living worms available worms sparingly about twice a week, during the winter months. but never until previous food has been consumed. If food sours, remove at once. Containers for maintaining earthworms indoors are more convenient than an For freeing the worms of dirt when outdoor bed if large quantities of worms ready to feed them to laboratory animals, are not required. The essentials of indoor place a quantity of them, dirt and all, earthworm culture are (1) a large con¬ in a glass jar or paper cup and fill tainer, 4 to 6 cubic feet minimum; (2) it one-quarter full of water. Within 30 a temperature lower than 60° F; and minutes, the worms will have crawled (3) proper control of moisture. Earth¬ up the sides, where they may be secured worms cannot be crowded and not more free of all dirt. than 40 medium or large specimens should occupy one cubic foot. A con¬ Aquatic Earthworms venient and practical container is a Tubifex and other aquatic earthworms wooden box measuring 4x4x2 feet; or are found in the mud at the bottom and an oak barrel or large keg can be used. along the shores of most bodies of fresh For best results the temperature must water. They can be seen waving aimlessly be kept between 40° F. and 55° F. The above the mud and when disturbed container should be filled with a mixture retreat into their slime and mud tube. of very light loam (never use clay), Some species live in decaying vegetation partly rotted leaves, leaf mold and well and others are found in considerable rotted cow or horse manure. This mixture numbers in floating masses of algae. should be kept slightly moist (but never wet or “soggy”) at all times. To retard Aquatic earthworms are collected with evaporation and help maintain an even the mud or decaying debris in which temperature, the container can be they live and then washed free of the covered with a plate of glass or a piece mud by placing the collected material in of well varnished (to prevent warping) cheese cloth or fine-screened sieves. plywood or wallboard. If the soil contains It is easy to keep a culture of these plenty of dead leaves, no special feeding worms growing in an aquarium. A layer will be necessary. of mud about an inch deep, containing In a container of this type, one can Tubifex or other aquatic earthworms is keep considerable quantities of adult placed on the bottom of the tank and worms, and some egg capsules or co¬ covered with a half inch layer of sand. (41-2) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
A pair of copulating earthworms (Photographed late on a rainy night)
The tank is prepared as a balanced Hirudo medicinalis is still used in medi¬ aquarium with plants and animals. After cine to a small extent. This practice was the water has cleared the Tubifex will so prevalent in Europe many years ago be seen, their bodies extending above the that “leech” became practically synony¬ sandy layer. Jarring the aquarium will mous with “physician.” cause them to retreat momentarily. It is Care in the Laboratory. Each year, Tur- wise to keep fish out of the tank in which tox imports large numbers of the medici¬ Tubifex and other aquatic earthworms nal leeches from France. They usually are kept. reach us packed in damp earth. In fact, Aquatic earthworms are good fish food. they can live in this condition for rela¬ To obtain them in quantity free of mud tively long periods of time. For actual they should either be washed clean as study, it is much better to keep them stated above or the mud containing the in balanced fresh water aquaria. Care Tubifex and other species should be should be taken that the aquarium is placed in shallow pans over a warm well supplied with the large oxygenating radiator or low flame. The warmth plants such as Elodea, and Vallisneria. drives the worms to the surface. Place the aquarium so that it receives very little direct sunlight as leeches pre¬ fer darkness. Provide stones, leaves and Leeches other opaque objects under which they may hide to escape the light. Leeches are to be found in almost every The usual animal population of the standing pond, ditch, lake, and sluggish aquarium, such as small fish (bullheads, stream. Many of them are beautifully sunfish, and the like), tadpoles, small colored and add much attractiveness to crayfish, salamanders, snails, and insect the ordinary balanced fresh water aquar¬ larvae, should be kept in the aquarium. ium. Their care is a simple matter and The excess food given the fish promotes yet their behavior is of great interest. growth of the small forms, which in turn Although a number of leeches are to be furnish some of the food of the leeches. found in local ponds, the one most com¬ Most leeches will partake of blood at monly studied in the laboratory, both some time during their life. Some are preserved and living, is the imported parasitic upon turtles, frogs, salaman¬ medicinal leech, Ilirndo mediciualis. Of ders, and fish. Although it isn’t essential particular interest is the giant leech, that they have blood to live for only a Haemopis grandis, the largest American month or two, it is interesting to know species, which is now much used in school just what kinds of blood they prefer. laboratories. The turtle is frequently used to supply
(41-3' GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS the blood for live leeches (the leeches maintaining them for experimental pur¬ are placed upon the soft skin of the tur¬ poses if but slight attention is given. tle). Rabbits may also be used for this purpose by shaving the hair from a small area of the ventral skin. Leeches will Marine Annelids sometimes feed upon fresh liver if placed Students living near the seacoast can In a fingerbowl along with it. These and collect living marine worms such as other methods determined by experimen¬ Nereis, Amphitrite, and Arenicola. Some tation will be helpful when leeches must of these will live for several weeks in a be kept for long periods of time. Leeches salt-water aquarium, and are far more have lived in our aquaria, practically interesting than preserved specimens. Without attention, for as long as a year. Refer to Turtox Service Leaflet No. 20, Teachers will experience no difficulty in Notes on Marine Aquaria.
MATERIALS FOR THE STUDY OF ANNELIDS 8V11 Lumbricus. Earthworms. Large 8V12 Enchytrae. A small, thread-like, mature specimens, each with clitel- white worm about 25 to 30 mm. in lum. Usually available at all sea¬ length, especially useful as a food sons, but must be ordered two weeks for small aquarium animals. Instruc¬ in advance of delivery date. tions for the care of the living Dozen $3.00 Enchytrae are sent with each ship¬ Hundred 14.00 ment. Large portion, sufficient to start a permanent “culture”. . .$3.50 8V112 Earthworms. Small specimens, suitable as food for snakes, sala¬ manders, turtles and the larger 8V14 Tubifex. Small aquatic earth¬ aquarium fishes. Order two weeks worms found in pond mud. Large in advance. Lot of fifty with direc¬ portion with instructions for start¬ tions for establishing a permanent ing permanent culture 4.00 culture 4.00 Per hundred 7.00 8X264 Lumbricus. Preserved Earth¬ 8V115 Earthworm Cocoons. The co¬ worms. Large size, 9" to 12" for dis¬ coons or capsules contain eggs or section and laboratory study. minute earthworms in various stages Dozen 2.10 of development. Easily kept in moist Hundred 16.00 filter paper for study; culture and instruction sheet sent with each shipment. Available from November 8X266 Lumbricus. Preserved Earth¬ to May. worms. Mature specimens, size 7" Per dozen 1.25 to 9". Dozen 1.70 Per hundred 6.00 Hundred 12.00
The special 64-page catalog, Turtox Dependable Biological Supplies lists all of the material needed in beginning courses in Biology, Botany and Zoology. This catalog is especially helpful in planning requisitions for purchases under Title III of the National Defense Education Act.
A comprehensive selection of Turtox Charts and Turtox Key Cards of Earthworm, Leech and other Annelids is available. Write for the free Turtox Three-Way Checklist of charts and biological drawings.
All prices are f.o.b. our laboratories and are subject to change without notice.
(41-4) TURTOX SERVICE LEAFLET No. 27
BRINE SHRIMP AND OTHER CRUSTACEANS This leaflet discusses very briefly the debris attached to them) are placed. care of the brine shrimp, Artemia salina, For quicker results scrape some of the and some of the fresh-water crustaceans green slime from the glass side of a commonly studied in school laboratories. fresh-water aquarium and place a little For information on barnacles and ma¬ of this in the brine shrimp culture. rine crabs, refer to Turtox Service Leaf¬ (Many of these unicellular fresh-water let No. 20, “Notes on Marine Aquaria.” algae will grow in a weak brine solution as readily as they grow in fresh water.) Brine Shrimp Use as Food. For feeding Hydra, the The brine shrimp, Artemia salina, is a larval brine shrimp may be concentrated phyllopod crustacean of very general by straining the culture solution through distribution. It is extremely interesting a piece of fine-meshed cloth. They are as a marine crustacean which can be then placed in the fresh-water tank con¬ easily reared and studied in the school taining the Hydra, where they will live laboratory, and it has great practical for several hours, usually long enough to value as a constant source of living food provide the Hydra with ample food. Be¬ for Hydra and small aquarium fishes. cause of the very low cost of the eggs The resting eggs of the brine shrimp and the great rapidity with which they float in the water and require drying hatch, new cultures can be started every before they will hatch. This feature few days if a constant food supply is makes them available at all seasons, for wanted. the dried eggs remain viable (alive) for The earliest larval stages are also very several years if kept in a dry and fairly useful as food for small aquarium fishes; cool place. or the brine shrimp may be allowed to Hatching the Eggs. The eggs hatch develop and grow larger if they are de¬ very quickly (within 24 to 48 hours) sired as a living food for larger fishes. after being placed in a brine solution Laboratory Study. Be sure to examine which is kept at a temperature of 70° the eggs (both dry and hatching) and to 75° F. Natural or artificial sea-water the nauplius larvae under the micro¬ may be used if available. However, the scope. It is a most interesting study and brine solution used as a hatching medium it permits you to show your students may be almost any concentration from the continuous development of living about 0.1% to 6%. When sea-water is examples of a marine crustacean. not available make up a brine solution by adding two teaspoonfuls of common Glass Shrimp table salt to one quart of water. After This fresh-water shrimp, Palaemonetes hatching, the larval stages (nauplii) may exilipes, or “glass shrimp”, as it is often be used as a food (see below) or may be called because of its remarkably trans¬ transferred to other containers for fur¬ parent body, is an interesting crustacean ther development. If thousands are al¬ for laboratory study, and a valuable lowed to remain in one container they scavenger for the balanced aquarium. It will die after a few days for lack of lives well in either a small or a large food and sufficient oxygen. balanced aquarium, but prefers a tank Rearing the Shrimp. If you wish to containing plenty of vegetation. It lives rear Artemia to maturity and establish harmoniously with most small fishes, but a permanent culture, place only a few must not be placed in an aquarium con¬ of the larvae in a quart or larger con¬ taining fishes large enough to eat it. tainer of brine solution and supply them Palaemonetes will sometimes breed if a with food. Yeast is a satisfactory food, considerable number of individuals are as are also various of the one-celled kept in a large aquarium, and the fe¬ floating algae. The latter will usually males carrying eggs are particularly in¬ appear in a culture of natural sea water teresting for study. When hatched, the rather soon and will eventually appear young shrimps are very small and will in any brine solution in which the brine form food for even the smaller fishes un¬ shrimp eggs (together with the natural less they are immediately transferred to
TURTOX Service Department Copyright, 1959, by GENERAL BIOLOGICAL SUPPLY HOUSE ( IX CORPORATED ) 8200 South Hoyne Avenue Chicago 20, Illinois THE SION OF THE TURTOX PLEDGES ABSOLUTE SATISFACTION Printed in U.S.A. GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
Larva of the Brine Shrimp. a tank where this danger does not exist. shrimps again swim about. The glass shrimp is largely a scaven¬ Eubranchipus when adult is about one ger in its feeding habits and is therefore inch in length and reddish or bronze in useful in keeping the aquarium clean. It general coloration. The females are will eat particles of fish food and may larger and usually much more plentiful occasionally be fed tiny pieces of oyster, than the males. All species of fairy raw fish or raw meat. shrimps swim on their backs, propelling themselves rather gracefully by means of Fairy Shrimp their eleven pairs of waving gill-feet. During the late winter or early spring, They feed upon diatoms, protozoans and it is not uncommon to find small pools other microscopic forms. containing thousands of fairy shrimps. We have never been able to discover All disappear after a few weeks and any dependable way of maintaining per¬ the pools in which they lived, being of a manent laboratory cultures of Eubran¬ temporary type, may dry up completely chipus, although dormant eggs brought during the summer months. Not so many into the laboratory and placed in water years ago, the fairy shrimp was pointed will usually hatch at any season. Speci¬ to as a splendid proof of the theory of mens collected in the spring will usually “spontaneous generation”; it appeared live for several weeks in jars or aquar¬ in unbelievable numbers in pools formed ium tanks. The temperature of the by the melting snow, it disappeared as water should be kept as low as possible, suddenly as it had come and then in a as high temperatures hasten their devel¬ few weeks the pool in which it had lived opment and shorten their life span. became a dry and dusty bit of ground. The explanation, of course, as every Daphnia teacher now knows, is that this creature Daphnia (or “water-fleas”) of various produces resting eggs which sink to the species can be collected in small quanti¬ bottom and live in a dormant stage for ties ordinarily by swishing a fine-meshed long periods. Indeed, some of these eggs net among the aquatic plants growing in appear to require drying before they can shallow water. At some seasons, often develop, and all fairy shrimp eggs can go during the autumn, Daphnia and other through extended periods (several years) small crustaceans occur in great abun¬ of drought and freezing without harm. dance in the shallow waters of stagnant When conditions are again suitable ponds and ditches. Since Daphnia are (when the spring thaws flood the pools), used so commonly in feeding hydras and they hatch and thousands of fairy small aquarium fishes, it is well to keep
(27-2) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
Female Cyclops with egg sacs.
cultures of them in the school laboratory. decayed aquatic plants. Keep the cul¬ If Daphnia are collected locally, cul¬ ture in subdued light and in a place ture them in the water in which they where the temperature is fairly con¬ are found. If the specimens are not stant at'65° to 70° F. obtained locally, use pond water or wa¬ ter from an established and well bal¬ Other Fresh-Water Crustaceans anced aquarium. Cultures of mixed crustaceans may It is only fair to admit that we know include many small forms which occur of no “sure-fire” method of maintaining in ponds and slow-flowing streams. The permanent cultures of Daphnia. Many cultures which we furnish during the methods have been tried and some of autumn and winter months usually con¬ them with a good degree of success, but tain Daphnia, Cyclops, Cypris, Latona, no perfect culture method which will Gammarrus and others. Most of these work under all conditions has yet been will live well in cultures or in small bal¬ discovered. The more successful methods anced aquaria. use rather large containers — usually A small aquarium of from one to five wooden tubs or barrels having a capac¬ gallons capacity is suitable for mixed ity of from twenty-five to one hundred crustaceans. Such an aquarium should gallons. Four or five cultures should be contain sand and growing plants and maintained, for then there is a good some bottom debris (dead leaves, mud, chance that at least one culture will con¬ etc.) from a pond. It can contain a few tain numerous Daphnia at any given time. snails and small clams, but must not Professor Harold Heath of Stanford include fish which would feed upon the University recommends the following crustaceans. method: “Use one ounce (by weight) of Gammarus will live well and repro¬ dried sheep manure to one gallon of duce abundantly in a small tank con¬ water. A wooden trough holding twenty- taining a few dead leaves and other five gallons of water is used as a con¬ decaying vegetable matter. These smaller tainer. After the sheep manure and crustaceans need little attention and water have been in the trough for two will continue in fair numbers in any or three days, the Daphnia are added. small tank containing pond debris and A few lettuce leaves are placed in the some algal growth. culture from time to time and are re¬ placed by fresh leaves as they become Crayfish thoroughly decayed.” Crayfish are of very general distribut- Other foods frequently used include tion and adult specimens can usually be bone meal, decayed masses of algae and obtained in lakes, ponds, and streams
(27-3) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
Living Daphnia photographed in a culture.
without difficulty. Being to some extent The crayfish is one of the most inter¬ nocturnal, they hide during the day and esting forms to keep in the laboratory will be found by lifting up flat stones or and two or three should be included in other objects under which they retreat a large aquarium. Students will learn for protection. In places where cray¬ more of the habits of this animal by ob¬ fish are plentiful it is usually possible to serving a few living specimens than they secure small specimens by swishing a will by reading chapters of textbook net around among the aquatic plants or material. An adult may be placed in a by dipping up netfuls of bottom debris. gallon battery jar containing clear wa¬ Crayfish which are to be kept in aquaria ter and the students can observe the in the laboratory should be collected movements of the mouth parts, antennae from ponds or quiet rivers, as those and appendages. Feed the specimen a taken from fast-flowing streams are less small piece of raw fish or beef and ob¬ likely to live in captivity. serve its method of taking food. In the late autumn or early spring Crayfish often uproot or otherwise dis¬ look for the females carrying eggs at¬ turb the plants in an aquarium unless tached to their swimmerets. If one is they are provided with stones under found, take her to the laboratory, place which they may retreat. Therefore, it her in an aquarium and watch the eggs is a good plan to have a loose pile of develop into young crayfish. Often fe¬ good-sized stones in one corner of the males carrying young will be found in aquarium, and also to select small cray¬ the early spring, although within a few fish as aquarium inhabitants. Two or days the fully developed young will leave three specimens measuring about two their mother’s swimmerets to shift for inches or less in length are enough for themselves. a six gallon tank.
Refer to your Turtox Biology Catalog for current listings and prices of living crustaceans. Brine shrimp eggs are available as follows. 9V1105 Brine Shrimp Eggs. Vial of about 50,000 viable eggs of the brine shrimp, Artemia salina, a vial of salt crystals for making the brine solution and complete directions for culturing. These eggs are available at all seasons and will hatch within two days after being placed in the brine solution. Excellent for the living study of the development of a marine crustacean and as a food for Hydra and both fresh-water and marine aquarium fishes. Complete culture set as described.. .$1.75 All prices are f.o.b. our laboratories and are subject to change without notice.
(27-4) TURTOX SERVICE LEAFLET No. 7
THE CARE OF FROGS AND OTHER AMPHIBIANS
Frogs times. Keep it dark and do not disturb Grassfrogs (Rana pipieus and others). the frogs often. Inspect at least every One of the most frequent inquiries reach¬ third day (oftener if there has been a ing tile Turtox Service Department is disease) and remove any frogs that have the one which asks, “How can I keep died or are in a weakened condition. If living grassfrogs in my laboratory?” disease should become rampant, sort The answer is not as simple as one might over all specimens and segregate those suspect, for different methods are used, not in the best condition. Flush out the depending on the number of frogs to be tank thoroughly, remove specimens to a kept, Uie length of time they are to be temporary storage receptacle and ster¬ maintained, and the time of year the ilize the tank with a strong salt solution, stock is purchased or collected. a formalin solution or lime. Return the The small laboratory may wish to frogs only after a thorough washing out keep only four to six specimens for a of the disinfectant. Always disinfect relatively long lime. A large woodland tanks thoroughly after each lot of frogs terrarium would be almost ideal, if there is used and also just before another were a sunken dish in it in which water is batcli is to be introduced. kept. Or, the semi-aquatic terrarium In such concentrations, it is seldom can be used with success. In either case, advisable to feed the frogs, as it would you will have provided the approximate be an unending task. If kept cool, un¬ environment afforded in nature. These disturbed, and dark, there will be but are meadow frogs, frequenting almost little activity and, hence, but little dry fields in the summer in quest of in¬ necessity for food. During the winter sect food, and returning to the ponds months the frogs can stand much more for hibernation and subsequent egg-lay¬ water and the tank described above may ing the following spring. Feed them be altered so as to keep a constant level living insects such as mealworms, cock¬ of water about one to two inches deep. roaches, small grasshoppers if available, Another method for maintaining liv¬ Hies, caterpillars, and the like. They ing frogs consists of a tank, (or any ean sometimes be trained to accept beef other suitable container) provided with liver, lean beef, etc., moved before them a layer of damp (not wet) sphagnum on the end of a string or broom straw. moss. (The dry baled moss used by See that eacli individual receives some florists is suitable.) The slight amount food. of iodine in the moss appears to prevent Some of the laboratories of our large red-leg and other infections. The universities wish to keep great numbers sphagnum moss tank should contain of frogs available for several weeks. only the damp moss—no water. Their care is often a real problem. We Care of Frogs upon Arrival. Just as recommend large wooden tanks (cypress soon as you have received your frogs, is perhaps most economical in the long whether there be three or three hundred, run) with a gradual slope of pebbles, open the package carefully and then about %" diameter, in one end. The inspect each one to be sure it has not tank is to be equipped with an inlet so suffered from its long journey. It is that water is constantly trickling well to have a pail of fresh water handy through it, and an outlet which drains and to wash the animals off before plac¬ all but about %" of water in the bottom. ing them in your tanks. Any frog which Not much water is needed, especially appears to be diseased should be when the frogs are kept during the segregated. summer. That which flows through the tank tends to keep it clean and free Bullfrogs and other aquatic frogs. from wastes. However, flush it out Bullfrogs may be collected in swamps thoroughly every four or five days. It is and lakes and they occur more or less best to locate such a tank in the base¬ throughout all the eastern half of the ment, where it will remain cool at all United States. However, most bull-
TURTOX Service Department Copyright, 1960, by TURTOXmOIUCTS GENERAL BIOLOGICAL SUPPLY HOUSE (IXCORPOEATED) 8200 South Hoyne Avenue Chicago 20, Illinois THE SIGN OF THE TURTOX PLEDGES ABSOLUTE SATISFACTION
Printed in U.S.A. GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS frogs sold by dealers and supply houses cover. In a terrarium of this kind the come from the Gulf States, as these humidity is easily controlled and the southern frogs are larger and bullfrogs tree frogs are protected from drafts and are much more plentiful in the South. sudden temperature changes. The ter¬ It is not uncommon for them to be rather rarium should contain loam and leaf sluggish when they arrive at northern mold and may be planted with mosses, destinations during the winter and spring ferns, liverworts and other woodland months. They will become lively as soon plants. as they are placed in water and allowed Tree frogs require living food and to warm up gradually, say, to about will readily accept almost any small in¬ 76“ F. sects. They will occasionally take Upon arrival, the bullfrogs should be worms and will sometimes accept tiny washed in clear water to free their bodies pieces of raw meat presented on the end of any wastes which might have ac¬ of a toothpick. During the winter cumulated during shipment. Place them months living Drosophila (fruit flies) then in a large aquarium; probably one and cockroaches (both of which are equipped with running water would be easily reared in laboratory cultures) are best. It should be screened over to quite satisfactory as food. Small meal¬ prevent the frogs from leaping out and worms may also be used. dying when no one is in the laboratory. These animals can float on the water for long periods of time but should be pro¬ Frog Eggs vided with rocks or other supports upon which they can rest. If large quantities The eggs of the grassfrog, Rana of bullfrogs are to be kept, the wooden pipiens, are found in ponds and ditches tanks recommended in the section deal¬ in early spring. Eggs of the spring ing with grassfrogs are very desirable. peeper, Ilyla crucifer, appear at the same time, but toad eggs are not found Feed bullfrogs upon small crayfish, until the first part of May, and bullfrog minnows, earthworms, young grassfrogs eggs are usually not found until June. and the like. Green frogs may be fed earthworms, mealworms and flies. It is It is interesting to follow the devel¬ possible to train either of these frogs opment of the frog from the egg, to take food to which it is unaccustomed through the tadpole stage, to the adult in nature—lean beef, beef liver and frog. Any makeshift aquarium can be even canned shrimp if it is dangled used for this study and since many before the animal so that it appears In schools have a fresh-water aquarium we have motion. suggest placing a few eggs in it for ob¬ servation. Green frogs (Rana clamitans and other Frog eggs are found in shallow water. aquatic species) require about the same When they are collected they should be care as bullfrogs. Being much smaller, brought to the laboratory in pails of the however, a few individuals will live nice¬ pond water in which they are found. ly in a swamp or semi-aquatic terrarium. In the laboratory the eggs may be placed in shallow pans, or a small cluster can be added to the fresh-water aquarium. Tree Frogs It is best not to place an entire clump of frog eggs in the tank lest there be more Tree frogs, or tree “toads” (ITyla) as tadpoles in the aquarium than the water they are commonly called, are plentiful can support. Approximately twenty-five in many sections of the United States, to fifty eggs are enough for a demonstra¬ but because of their secretive habits and tion. Small lots of eggs can be kept in protective coloration they are seldom finger bowls, provided that the water is seen by the casual observer. Tree frogs changed frequently. Large masses of live in moist situations, spending much of eggs can be kept in dishpans which are their time in trees and hushes and in artificially aerated until the eggs have the case of most species, resorting to hatched, but then they must be spread ponds only during the breeding season. around in other containers or better still They live remarkably well in a school the majority of tadpoles should be re¬ terrarium and are interesting amphibi¬ turned to the pond where they have a ans for laboratory observation. much better chance for survival. Tree frogs must be maintained in a The rate of development of the frog humid situation where sufficient moisture egg is determined almost entirely by is available at all times; the woodland the temperature of the water. A temper¬ type of terrarium is well suited to them. ature range of 50“ to 60“ F. is satisfac¬ (Note: See Turtox Service Leaflet No. 10, tory, but a water temperature of 70“ “The School Terrarium.”) Screened ter¬ to 75“ will cause more rapid develop¬ raria are not satisfactory; the best type ment especially where the -food supply is a terrarium housed in a standard is adequate. Overheating of the water rectangular aquarium tank with a glass will cause the eggs to die. (7-2) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
At a temperature of 70° to 75° the prevalent in your locality and since all young tadpoles will attain swimming size have much in common it will be a simple in about two days. Watch the egg mass matter to vary our suggestions to care closely and should it turn white, it is for the species in which you may be an indication that the eggs are unfer¬ interested. tilized or dead. The whole mass will pol- 'ute the water and should be discarded. Healthy eggs soon turn dark all over Newts. The common red-spotted newt as they develop. receives our first attention, because, without a doubt it is the one most gen¬ erally seen in the laboratory. There are two distinct phases in the life of this Toads animal; the red eft or land phase and The common toads do well in certain the olive green, red spotted aquatic terraria. Size is an important matter phase. Tlie red eft is an ideal woodland in keeping them; the small to medium- terrarium animal; its brilliant color con¬ sized animals usually being sought as trasting so vividly with that of the moss- they adapt themselves better to the covered terrarium floor. This phase does small area afforded in a terrarium. Too, nut require the aquatic habitat and a toad will burrow into the soil and, the should never be placed in an aquarium. larger he is, the more trouble one will It feeds upon very small insects, young encounter in keeping plants growing spiders, small ants and similar living on the soil surface of the terrarium. food. White worms (Enchytraeus) are Terraria are planted in the usual way relished. Bits of lean beef and calf liver with some soil in the bottom covered can be fed with forceps. The animals over with mosses, lichens, and the like. being very small, require but infinites¬ Ferns, the dwarf varieties, are planted imal amounts of food. in one end. A group of rocks may be The aquatic phase of the newt should arranged in the opposite end and, if be kept in an aquarium. It is well if there are some cracks, in which the toad the tank is heavily planted so that the may hide, perhaps he will take to them leaves form mats in some places on the rather than resort to burrowing. Do not surface of the water, as the newts like disturb the animal for quite some time to crawl out upon them at will. Feed after establishing it in its new home. this animal by removing it from the tank Provide a shallow drinking pan. to a lingerbowl of tepid water into Feed them living specimens at first— which has been dispersed small particles cockroaches, caterpillars, mealworms, of lean beef or liver. Leave it here until Hies if available, ants and young spiders. it has eaten as much us it likes, then Later, you may have the toad take sev¬ wash it off and return it to the aquarium. eral angle worms or even lean beef held About once a week is satisfactory for in front of it at the end of a toothpick feeding unless the animal has a voracious or broom straw. There is no danger of appetite, in which case you may feed over-feeding, although it is not neces¬ as often as it appears to lie hungry. sary to feed every day. Two or three Once the animal lias become accustomed times per week is generally sufficient. to a regular feeding schedule and to this type of food, one may be successful in In small terraria, it is not advisable feeding it in the aquarium. The only to keep salamanders with toads. The danger is that food particles introduced poison from the toad skin is quite might not be eaten and then decay, thus detrimental to other small amphibia. fouling the aquarium. This point cannot Toads must be kept warm if they are to be over-emphasized in feeding any lie active. When room temperatures aquarium inhabitants—excess food is fall below 70° F., toads usually burrow very apt to spoil the entire set-up. under and may not be seen for long periods. An electric light bulb suspended above the terrarium frequently has a The Large Adult Salamanders. The tiger Deneficial effect upon them. They may salamander (Ambystoma tiyrinum), the not come out into the bright light, but spotted (A mby.stoma maculatum) and it warms them up so that, as soon as the marbled (A. opacum) are com¬ the light is removed, they will usually monly in stock in the proper season in hop out of their retreat and feed. the Turtox Laboratories. They make good terrarium inhabitants, thriving in either the woodland or semi-aquatic habitais. They frequently burrow beneath Salamanders I he moss covering to hide but are less There are many kinds of salamanders likely to do so if some crevices between which may be kept in the laboratory and rocks or small “logs” are provided. This we give here brief directions on the care type of environment is also suitable to of those which are more generally main¬ the giant newt of the Pacific Coast, tained. Other salamanders may be Trilurus tornsus. They feed upon meal- (7-3) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
The Tiger Salamander, Ambystoma tigrinum. worms and other small insects but one must be given special attention. Their can train them to accept, quite willingly, care approximates that indicated above large amounts of hamburger. They have for the red eft. to be fed with forceps at first but even¬ tually will feed out of a small dish. Feed Necturus, Cryptobranchus and Amphi- at regular times and less trouble will be uma. These large amphibia are not generally suitable for balanced aquaria encountered. Vary the diet with liver and (unless they be of small size and the experiment with various other raw meats. aquarium of large capacity) and are Triturus pyrrhogaster. This form (the most successfully maintained in the run¬ red-bellied salamander) lives well in the ning-water aquarium. They may be kept ordinary balanced aquarium and may for long times in such tanks, even in be cared for in the same manner as large wooden tanks where the water described above for the aquatic phase of depth is maintained at a constant level the red-spotted newt. of about four inches (a continuous flow being provided). They will eat earth¬ Plethodons. These small salamanders worms, minnows, small crayfish, large make good terrarium animals, preferring water bugs and calf liver. Do not allow the moist but not wet woodland environ¬ the water to become warm during the ment. They are delicate and therefore summer months.
Additional Turtox Service Leaflets offering information on the care of Amphibians are: No. 10 The School Terrarium No. 23 Feeding Aquarium and Terrarium Animals
13V25 Triturus viridescens. The Red-spotted Newt, aquatic phase. Two for $1.35 Dozen 7.50 13VS3 Tree Frogs. Will live well in a moist woodland terrarium. Three for 3.50 Dozen 10.50 Write for current prices of grassfrogs, bullfrogs, Ambystoma, Necturus and other amphibians. All prices are f.o.b. our laboratories and are subject to change without notice.
(7-4) TURTOX SERVICE LEAFLET No. 40
THE CARE OF RATS, MICE AND GUINEA PIGS Rats: one-hole rubber stopper and a short The albino rat has long been used in piece of glass tubing. One end of the anatomical, physiological and psycho¬ tubing is sealed in a flame. Continue logical experimental work, and because to apply heat to it uniformly while so many data are available it is con¬ blowing through the other end so as to sidered a standard laboratory animal. produce a bulbous enlargement in the A pair of rats may be kept in a small heated portion. This bulb will burst, cage, measuring ten inches in height, leaving a small opening which is flamed. ten inches in diameter, and constructed Heat the tube about one inch away from of galvanized hardware cloth of three- the enlargement and bend it slightly. In¬ eighths inch mesh for top and sides. The sert the glass tube into the one-hole rub¬ bottom should be made of one-half inch ber stopper which is then inserted into mesh to allow the feces to drop through. either the bottle or test tube. The filled The top of the cage should be removable fountain is inverted and attached to the or hinged, to facilitate removing the outside of the cage. Be careful that the animals for observation and for clean¬ glass tip protrudes slightly through the ing the cage. A shallow pan should be open mesh into the interior. It should be placed underneath the cage to catch the at convenient height for the animals. urine and fecal matter. This pan must Food and water are of primary im¬ be cleaned daily to avoid any undesir¬ portance in the growth and well-being able odors in the laboratory. of all laboratory animals. The stomach If more than one pair of animals are of the rat is so small that it cannot to be kept, a larger size cage is required. take in enough water to last for a long It should be constructed of the same time. Therefore, an abundant water sup¬ material, but it should be nineteen inches ply should be available. long, twelve inches wide and nine inches Because the rat is an omnivorous high. The cage also should be equipped creature, its food requirements are pro¬ with a pan, and the top hinged, or a vided easily. Table scraps are a very long door made on one side. Larger satisfactory food. A well balanced diet cages than the above size may be used, containing all the elements necessary but they are difficult to handle in the for normal growth can be prepared as laboratory. follows: The best location for the cage is out of all drafts and direct sunlight. Rats Whole wheat flour 21 parts Yellow corn meal 21 parts should receive their sunlight only in the form of diffused radiation. The air must Whole oat flour 20 parts 20 parts be kept fresh and a temperature of 68° Whole milk powder to 72° is adequate. Sudden changes in Linseed Oil meal 10 parts temperature are harmful and should be Casein 3 parts avoided as much as possible. Some ar¬ Yeast 3 parts Calcium carbonate 1 part rangements must be made for keeping the animals warm in the school room Sodium chloride 1 part at night and on weekends during the This food is a dry powder and must be winter months. thoroughly mixed. Any reserve supply Each rat cage should be equipped with of food should be stored in a cool place. a food cup, water fountain, and, if feed¬ It is possible to supplement the diet ing or breeding records are to be kept, with carrots, lettuce, and other leafy a data card holder. The food cup can vegetables. be a small metal or glass cup if the The addition of vitamin E (wheat food is a dry powder type. However, germ, either dry or the oil) to the diet if the biscuit type of food is used, a of female rats used for breeding pur¬ small shallow wire-mesh basket can be poses is very beneficial in producing attached to one side of the cage. healthy litters of young. Water can be supplied in either a Laboratory rats frequently handled, small glass receptacle or a glass water become quite gentle, especially if they fountain. Such a fountain can be con¬ are cage-bred animals. Should it be structed in a few minutes by using either necessary to observe the animal, grasp it a test tube or a small round bottle, a firmly, but not quickly, by the middle
TURTOX Service Department Copyright, 1959, by GENERAL BIOLOGICAL SUPPLY HOUSE (IN'CO HP ORATED) 8200 South Hoyne Avenue Chicago 20, Illinois THE SION OF THE TURTOX PLEDGES ABSOLUTE SATISFACTION
Printed in U.S.A. GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
Young White Rats
of the tail and allow the animal to rest the young may be weaned. They will its feet upon the palm of the other hand. have eaten some food prior to this time so Another method of holding is to place that weaning is not too severe. The same the index finger under the neck, just in stock diet as that given the mother will front of the fore limbs and the other be relished by the youngsters. The young fingers about the belly between the fore rats may be left together until they are and hind limbs—the thumb around the eight weeks old, when the sexes should back of the neck. Always refrain from be separated to prevent breeding at this jerky or rapid movements when handling young age. animals, otherwise they may become Anyone working with young animals frightened. will need to know how to distinguish Use only healthy young adult rats for between the sexes at an early age. The breeding purposes. The female matures most reliable criterion is the distance sexually at about eighty days, but it is between the anus and the genital papilla; best to begin breeding when she is one it is greater in the male than in the fe¬ hundred days old. The litters will be male of equal age. In mature specimens larger and the young will be of better sex differences are easily recognized. size. The oestrous cycle of the rat is ap¬ The two sexes start out at much the proximately four days, subject to varia¬ same weight at birth. There is a discern¬ tion up to seven or eight days. The gesta¬ ible difference at forty days and after tion period is twenty-one to twenty-two that the male gradually increases in days, also subject to some variation. weight over the female. Females will usually mate immediately Rats normally live to be about three after casting a litter, if the male is pres¬ years old. Their most reproductive pe¬ ent. It is best to remove the male until riod is from three months to the end of the young are weaned before the females the first year. Commercially it is unwise are bred again. to keep and breed the worn out stock. Provide the pregnant female rat with Such animals should be elminated from an abundant lot of shredded newspaper the breeding colony when they have with which to build her nest. After the passed their prime. young are born handle the mother rat very gently. Any major disturbance may result in her eating the young. Mice: The young are quite immature at birth ; The albino mouse also has been used their eyes are completely closed and re¬ extensively in anatomical and physiolo¬ main so for fifteen days. At the time gical experimental work. A great deal of the eyes open, one can detect size differ¬ research on cancer has been done with ences and any runts should be discarded. white mice. Besides the white there are When twenty-eight to thirty-five days old other colored varieties such as chocolate
(40-2) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
220A528
Guinea-pigs make excellent laboratory 220A431 subjects for experiments in heredity and for testing the potency of vaccines, se¬ and black which can be used in cross rums and germ cultures. They are espe¬ breeding experiments, with the white cially recommended in elementary stu¬ mouse, to demonstrate inheritance of dies on reproduction because their care color. is so simple and they may be handled by Small wire cages such as recommended the students with little fear of being for rats can be used for mice. If wooden bitten or scratched. boxes are used, one-half inch of dry saw¬ Cages similar to those used for rats, dust should be placed om the bottom. All but larger, are required. A cage twenty- cages should be provided with dry hay four by sixteen by twelve inches with or shredded paper. The cages should sides and top of three-eighths inch wire be k*pt dry at all times. Keep the ani¬ mesh, and bottom of one-half inch wire mals at average room temperature and mesh is satisfactory. A door should be out of all drafts. constructed either on top or on one side Mice thrive on all sorts of dry grains, in order to remove the animal easily. corn, oats and wheat, also apples, lettuce The eages should be placed in a shallow and other green vegetables. They will metal pan to catch tne waste materials eat meat, but that should be avoided This pan should be cleaned frequently. because they may eat their young. Keep the animals at average room tem¬ The following mixture may be fed to perature about 72° F and out of all both rats and mice: drafts. Colds are usually due to sudden 20 parts cracked oats temperature and humidity changes as 10 parts buckwheat well as unclean quarters. 5 parts cracked corn Guinea-pigs are herbivorous in feeding 5 parts whole wheat habits and will eat a wide variety or 2 '/2 parts sunflower seed grains, nuts and greens. A satisfactory 1 '/2 parts millet diet is one composed of a mixt«re of 1 part peanuts grains, such as corn, oats and wheat with Water should be available at all times; succulent vegetables such as carrots, the glass water fountain is best since apples, lettuce, sweet potatoes, fresh there is less chance of spilling. grass and alfalfa hay. A mixture consist¬ Specimens should be sixty days old ing of equal parts of oats, wheat and before they are bred. The sexes may be barley, and a sufficient quantity of soy distinguished by the distance between bean or linseed meal, to form ten per¬ the anus and the genital papilla; it Is cent of the total, is a satisfactory food. greater in the male than in the female. When vegetables and fruits are plenti¬ The gestation period is twenty to twenty- ful the animals do not require water. two days. The males should be removed However, it is best to provide each cage from the cage when the young are born. with a non-spillable water dish. The young should be weaned when Allow the adults to become fully ma¬ twenty-one to twenty-eight days old. ture before breeding so that better lit¬ ters will be produced. They should not Guinea-Pigs: be bred before they are nine months old. Guinea-pigs are rodents native to The male may be allowed to remain in South America, but they have enjoyed a the cage with the pregnant female if world-wide distribution due to their great there is ample room, but if other cages desirability as laboratory specimens. are available it is better to separate They are docile animals, prolific and easi¬ them. The gestation period is sixty-five ly cared for. to seventy days. The young are well
(.40-3) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS developed at birth and can shift for hardy creatures. The animals sometimes themselves at an early age. They begin have external parasites which can be to eat solid food when quite young, and detected by the guinea-pigs’ scratching. tender leafy vegetables should be on The animals should be dusted with hand a few days after birth. They should pryethrum powder and the cages thor¬ be weaned when four weeks old. oughly cleaned and sterilized. Guinea-pigs are ordinarily healthy and
Please note: Postal regulations prohibit the shipment of live animals such as rats and mice by mail. Therefore, these animals can be delivered only via railway express. New express regulations set a minimum rate of $4.50 for any live animal shipment, however small. This rate is not excessive if six or more animals are ordered, but the cost of shipping one animal will be at least $4.50. 1SV72 Mice, White. Mature animals of pure strain but not pedigreed. Suitable for breeding or feeding experiments. Pair (male and female) $4.75
1SV74 Rats, White, Healthy, mature animals of pure strain but not pedi¬ greed. Suitable for average breeding or feeding experiments. Dozen.. .23.00 Pair 4.50
ISVTJfZ Adult Hooded Rat. A select strain of pure line “hooded” rats. De¬ sirable in introductory heredity ex¬ 220A525 periments—although they are not pedi¬ 220A525 Turtox Activity Cage. A rec¬ greed. Easily reared and serve wher¬ tangular, all-metal cage with a large ever rats are indicated in biology ex¬ revolving cylinder. This is a well-con¬ periment. Each (specify sex) 3.75 structed, roomy cage of our own man¬ Per pair (male and female) 6.50 ufacture. The wire mesh will retain mice or hamsters and the cage is large enough to accommodate white 220A431 Turtox Rat Cage. Cage is rats and small squirrels. Size 16 in¬ round, 9 inches high by 9 inches in ches long, 10'4 inches high and 9 in¬ ches wide. Complete with removable diameter. Made of heavy galvanized metal try and sanitary water fountain. mesh with removable bottom pan. Each $19.95 Each 10.95 55V80 Rat Grozc'iny Ration. A high Per unit of two cages 21.00 grade ration for feeding growing rats or mice. Contains all the necessary 220A528 Turtox Economy Cage for mice proteins, carbohydrates, fats, minerals and hamsters. Easy to clean. Size and vitamins. We recommend this as a 8%" x 6" with a 4)4" opening. diet for growing animals. Mixed and ready for use. Per lb 1.00 Each 2.25 Per 5-lb. package 4.25 55V32 Turtox Guinea Pig and Rabbit 220Alp Animal Cage Breeding. A cage Food. A prepared food containing the suitable for breeding rats or guinea essential nutrients for growth and re¬ pigs or for group feeding experiments. production of guinea pigs. By using Size 18 x 12 inches by 9 inches high. this food, greens need be fed the ani¬ mals only once or twice a week. Lab¬ Made of % inch mesh galvanized iron oratory tested. Directions for feeding with bottom of inch mesh, resting in supplied with each shipment. detachable galvanized iron pan. Top is hinged. Card holder for 3 x 5 data card. 1-pound package 60 5-pound bag 2!50 25-pound bag 9.50
All prices are f.o.b. our laboratories and are subject to change without notice.
(40-4) TURTOX SERVICE LEAFLET No. 20
NOTES ON MARINE AQUARIA
The biology student living on the sea- thoroughly cleaned and made ready be¬ coast has a great advantage over his fore the shipment of sea-water and living fellow in an inland laboratory, for there specimens arrives. The tanks should be can be no comparison made between the located in a place where a fairly low interest-arousing qualities of a star¬ temperature can be maintained and where fish dripping with and smelling of they will receive little or no direct sun¬ formalin and those of a living starfish light. crawling over the submerged rocks in a It is a good idea to divide the shipment clear tide-pool. There is no reason, of sea-water and living specimens be¬ however, why students living far from the tween two tanks. This allows one to de¬ seacoast should not have the opportunity termine which animals will live together of seeing living examples of some of the in harmony and, should any forms be smaller marine animals. Salt-water injured, they may be kept in a separate aquaria are now used in hundreds of aquarium until they recover. inland schools and the living marine animals can be shipped successfully at any time during the colder months of Sand the year. If you use sand other than that secured from an ocean beach, be sure that it is a pure silica sand and wash it thoroughly Collecting and Shipping and repeatedly to remove any mud or Teachers who are fortunate enough to organic matter before it is placed in your live near the seashore are usually able to aquarium. collect an interesting variety of living marine animals at any season of the year, Sea-water and, if the school laboratory is located nearby, the transportation of the speci¬ Natural salt-water from the ocean is mens presents no problems. Wooden or supplied with our aquarium sets and we enameled pails are best for use in marine do not advise the use of synthetic sea¬ collecting, and, of course, glass jars of water unless large quantities are re¬ various sizes are suitable for small forms. quired. However, some teachers will find (Do not use metal containers.) Collect it economical to prepare synthetic sea¬ small-sized specimens and do not crowd water and, in such instances, the following too many into a small amount of sea¬ formula may be used: water. It is far better to return to the Distilled (or rain) water 10 gal. laboratory with one small starfish alive Sodium chloride, C. P 45V2 oz. than with a pail-full of dead or dying Potassium chloride 1)4 oz. specimens. Calcium chloride 2 oz. Magnesium chloride (dry) 8% oz. Magnesium sulphate 11% oz. The Aquarium Tank Bicarbonate of soda 1/5 oz. The best container for a small marine After thoroughly mixing the above, aquarium is a rectangular all-glass tank, add: although the standard metal-framed tanks Potassium nitrate 1/5 oz. with slate bottoms and glass sides may be Sodium phosphate 10 grains used provided they are so constructed Iron chloride 5 grains that no water comes into direct contact Natural sea-water 1 gal. with any metal parts. The best size for The reason for adding the natural sea¬ a beginner is a tank of from six- to ten- water is not entirely clear; but it is un¬ gallon capacity. After one has had some questionably necessary. Some vital ele¬ experience with a small aquarium, the ments are apparently lacking in the larger sizes may be used. manufactured sea-water and the addition The tank or tanks to be used should be of the natural water remedies this. It
TURTOX Service Department Copyright, 1959, by GENERAL BIOLOGICAL SUPPLY HOUSE (Incorporated) 8200 South Hoyne Avenue Chicago 20, Illinois
THE SIGN OF THE TURTOX PLEDGES ABSOLUTE SATISFACTION
Printed In U.S.A. GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS has been suggested that natural sea-water Aeration of the Water contains a substance corresponding roughly to the vitamins in foods. The first and most important rule to After the synthetic sea-water has been follow in planning salt-water aquaria is made up, it should be placed in tightly to remember that most marine animals corked glass carboys and kept in a dark require more oxygen than do most fresh¬ place until wanted for use. water forms. The main reason for this is that the Maintaining the Right Concentration marine forms used in aquaria are usually When the tank is filled with sea-water, collected in tide-pools along surf-swept the level of the water should be marked beaches where the oxygen content of the on the outside of the glass by drawing a water is unusually high. Therefore, fewer line to coincide with the surface of the inhabitants should be placed in each tank water. As evaporation takes place, pure than would be the case if one were deal¬ distilled water should be added to bring ing witli fresh-water animals. The tend¬ the water up to the original level. In ency is always to overcrowd an aquarium large tanks, the water should be tested and, although this may cause eventual from time to time to see if distilled water failure of a fresh-water tank, it is quickly is needed to replace the loss by evapora¬ and completely fatal in a salt-water tion. Natural sea-water should show a aquarium. reading of 1.025 when tested with a hydrometer. Any needed adjustment Large public exhibition aquaria are should be made weekly or oftener. usually planned so that there is a large reservoir of salt-water furnishing enough water for a continuous, though rather Temperature of the Water slow, flow through the exhibition tanks. The proper temperature control is of Compressed air vents are usually placed utmost importance in the success of a so that the water is aerated thoroughly marine aquarium. In general, the water while in the main storage reservoir. How¬ temperature should be maintained at 55° ever, the size and cost of such a system to 60° F and a temperature ten degrees as this renders it impossible for the lower than that is often better. average school laboratory. Sudden changes of temperature are usually fatal to aquatic animals. If your Several small and inexpensive aerating shipment of living marine animals ar¬ pumps are now available and one of these rives during the winter when the weather will be of great help in aerating the is cold, the temperature of the sea water water in your aquarium, especially dur¬ in their container will probably be very ing the first week or so until the animals little above 32° F. This should be raised become “acclimated” to their new sur¬ very gradually and, in no case, should the roundings. (Turtox will furnish informa¬ animals be transferred suddenly to much tion and prices on suitable aerators for warmer water. A good plan is to allow any size tank.) a period of at least 24 hours in which to allow the temperature of the water to A marine aquarium may, of course, be rise gradually. “balanced” just like a fresh-water aqua¬ rium if the oxygen given off by the plants Many suggestions have been offered in is sufficient to supply the needs of the regard to the matter of keeping the school animals. The green alga, Ulva, commonly marine aquarium water cool; but the one called sea-lettuce, is helpful as an oxygen considered most feasible is that in which producer, as is also Cladophora. Marine the aquarium is placed on the window diatoms are particularly desirable and we sill where it will not receive direct sun¬ now include them in the sand supplied light. The window is then opened until with our larger sets. In general, however, about two or three inches of the glass the “balancing” of a marine aquarium side of the aquarium are exposed to the comes about gradually and, at the start, it out-of-door conditions. The open space is highly desirable to aerate the water by on either side of the tank is then blocked artificial means. off with wooden panels and strips of felt so that the room temperature will not be affected. The advantage of such a system Light as this is that it is possible to maintain a lower degree of temperature in the Salt-water aquaria require much less aquarium than could otherwise be at¬ light than do fresh-water aquaria and tained. they should receive very little direct Tropical forms must, of course, be sunlight. Except when it is desirable to provided for according to the conditions watch the inhabitants, the aquarium found in the water where they are should be shielded by cardboard on three collected. sides to keep out very strong light.
(20-2) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
A small marine aquarium containing starfish, sea urchins, sea anemones, Ulva and Fucus.
—silicon sponges ; Coelenterata—Obelja, The Animals Clava, Sea anemones, Corals ; Platyhel- With the aquarium tank made ready minthes—Small Turbellarians found on and in its permanent location, the pre¬ Ulva; Nemathelminthes—free living; Ro- liminary preparations are completed and tifera; Bryozoa; Annelida—Serpulids or the living marine animals may be or¬ tube worms, Nereis, Climenella; Mol- dered. Under the ideal conditions exist¬ lusca—Snails, Limpets, the nudibranch ing in very large aquaria, almost any Aeolis, Clams (both Mytilus and burrow¬ marine forms will live; but the marine- ing forms), Pecten, Chitons; Arthropoda aquarium enthusiast who is experiment¬ —Barnacles and many small Crustacea; ing with small tanks should attempt to Chordata—Ascidians; all coming from secure the more hardy animals which will pool or low tide areas.” live for a while, at least, under somewhat adverse conditions. Among the best Feeding the Animals small-aquarium inhabitants are small marine snails and barnacles. Starfish, To feed the animals, it is sometimes sea urchins, sea cucumbers and sea ane¬ best to remove them to a separate dish mones will usually live for a few weeks filled with salt water of the same tem¬ and under carefully controlled condi¬ perature as that of the aquarium. Wood¬ tions may be kept in aquaria for much en forceps are best for handling the ani¬ longer periods. Small crabs will often mals. Twice a week is often enough to thrive where more exacting forms die, feed the various forms. Small pieces of and Obelia will often live and reproduce macerated oysters, clams or fish make a new colonies in very small aquaria. fine food and they should be dropped near the mouth of the animal by means Dr. Lyell J. Thomas, who has done of forceps. The juice of oysters and some very interesting work with marine clams also makes a fine food and can be aquaria at the Zoological Laboratory of dropped by means of a pipette into the The University of Illinois, lists the fol¬ mouths of such animals as the Metridi- lowing animals which have been found to um, Thyone and Cucumaria. Fresh water do well in small marine aquaria under clams may be used as well as salt water laboratory conditions: clams and even small pieces of fresh “Protozoa—a long list have been identi¬ water fish will be readily devoured. As fied and offer the inland protozoologist a soon as the animals have been fed, they new and fertile field for study; Porifera may be returned to the aquarium and
(20-3) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS the glass plate put back on top of the of them; (2) Keep the water temperature tank. low; (3) Aerate the water constantly If you wish to feed the animals with¬ and (4) Feed sparingly. The Turtox out removing them from the main tank Service Department will gladly answer (and this is advisable, of course, in many your questions and offer help in the es¬ cases), put in very small amounts of tablishing and caring for marine aquaria food and remove promptly any that is of any size. Write to us if there are any not eaten, or otherwise it will decompose points on which you wish additional in¬ and quickly foul the aquarium. formation. Temporary Marine Aquaria Literature “Culture Methods for Invertebrate Even though it may require consider¬ Animals.” Published by Comstock Pub¬ able effort to maintain salt-water aquaria lishing Company. for long periods of time in a small lab¬ “The Aquarium Book,” by E. G. oratory, the teacher will find that a few Boulinger. Published by D. Appleton attempts along this line are well worth & Co. while. The tanks containing sea-water “Goldfishes and Tropical Fishes,” by can be maintained throughout the school W. T. Innés, published by Innés and Sons. year, and from time to time, assortments Philadelphia. of a few living marine forms can be ordered. Even though some animals are “Fishes in the Home,” by Mellen, pub¬ short-lived, the students will have had lished by New York Zoological Society, an opportunity to study living starfish, New York City. sea urchins, anemones and other marine “Guide to the New York Aquarium,” forms which they might otherwise never published by New York Zoological So¬ have seen. The result of even a few hours ciety, New York City. spent in this way will be a real interest “Living Marine Animals for our In that preserved specimens could never land Laboratories.” A report by Lyel) have awakened. J. Thomas, appearing in the January, 1925, issue of Transactions of American Microscopical Society. Summary “Care of Small Salt-water Aquaria,” The really essential points to keep in by I. M. Mellen. Published by New York mind are few; but they are very impor¬ Zoological Society, New York Aquarium, tant: (1) Use small specimens and few New York City.
For information on shipments of living marine specimens write to: Coral Reef Exhibits, P.O. Box 23, Coco¬ Miami Tropical Aquarium, 982 South¬ nut Grove 33, Florida west 3rd St., Miami 36, Florida Greater Miami Fisheries, 3475 N.W. Supply Department, Marine Biological 187th St., Opa-Locka, Florida Laboratory, Woods Hole, Massachusetts. Global Aquarium, 65 Mt. Vernon St., Ridgefield Park, N. J.
205A20 Turtox Special Aquarium. 205A6 Aquaditioner Air Pump. A Size 18" long, 10%" wide and 9%" good low-price aerator for tanks up high. Capacity, 6 gallons. Metal frame with sides, ends and bottom of heavy to 50 gallons. Operates on 110 to 120 glass $15.95 volts, A.C. or D.C. $6.50
205A 201 Turtox Special Aquarium. 205A601 White Mist Aerator. An ex¬ Same as Number 205A20, but larger, of 10 gallon capacity 19.95 cellent aerator for use on 110 volts, A.C. Complete with 5 feet of rubber 205AU0 Aquarium Forceps. Wood, tubing and one air breaker 21.00 18%" long. Each 2.25 205A578 Temperature Control Unit. 205A 6 6 Aquarium Cement. Pound A combination unit, consisting of 75 can 95 watt heater, thermostat and indicator light. For 110-120 volts, A.C. All prices are f.o.b. our laboratories and only 9.50 are subject to change without notice.
(20-4) TURTOX SERVICE LEAFLET No. 5
STARTING AND MAINTAINING A FRESH-WATER AQUARIUM
Photograph of the No. JtSVll Turtox Aquarium Outfit, as listed on page four of this leaflet.
Starting and maintaining a fresh-water aquarium in the school laboratory is easily accomplished by anyone who is willing to devote a little thought and care to the work. The pleasure and practical teaching advantages to be derived from one or more aquaria containing interesting plant and animal specimens is well worth the time and effort expended. Before starting your aquarium, however, make sure that you have the correct type of tank, the right quantities of aquatic plants and animals. Keep in mind the basic requirements of the specimens to be included in such a demonstration, because a fresh-water aquarium is a biological association of plant and animal life. The following brief outline states the procedure to follow in establishing a fresh¬ water aquarium. If these directions are followed closely, no particular difficulty should be encountered.
TURTOX Service Department TURTOTMIUCTS Copyright, 1959, by GENERAL BIOLOGICAL SUPPLY HOUSE (INCORPORATED) 8200 South Hoyne Avenue Chicago 20, Illinois THE SIGN OF THE TURTOX PLEDGES ABSOLUTE SATISFACTION
Printed In U.S.A. GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
Equipment. The only absolutely essen¬ tial piece of equipment is a suitable aquarium tank; a rectangular tank with metal frame, glass ends and sides, of about 6 to 9 gallon capacity is recom¬ mended. Small, round aquaria are not satisfactory (visibility is poor through curved glass), and the round (ish globes are useless. In addition to a suitable tank, the following items will prove their usefulness: dip-net for trans¬ ferring fish, one pair of long aquarium forceps, one pair of plant snips and a syphon tube.
HOW TO START THE FRESH-WATER AQUARIUM 1. Clean the tank thoroughly, remov ing all dust, grease, etc., from the glass. If the tank has previously been used as an aquarium wash very thoroughly with soap and ammonia water. Rinse with clear water three or four times. 2. Clean thoroughly, in running water, sufficient aquarium sand or gravel to cover the bottom of the tant to a depth of one to two inches. Con tinue washing until all debris and sol¬ uble matter has been removed. Place the sand in the aquarium. 3. Now add the water. Use clear pond water if possible; if this cannot be obtained, use tap water which has stood in open containers for a day or so. Pour the water into the aquarium (using a sheet of paper as shown in illustration) until it is 8 or 8 inches deep. After it has been planted add more water to fill the aquarium. (Im¬ portant: Chlorinated city water is not safe to use unless it is allowed to stand in open containers for 48 hours.) 4. The aquarium should be placed where it will be exposed to strong dif¬ fused light. Direct sunlight for an hour or so each day is usually not harmful, but neither is it necessary. North or East exposures are usually best. 5. The aquarium is now ready for planting. Plants are used for a variety of functions in an aquarium. They pro¬ vide a natural habitat and protection for small fishes. Plants utilize some of the excreta thrown off by the fish and other animals while they, in turn, are used as food by some species of fish and snails. They also promote the growth of microorganisms and other small animals.1 It is well to use plants rather spar¬ ingly at first, remembering that they will soon grow and spread. If too many are crowded in at the beginning some
(S-2) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
may die and decay, thereby fouling the water. All plants will not thrive in an 5. Obtain dome rooting planta from aquarium and it is best to secure tank- Turtox grown (not wild) aquatic plants of kinds that are known to grow well in aquaria. Vallisneria, Sagittaria, Elodea (Anacharis), Myriophyllum and Ca- bomba are all good. Many other plants can be grown successfully in an aquari¬ um, but the ones mentioned give variety and are dependable. 6. Plant the aquarium as follows: Take several of the rooted plants (such as Vallisneria and Sagittaria), spread the roots out on the sand and cover tlwoeria oagitUria Ludtfigia them up to the crowns, pressing them down to secure anchorage. Add several 6. Also dome non-rooting planta stalks of the non-rooted plants (Elodea, Myriophyllum and Cabomba), weighting the lower ends down with small stones or with pieces of lead. Arrange the upper parts of the plants so that they float freely in the water, and remove all dead or broken leaves. Now fill the aquarium with water to within one inch of the top. If the conditions are right, growth should be noticeable in about two weeks after planting. If the water does not become clear in a day or so it is usually an indication that it contains some dead Anacharis Cabomba Myriophyllum and decaying plants or other organic matter. 'y, Place tank in permanent location. Partly fill \dith clear Water, set plants in grav'd 7. Allow the aquarium to stand for a few days until the water has cleared, when it will be ready for its animal population. (Snails, fishes, etc., can be placed in the aquarium immediately after planting if necessary, but it is best to wait until the water is clear.) 8. In stocking an aquarium several things must be borne in mind. First, use only animals which get along to¬ gether; predaceous forms must be kept by themselves. Second, do not use ani¬ mals (or plants) which naturally live only in running water, as they will not Ô Complete filling of tank with water and cover live in the close confinement of an wtthglass plate. 'Add no animals until water aquarium. Third, do not overcrowd. is clear and the plants are growing. Fourth, do not use animals which will stir up the sand on the bottom and keep the water cloudy. 9. Animals. Fish. It is important not to over-crowd an aquarium with fish. A good rule to follow is one inch of fish to one gallon of water. The average six-gallon tank will support six one-inch fish or three two-inch fish. This rule cannot be followed in all cases. The number of fish used must be governed by the number of other animals to be included in the GDIERAL BIOLOGICAL SUPPLY HOUSE-7 6HÜ.AaT Ç^lilAcE-CHIC Ago.
(5-3) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS aquarium and the surface area of the 10. Watch the aquarium carefully for water. It is usually advisable to use the first few weeks. Remove at once any several small fish rather than one large dead animals and clip off all dead parts one. Small native fishes (minnows, dace, of plants. After the plants have become bullheads and sunfish) are far more in¬ established it may be necessary to remove teresting than the sluggish goldfish. If fish some of them to prevent over-crowding. are not obtainable, snails, newts and aquatic insects will add much life and FEEDING THE AQUARIUM interest to the aquarium. ANIMALS Snails. Six to a dozen snails may be kept in a 6-gallon aquarium. Snails are This subject is discussed in Turtox valuable as scavengers and will help keep Service Leaflet No. 23 and will be men¬ the algae from the glass. Various species tioned only briefly here. The most im¬ of pond snails (egg layers), and red portant rule to be followed in caring for snails, may be used. If possible, one or aquarium animals is—Feed Sparingly. two viviparous (liv<" bearing) snails Because higher animals must be fed often should be included. it is but natural that one should view with alarm the prolonged fasts of the Clams. One small clam, an inch to two cold-blooded animals. However, these inches long, may be used. This is an forms can go for long intervals without interesting form to study, for the inhalant food and remain perfectly healthy. Most and exhalant siphons show perfectly. The of the small native fishes will do well on clam should be watched closely to see a balanced food, such as the Turtox that it is living, as it will disintegrate Natural Fish Food. They also relish quickly and foul the aquarium if it should living crustaceans, such as Daplmia or die. Newts. Several small newts can be kept Cyclops, and finely chopped live earth¬ in the aquarium. They are active, inter¬ worms. Newts will eat tiny pieces of raw esting and relish small bits of raw meat meat, raw fish or small pieces of living as food. earthworms. The snails and clams neeii Reference: not be fed, as they can shift for them¬ 1 Atz, J. W. "The Functions of Plants in Aqua¬ selves in an aquarium containing plenty ria", the Aquarium Journal, Vol. of living plants. They also feed upon 21, No. 2 and 3, February and March, 1960, pp. 40-43; 66-60. the excess fish food.
4SV10 Turtox Aquarium Set. A thor¬ 205A41 Aquarium Forceps, Wooden oughly dependable collection of forceps, 18% inches long. Each plants and animals for the balanced $1.10 fresh-water aquarium. Includes suf¬ ficient material to stock a standard 205A54 Aquarium Dip Net, Each 6 to 9-gallon aquarium tank. 75 Plants: Animals: 205A47 Aquarium Siphon. Flow of Sagittaria Snails of two kinds water starts automatically. Made of Ludwigia Two aquatic Newts glass with rubber connections. Vallisneria One small clam Length, 12 inches. Each 1.40 Cabomba (Or other aquatic Myriophyllum animal.) Write for current information on Water Poppy tanks, aerating pumps, air breakers, (Or other heaters and thermostats. plants may be substituted.) FOODS AND REMEDIES Set as described $4.75 45V11 Turtox Aquarium Outfit. In¬ 55V21 Turtox Natural Fish Food. A cludes the special new Turtox Aqua¬ balanced food. Per 100 cc jar .75 rium of 6-gallon capacity made of 55V24 Shredded Shrimp. Per 100 CC. polished bulb-edged glass with slate jar 50 bottom, one-piece metal frame and removable glass top (see page 1), 1 55V41 Fungus Cure for Fishes. Per bag Turtox washed aquarium gravel, ounce of concentrated crystals. .50 1 jar Turtox Natural Fish Food, and 1 aquarium dip net. Complete outfit 55V42 Salt Bath for Fishes. A com¬ with instruction leaflet .. 18.50 bination of three mineral salts. Per ounce 50 45 VI2 Complete Aquarium Outfit with Plants and Animals. Consists 55V44 Medicated Aquarium Hi-Ball. of Nos. 45V10 and 45V11 de¬ For neutralizing acidity in the scribed above. Both for 23.00 water. Each 25 All prices are f.o.b our laboratories and are subject to change without notice. (5-4) TURTOX SERVICE LEAFLET No. 42
LABORATORY DISSECTIONS To the scientist, the word “dissection” 3. The Right Frame of Mind. Every has a meaning far different from the teacher of Zoology with a class of stu¬ idea that it conveys to the beginning dents taking the subject for the first student in Biology. The word literally time finds certain members of either sex means “to cut apart,” with the word ‘'cut” who have a natural or feigned aversion given as a synonym. However, when one for handling laboratory specimens. Such learns that synonyms for “cut” include students usually receive little sympathy such an array as “carve, chop, cleave, from the average teacher, for he knows gash, hack, hew, sever, shear, slice, sunder that interest in the specimen and in the and whittle” as well as “dissect,” it is subject soon submerges any aversion on obvious that the instructor does not have the part of the student. The student all of these in mind when he calls for a himself should proceed with his dissec¬ careful dissection of some laboratory tion with the exploring spirit—here is form. The word anatomize expresses something new of which he has a limited more clearly what the scientist has in knowledge and he should be determined mind when he speaks of a dissection, for to learn by himself the structure, mech¬ this implies the separation of organs anism and functions of the specimen be¬ and tissues in such a manner as to give fore him. a true understanding of the various parts of the specimen. Prerequisites of a Good Dissection 1. Good Materials. The first pre¬ requisite of a good dissection is good material. Specimens that are imperfectly preserved, with organs and tissues in various stages of disintegration, obviously are unfit for dissecting purposes. It is also desirable that the specimens be properly straightened for convenience in pinning out or fastening in the position in which they will be dissected. For ex¬ ample, an earthworm that is not straightened and relaxed before preserv¬ ing, or a frog that is badly distorted, makes dissection just that much more difficult. 2. Proper Instruments. By proper instruments it is not meant that they must be expensive. It does mean that one should have the necessary instruments for certain purposes. For ordinary dis¬ sections called for in elementary Zoology courses one pair of fairly fine-pointed Fig. 1. Diagram of proper method of pinning scissors, one scalpel, one forceps, two an earthworm for dissection. needles and one probe are sufficient. Where iiigher vertebrates are dissected Dissecting the Specimen various types of the above instruments In practically all cases the specimen are desirable, with the possible addition should be fastened to the wax-bottomed of bone forceps and other special pur¬ dissecting tray by means of pins. The pose instruments. The purpose of the in¬ pins should be inserted obliquely as strument and the use to which it is to shown in Figure 1. There are three be put is most important, although the reasons for this—First, when inserted quality should be good. For student use obliquely the pins are braced against any dependable instruments may be procured tension set up in the specimen and will at a reasonable price. not easily be pulled out. Second, the
TURTOX Service Department Copyright, 1959, by GENERAL BIOLOGICAL SUPPLY HOUSE (Incorporated) 8200 South Hoyne Avenue Chicago 20, Illinois THE SION OF THE TURTOX PLEDGES ABSOLUTE SATISFACTION Printed in U.S.A. GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
specimen does not have a tendency to with a slight spreading motion in the di¬ "creep” up the pins and change position. rection taken by the parts being dis¬ Third, the pins are out of the way of the sected. The needles may be used at times hands and instruments, thus permitting in the same way, depending on the free access for working on the specimen. delicateness of the structures involved. Opening the Specimen. After the speci¬ Use the scalpel or scissors only when ab¬ men is securely pinned the first step solutely necessary. Avoid pulling, pinch usually is to open the body cavity. At ing and picking at the specimen in an this time great care should be taken. aimless manner. Make every move with Always note carefully the laboratory di¬ a definite end in view. As O. W. Holmes rections for opening the specimen and has so aptly said: “Let the eye go before follow them explicitly. If scissors are the hand, and the mind before the eye.” used, carefully insert the point so that it does not penetrate deeply. Then hold¬ Orderliness and Cleanliness ing the ‘‘finger ring” of the scissors Conducive factors in good laboratory down low, thus keeping the point up work ai-e orderliness and cleanliness. against the inside of the body wall, care¬ Determine the most efficient arrangement fully extend the cut to the point indicated of all materials and instruments used in the directions. The thumb and middle and keep them in good working order. finger should be used in the scissor rings, Cleanliness of instruments and hands and with the index finger serving to steady the disposal of dissected parts, or speci¬ the instrument in cutting. Certain small mens no longer needed, help to keep one specimens, as the earthworm, should be in the proper frame of mind for good pinned out progressively as the cut is work. Slovenliness in the care of ma¬ made. terials and instruments is only too likely Further Dissection. Once the body to be a reflection of the student’s state wall has been cut through and the speci¬ of mind. men properly pinned, the greater part of It is not amiss at this point to remind Ihe dissection should be carried on with the student that the essential feature of non-cutting instruments, thus avoiding laboratory work is to learn to observe the destruction of important structures correctly, to remember what he observes by cutting. Once parts are severed, and with his assimilation of facts to com¬ especially blood vessels and nerves, their pare the various specimens that he correct relationships are likely to be lost. studies in the laboratory. For these Instead of cutting, use the closed points processes there is no time when he can of the forceps for pressing one way study and retain facts with greater against one of the parts to be separated facility than while he is actually making and the probe on the other part for ef¬ the dissection. The student who follows fecting the separation. In this way the his laboratory directions intelligently connective tissues that bind the parts to¬ and who faithfully executes those direc¬ gether can be loosened and the structures tions, studying and understanding as he clearly exposed. In separating muscles, goes, is the one who will get the most blood vessels and nerves pull the probe out of his laboratory work.
TURTOX ELEMENTARY DISSECTING SET. 308AS01 Elementary Dissecting Set. Our most popular dissecting set, many thousands of winch are sold each year. The instruments and case are of dependable quality, and this set has proved very satisfactory for beginning zoology and biology classes, in both high schools and colleges. The following instruments are in¬ cluded: One pair scissors, one pair forceps, two teasing needles, one all-steel scalpel, one 6-inch English and Metric ruler and one high quality waterproof case. Price each $3.95 310A5310 Dissecting Pan. Made of non-rusting galvanized sheet metal with J4 inch wax liner for pin¬ ning. Complete with rings to which specimens can be fastened. Size, 7 by 11 x 1J4 inches. Each 2.75 Dozen 31.25 310A56 Dissecting Pins. Extra heavy, 1)4 inches long Per % lb. box (approximately 600) 1.85 For a complete listing of dissecting instruments, refer to your Turtox Catalog. Drawings of the anatomy and skeletons of all of the laboratory animals are now available. Ask for the Three-Way Checklist of Turtox charts and biological drawings. All prices are f.o.b. our laboratories and are subject to change without notice.
(42-2) TURTOX SERVICE LEAFLET No. 6
GROWING FRESH WATER ALGAE IN THE LABORATORY
Fresh-water algae are of very gen¬ luxuriant forms like Cladophora may eral distribution and are found in nearly soon overstock themselves and the ex¬ every type of damp and aquatic habitat. cess material must be removed from Ditches, ponds, rivers, lakes and marshes time to time. of any locality abound in a great variety Cultures may be started at any time of forms, both of the unicellular and of the year. In winter bring in some multicellular types, Some algae grow mud over which the desired form was on damp earth or rocks and some kinds growing the previous season. Sticks or make up the greenish covering which stones may also be used. Place it in a appears on the bark of trees. Most of jar and add tap water, distilled water, the more conspicuous forms are inde¬ or rain water as the case may require. pendent, either free floating or attached Cultures of some forms, such as Chara in tufts or mats to the substratum, but and Nitella, have been obtained from there are also epiphytic and endophytic mud collected several years before. Cul¬ species. Many of the smaller forms are tures started in this manner usually yield attached to the other water plants or are a variety of material. found in the loose sediment and debris If you have healthy culture, do not of the substratum. throw it out if the algae should disappear Many kinds of algae are easily cul¬ Seasons of dormancy occur in nature— tured in the laboratory. Some kinds will look in the bottom of the jar for spores. grow well if brought indoors and placed Let the water evaporate, cover the jar in containers of pond water or in a for protection, and set it aside. After a balanced aquarium; other field-collected period of a month or two add more algae are more difficult to culture and water and you will likely have your cul¬ can be maintained over long periods only tures again. Lengths of dormancy pe¬ by the use of nutrient solutions, and the riods vary. In Volvox it is quite long; pure-culture technique. For most kinds in Oscillatoria it is only a couple of of algae, large finger bowls or battery months; in Cladophora there may be no jars are good culture containers; some dormancy. Warm weather makes it dif¬ robust forms such as Spirogyra and ficult to keep some cultures of algae in Cladophora are best grown in aquarium good condition. Refrigeration, after col¬ tanks. lection, will make it possible to study the When possible, use the water in which material for a week or more instead of the algae were growing, since sudden just a day or two. changes in kinds of water are injurious. The method used in collecting algae is Where additional water is needed use different from that used for aquatic zo¬ distilled water, or let the tap water run ological specimens. Care must be taken for several minutes before filling the jar, to avoid metal containers, specially on since water standing in pipes, or other long, warm trips. Ample water must be metal containers for that matter, is used if the containers are to be sealed. harmful to most algae. Add new water However, forms like Spirogyra, Mou- gradually, to compensate evaporation. geotia, Zygnema, Cladophora, Hormiscia Do not put too much material into a and Vaucheria may be rolled in wet jar. Actively growing material will in newspapers or magazines and carried crease and gradually accommodate itself thus. to conditions. The excess in an over¬ abundant supply will be choked off and Blue-Green Algae the consequent decay will cause fermen¬ tation and general fouling of the culture, Oscillatoria is readily found in stag¬ ultimately killing the whole. Likewise, nant water, watering-troughs, damp
TURTOX Service Department UCTS Copyright, 1958, by GENERAL BIOLOGICAL SUPPLY HOUSE (INCORPORATED) 8200 South Hoyne Avenue Chicago 20, Illinois THE SIGN OF THE TURTOX PLEDGES ABSOLUTE SATISFACTION Printed in U.S.A. GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS earth, flower pots, and in many other water. Droppers (for transfers) may habitats. It is recognized by its dark be steamed and kept in a wide-mouth blue-green, or blackish color. Oscillatoria screw-cap jar. is one of the easiest of plants to keep Store cultures in a room where there in the laboratory. Place a little of the is a plentiful supply of daylight, at tem¬ material in a container partly filled with peratures not in excess of 80° F. (Op¬ water; cover with a lid to avoid unneces¬ timum 70°-75°F.) If, because of the sary contamination from dust. By add¬ season or for other reasons it Becomes ing water from time to time to compen¬ necessary to use artificial light, we sate for evaporation, the cultures should recommend the Sylvania 150W, 120V keep indefinitely. Spot Bulb (Turtox No. 375A97) at a Nostoc is frequently found in lakes distance of about eight feet from the and also occurs on damp earth. It is easy culture shelf. to maintain in laboratory cultures, and For those who wish to compound will keep in good condition for a month their own media, the following have or more if kept under refrigeration at been suggested in the literature, and we a temperature of about 40° F. recommend reading the references cited Rivularia is found attached to the at the end of this leaflet. leaves and stems of various aquatic Knop’s solution, for forms that prefer plants. It often occurs in laboratory an acid medium: aquaria. Magnesium sulphate, Oloeocapsa is found in gelatinous MgS04.7H20 0.25 g. masses, sometimes floating in ponds, but Potassium phosphate, more often as a coating on wet rocks. KH2P04 0.25 g. It is easily maintained in laboratory Potassium chloride, KC1 0.12 g. cultures. Calcium nitrate, Green Algae Ca(N02)2.4H20 1.00 g. Volvocales. Most of the members of To prepare, measure out one liter of this group and some of the related fami¬ distilled water and dispense about 250cc. lies are best grown in pure culture, into each of 4 flasks. Dissolve the chem¬ using nutrient solutions. The Turtox icals separately in these portions of wa¬ “Universal” Concentrate has been de¬ ter and combine into one volume—add¬ veloped for culturing many of these ing the calcium nitrate last. There should forms. Its use (described below) en¬ be no evidence of precipitation if the ables anyone to grow and maintain per¬ individual solutions are well mixed as manent pure-line cultures, including they are combined. To complete the me¬ such difficult forms as Volvox and Eu- dium, add one drop of freshly prepared dorina. (The Turtox strain of Volvox 1% ferric chloride solution. Experimen¬ aureas has been cultured continuously tation will best determine the most suit¬ on this medium from the spring of 1949 able concentration for any particular until now, 1958.) alga—which may vary from full strength Turtox 4-Unit “Universal” Concen¬ to a 1:10 dilution. The pH of this solu¬ trate 61V170 Set “A”. Makes one liter tion will be about 6.4. of medium for growing Volvox, Eudo- Knop’s solution (modified), pH 7.6. rina, Pandorina, Chlamydomonas, Spiro- Magnesium sulphate, gyra, Zygnema, etc.: The medium is MgS04.7H20 0.1 g. made from the concentrate by mixing Potassium phosphate, all of the Unit I with 925cc. of distilled K HP0 0.2 g. water. This is followed by the addition 2 4 Potassium nitrate, KN03 1.0 g. of Unit II and, after further mixing, Calcium nitrate, by Units III and IV. The culture fluid Ca(N02)2.4H20 0.1 g. is then ready for immediate use. Prepare as outlined above and add Turtox 4-Unit “Universal” Concen¬ one drop of 1% ferric chloride solution. trate 61V170 Set “B”. Makes five When combined with agar, the fol¬ liters of medium. Prepare in the same lowing formula is well suited for the way as Set “A”, but add the units propagation of Chlorella, Gloeocapsa, to 4625cc. of distilled water. Cultures Pleurococcus, Desmids, Diatoms, etc. may be put up in battery jars (with Ammonium nitrate, glass plate covers to prevent the en¬ NH4NO, 0.5 g. trance of gross contaminants) or in Potassium phosphate, small, stacking finger bowls—allowing KH2P04 0.2 g. about % inch of air space in each bowl, Magnesium sulphate, and covering the uppermost dish with MgS04.7H20 0.2 g. a glass plate. All glassware previously Calcium chloride, used for culturing of algae should first CaCl .2H 0 0.1 g. be washed and then steamed 30 min¬ 2 2 utes to avoid contamination. New glass¬ Agar 12.0 g. ware may be made ready for use by Distilled water 1000.0 cc. washing it with detergent and hot water, 1% ferric chloride solution 0.1 cc. followed by a rinse in weak hydrochloric (Note: the agar may be omitted if a acid solution and a final rinse in tap liquid medium is desired.) (6-2) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
Soil-Water Medium (a modification The formation of zoospores may be of the Pringsheim method) for cultur¬ induced by cultivating the material in ing the Yolvocales and many of the the dark, using a 2 per cent cane-sugar filamentous forms: To prepare, take a solution. The formation of oogonia and gallon battery jar, add 0.5 gram of pre¬ antheridia may be induced by cultivat¬ cipitated calcium carbonate, enough ing in bright sunlight, using a 2 to 4 per humus or garden soil to make a layer cent cane-sugar solution. Sex organs will from y2 to % inches deep, and suf¬ not be formed in partial light or in ficient distilled water to bring the level darkness. within an inch of the top. The culture Cladophora. This alga is found grow¬ jar, covered witli a glass plate, to¬ ing attached to sticks and stones in quiet gether with its contents, is steamed or running water. For cultures, select the (at normal atmospheric pressure) in a forms found in quiet water, and place pressure cooker or Arnold Sterilizer in one or two-gallon aquaria. In larger for an hour on each of two consecutive aquaria Cladophora is likely to grow days before inoculation. (Develop steam too luxuriantly ; even with the smaller slowly to avoid breakage of the battery containers, care must be taken to re¬ jar, and time the operation from the move the excess material from time to first appearance of steam.) The most ef¬ time. fective illumination for this type of culture is provided by two 40 Watt Cladophora is one of the “easy” algae fluorescent tubes located at a six-inch to culture. It can be maintained with distance for the first week and at an so little trouble that pure cultures in increased distance of 18 inches subse¬ nutrient solutions are not usually worth quently for periods of 16 hours daily—in bothering with. addition to normal diffuse daylight. An electric timer (Turtox No. 205A719) Oedogonium. Most species of Oedo- can be placed in the line to switch the gonium are found in the quiet waters of current on and off for the daily illumi¬ ponds and ditches. Forms with dwarf nation period. males are exceedingly small, and appear Spirogyra. Spirogyra is found in the as a fuzzy covering on submerged twigs, field in the quiet waters of ponds, cat-tails, and various other plants. ditches and lagoons, where it often forms Larger filamentous species often form large green mats covering the surface floating mats, which resemble Spirogyra, of the water. Occasionally it occurs in but they are not so slippery. running water, while still less commonly Chamberlain reports that in the case it is found attached to rocks and piles. of Oedogonium diplandrum, “Klebs Spirogyra is not easily cultured for found that a change from a lower to a prolonged periods, but it will sometimes higher temperature would induce the grow well in a balanced aquarium, and production of zoospores. A culture which some species live and increase nicely in nu¬ has been kept in a cold room with a tem¬ trient solutions. Solutions made of dis¬ perature varying from 6 degrees to 0 tilled water containing minute quantities degree Centigrade, when brought into of dissolved commercial fertilizers (such a warmer room with a temperature vary¬ as “Vigoro”) often work well. Spirogyra ing from 12 degrees to 16 degrees Centi¬ cultures should receive plenty of day¬ grade, produced an abundance of zo¬ light and at least some direct sunlight. ospores within two days. Light does not The cultures should be started with dis¬ seem to have any influence on the forma¬ tilled or natural pond water, as city tion of zoospores in this species, but water treated with chlorine or other light is necessary for the formation of chemicals is very destructive to Spiro¬ antheridia and oogonia.” gyra. The related algae, Mougeotia and Although Oedogonium sometimes Zygnema, can be cultured by the same grows well in an ordinary aquarium, if methods that are used for Spirogyra. it is desired to maintain cultures for the entire year, nutrient growing solutions Hydrodictyon is found suspended in are usually desirable. ponds and lakes. When brought into the laboratory it should be placed in an Pleurococcus. Good material for study aquarium which receives an abundance can be secured by collecting pieces ot of direct sunlight; then it should grow tree bark on which this alga is growing. well. It will also grow in some of the Such material may be stored dry ; if nutrient solutions. placed in a moist chamber for 24 hours, Vaucheria. One kind of Vaucheria the Pleurococcus will begin active growth can usually be found in greenhouses, and is then in good condition to study. where it forms a “green felt” on flower It is also possible to grow Pleurococcus pots and damp benches. Such material is on nutrient agar slants. usually in the vegetative condition. An¬ other species, Vaucheria geminata, is Chara and Nitella. Both of these algae often found in ponds and ditches. This are fairly easy to grow in large con¬ species will grow fairly well in labora¬ tainers in the laboratory. Prepare a tory cultures. good-sized battery jar or an aquarium (6-3) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS tank with a couple of inches of sand and Diatoms are often cultured in aquaria pond mud in the bottom and fill with with other algae. Cocconeis, an ellipti- natural pond water. Plant a small cally shaped diatom, is frequently found amount of freshly-collected material in covering Cladophora; Vaucheria is often this and place the container where it covered with smaller forms. will get plenty of light and some direct sunlight. References: Mud taken from the bottom of ponds Chamberlain, C. J. Methods in Plant Histol¬ ogy. University of Chi¬ in which Chara and Nitella are known cago Press. to grow (even when the ponds have com¬ pletely dried up) will often produce ex¬ Johansen, D. A. Plant Microtechnique. cellent new growths. McGraw-Hill 1940. Diatoms are usually found in large Loosanoff& Eagle Use of Complete Fertiliz¬ quantities around springs and in pools ers in Cultivation of Mi¬ and ponds, clinging in great numbers cro-organisms. Science, to filamentous algae, or forming gelatin¬ 95; 487, 1942. ous masses on various submerged plants. Nieuwland, J. A. Hints on Collecting and The surface mud of a pond, ditch or Growing Algae for Class lagoon will always yield some forms. Work. Fresh-water diatoms appear in greatest Midland Naturalist, 1:85, abundance in the spring, are compara¬ 1909. tively scarce in summer, but reappear Pringsheim, E. G. Pure Cultures of Algae again in the autumn. Cambridge 1946. Diatoms show their characteristic movements best when transferred from Smith, Gilbert M. The Fresh-water Algae cooler to warmer water. This phase of of the United States. motility is therefore well illustrated McGraw-Hill 1933. shortly after being brought into thé Ward & Whipple Fresh-water Biology. warm laboratory. John Wiley & Sons, Inc.
Concentrated Media for the Culture of Algae and Ciliates This new and easy-to-use concentrate makes it practical to grow such difficult forms as Volvox, Pandorina and Spirogyra. 61V170 Turtox “Universal” Concentrated Algae Medium. Especially formulated to grow Volvox, Eudorina, Pandorina, Chlamydomonas, Spirogyra, Zyg- nema, etc. Set A; Consists of four units and makes one liter of growing medium in distilled water. Set of four units (distilled water is not included) $3.50 Set B: Consists of four units and makes five liters of growing medium in distilled water. Set of four units (distilled water is not included) 8.50 All prices are f.o.b our laboratories and are subject to change without notice. Algae Cultures Price, per culture, sufficient for 12 students $3.50 Price, per culture, sufficient for 25 students 4.00 Price, per culture, sufficient for 50 students 5.00 Price, per culture, sufficient for 100 students 8.50 Blue-Green Algae *Spirogyra (P.C.) Zygnema *Gloeocapsa *Desmids *Oscillatoria (P.C.) *Scenedestnus (P.C.) *Nostoc (P.C.) * Vaucheria *Plant Plankton (mostly blue-greens) *Diatoms (P.C.) *Eudorina (P.C.) Green Algae (*)—Available at all times mChlorella (P.C.) (P.C.)—Pure cultures *Chlamydomonas (P.C.) 61V55 Living Fresh-Water Algae. An assort¬ *Pandorina (P.C.) ment of three commonly studied species, *Volvox (P.C.) each alga correctly named and in a labeled *Cladophora (P.C.) jar. We reserve the right to select the *Oedogonium species for this assortment, but guarantee to include forms commonly studied and repre¬ Hydrodicfyon (P.C.) sentatives of both the Green and Blue-Green *Pleurococcus (on bark) Algae. Three named cultures, each sufficient Tetraspora for 10 to 25 students $4.75 (6-4) TURTOX SERVICE LEAFLET NO. 59 BASIC LABORATORY EQUIPMENT FOR HIGH SCHOOL BIOLOGY COURSE We offer the following list of equipment suggestions in response to many requests from biology teachers. Those articles marked with an asterisk (*) represent materials which would fulfill the minimum needs of the elementary or high school biology depart¬ ment operating on a limited budget. The complete compilation of equipment is sug¬ gested for groups o,f twenty-four students during a one year course of general biology. It is assumed that standard equipment—such as desks, lights, water and gas outlets—• are available.. Prices are accurate at the time of publication but may be changed with¬ out notice. Charges for transportation (f.o.b. our Chicago laboratories) and special containers are not included. For preserved and living specimens refer to our catalog or write for list of suggestions. IMPORTANT: Write for current price quotations before ordering the materials listed on this leaflet. Microscope Slides Catalog Lot Catalog Lot Quantity No. Material Price Quantity No. Material Price 1 BC1.10 * Type bacteria 81.50 1 E17.17 Cat ovary $1.95 1 BC1.85 * Spirillum volutans .. 1.95 1 E17.12 Rat testis 1.65 1 BC4.2 Typhoid flagella 1 H2.735 * Bone, human, stain 2.50 ground thin 1.95 1 BC4.71 * Bacillus subtilis 1 H3.25 * Muscle, human, spore stain 1.90 striated, smooth, & 1 B1.221 * Volvox 1.65 heart, l.s 2.25 1 B1.252 Desmids .90 1 H2.851 * Human blood 1 B1.256 Spirogyra in con- smear .85 jugation 1.00 1 H2.824 * Frog blood smear .. .80 1 B1.421 * Diatoms .80 1 H10.54 Ear internal 1 B2.322 * Rhizopus. nigri- (Organ of Corti) .. 2.00 cans, zygospores .. .95 1 H10.62 Rabbit eye, l.s. ... 2.00 1 B2.511 Yeast budding .75 1 P5.271 * Taenia, pisiformis, 1 B2.52 * Pénicillium .85 tapeworm 3.00 1 B2.531 * Aspergillus .85 1 P6.43 Necator american- 1 set BWR Wheat rust (3 slides) 3.00 us, male & female, 1 B3.13 * Lichen .85 w.m 3.50 1 B5.816 Fern protnallium .. 1.75 1 P9.6813 Stigmata (spiracles) 1 B5.765 * Fern sporangia .85 of house fly larva .85 1 B6.343 * Male pine cone 1 Z1.21 * Euglena .85 with pollen .80 1 Z1.311 Paramecium .85 1 B7.410 * Stems monocot & 1 Z2.21 Sponge, commer- dicot, x.s 1.35 cial, skeleton .85 1 B7.509 * Leaves, monocot & 1 Z2.31 Grantia, x.s .90 dicot, x.s 1.25 1 Z2.62 Spongilla, gem- 1 B7.3816 Tilia stem, x., rad.. mules, w.m 1.00 & tang.s 1.50 1 Z3.131 * Hydra, l.s .90 1 B7.492 Corn stem, x.s .85 1 Z4.17 Asterias. w.m 1.25 1 B7.210 * Roots, monocot & 1 Z5.ll * Planaria, w.m 1.20 dicot 1.25 1 Z8.41 Earthworm, x.s. ... .95 1 B7.165 Root hairs 1.60 1 Z9.516 House fly, pro- 1 B8.294 Pollen tubes 1.65 boscis 1.00 1 E4.51 * Asterias embryolo- 1 Z9.638 House fly. cornea .95 gy—all stages' 3.00 1 Z9.641 * Insect tracheal 1 E13.78 * Whiteflsh mitosis .. 2.25 system .95 1 E14.39 Frog tadpole, serial x.s. 3.25 Museum Preparations 1 10M282 * Harmful weeds. 1 9D854 Insect wing types herbarium sheets Bio-gram 6.25 (set of 10) 14.00 1 25D151 Slime molds Bio- 1 8M145 Insectivorous plant gram 9.00 set (set of 4) 3.75 1 25D2556 Mushroom Bio- 1 15D102 * Systematic demo- gram 5.50 section collection, 1 25D2721 Puccinia (wheat animal 25.50 rust) Bio-gram .... 6.00 1 2D14 Grantia Bio-gram .. 5.50 1 25D32 Lichen Bio-gram ... 5.50 1 3D1342 Gonionemus Bio- 1 25D3512 Polytrichum moss gram 5.50 Bio-gram 6.00 1 3D422 Pleurobrachia 1 25D3412 Marchantia Bio- Bio-gram 6.00 1 gram 7 25 5D22 Liver fluke 1 25D4102 Fern Bio-gram 5.50 Bio-gram 6.00 1 25D60 1 5D344 Tapeworm (Taenia) Bio-gram 6.00 Bio-gram 6.75 1 8F1 1 8D221 * Earthworm Bio- (complete teaching gram 12.00 unit) 3 00 1 9D1262 * Crayfish Bio- 1 9D677 Honey bee life gram 5.50 history (Riker 1 9D5211 Lubber grasshopper mount) 9.00 Bio-gram 6.25 TURTOX Service Department Copyright, 1958, by TURT0M.Q|UCTS GENERAL BIOLOGICAL SUPPLY HOUSE ( IN CORPORA TED ) 8200 South Hoyne Avenue Chicago 20, Illinois THE SIGN OF THE TURTOX PLEDGES ABSOLUTE SATISFACTION Printed in U.S.A. Skeletons Quan¬ Cat. Quan¬ Cat. tity No. Material Price tity No. Material Price 1 14S10102 Human skeleton & 1 13S61 Turtle $39.50 steel cabinet $319.00 1 13S71 Chicken 46.00 1 13S452 Grass frog , 16.00 1 13S81 Cat 46.00 1 13S352 Perch 22.00 Models 1 TM-1 * Model of typical 1 TM-15 * Model of hydra 35.70 cell 21.00 1 TM-412 Anatomical model 1 set TM-10 Ten models of of human eye .... 55.00 mitosis , 95.00 1 TM-418 Anatomical model i TM-11 * Model of parame- of human ear .... 44.00 cium , 28.75 1 TM-462 Anatomical model i TM-115 * Model of amoeba ... 30.00 of human trunk Models of frog em- with head, sexless 275.00 1 set TM-149 bryology , 97.00 Charts—Qu iz Sheets Catalog No. Catalog No. Key Quiz C.R. Key Quiz C.R. Card Pad Chart Card Pad Chart 1.01 1.01 CR 1 Typical cell 16.113 16.113 CR29.2 Chick development 1.021 1.021 CR 2 Types of animal 17.3 17.3 CR30 Cat dissection cells 18.1 181 CR50.0 Bacteria types 1.031 1.031 CR 3 Animal mitosis 18.2 18.2 CR51 Spirogyra 1.04 1.04 CR 3.1 Spermatogenesis & 18.3 18.3 CR52 Ulothrix Oogenesis 18.4 18.4 CR53 Oedogonium 1.06 1.06 CR 3.3 Animal kingdom 19.1 19.1 CR58.01 Slime molds 1.2 1.2 CR 4 Ameba 19.4 19.4 CR58 Rhizopus, bread 1.4 1.4 CR 5 Paramecium mold 1.42 1.42 CR 5.2 Euglena 21.3 21.3 CR64 Fern life history 2.3 2.3 CR 6 Grantia 24.2 24.2 CR71 Flower, generalized 3.3 3.3 CR 7 Hydra, l.s., x.s. 22.5 22.5 CR67.5 Lily life history 4.1 4.1 CR 9 Starfish 24.3 24.3 CR72 Privet leaf 5.2 5.2 CR10 Planaria 24.25 24.25 CR71.5 Seed dispersal 5.5 5.5 CR 8.4 Tapeworm, 23.1 23,1 CR68 Monocots & dicots Moniezia compared 8.3 8.3 CR11 Earthworm 24.61 24.61 CR73.5 Dicot stem anatomy 9.2 9.2 CR12 Crayfish dissection 24.62 24.62 CR73.6 Monocot stem 9.8 9.8 CR18 Grasshopper anatomy dissection 28.1 28.1 CR90 Six monohybrid 9.85 9.85 CR20.5 Termite life history matings 10.2 10.2 CR21 Clam dissection 28.2 28.2 CR91 Di-hybrid matings 12.3 12.3 CR21.4 Amphioxus dissec- 28.3 28.3 CR92 Independent in- tion heritance 13.02 13.02 CR21.53 Arterial arch circu- 29.3 29.3 CR100 Sex-linked in- lation in vertebrates heritance 13.06 13.06 CR21.55 Lamprey anatomy 13.1 13.1 CR21.6 Shark dissection 13.6 13.6 CR21.50 Vertebrate brains 14.6 14.6 CR23 Frog dissection 48 pads Quiz Sheets (25 per pad) : . $26.00 14.9 14.9 CR26 Frog arterial system 14.12 14.12 CR27.2 Frog, male and fe- 48 Key Cards: . 7.68 male urogenital sys¬ 48 Classroom Charts 17 x 22": tem . 45.60 14.14 14.14 CR27 Frog development 1 390D613 *Easel Chart Stand . 5.00 16.4 16.4 CR29 Pigeon dissection Catalog Lot Quantity No. Material Price 24 385D185 Turtox sexless human manikins $5.00 24 385D13 Turtox earthworm animalkins 5.00
Kodachrome Lantern Slides
Size 2" x 2" Kodachrome 14L365 Bacteria nodules on roots Lantern Slides, each $1.00 35L41 Algae (Biochrome chart) 60L111 Amoeba proteus 52L21 Yeast, sporulating 60L312 Paramecium caudatum, fission 7L24 Arcyria, slime mold 60L313 Paramecium caudatum, con¬ 53L122 Physcia, detail of cup portion jugating 7L77 Cladonia pixidata, with cups 16L017 Grantia growing on seaweed 7L381 Coprinus atramentarius, 16L026 Physalia pelagica cluster 16L033 Astrangia danae 8L11 Marchantia, female heads 61L211 Hydra, entire with bud with young sporophytes 61L221 Obelia, entire colony 8L416 Mnium, large patch with 16L0732 Asterias forbesii, feeding sporophytes 16L087 Lytechinus variegatus, sea 8L87 Equisetum arvense, horsetail urchin 8L55 Polypodium hesperium 16L25 Nereis virens, sand worm 9L36 Pinus ponderosa, male cones 62L361 Clonorchis sinensis, liver fluke 9L37 Pinus ponderosa, female 62L14 Planaria, x.s. through phar¬ cones ynx 62L621 Trichinella spiralis, in 58L21 Lily anther, x.s. muscle 38L321 Lily, megaspore, mother cell 64L15 Cyclops, with egg sacs 10L651 Lilium flower, dissected 16L413 Callinectes sapidus, blue 57L235 Onion root tip, mitosis crab 57L383 Tilia, three-year stem, x.s. 18L832 Cecropia life cycle 57L431 Corn stem, x.s. 64L83 Insect antennae, types 57L561 Privet leaf, x.s. (59-2) Apparatus Catalog Lot Catalog Lot Quantity No. Material Price Quantity No. Material Price 6 105A10N * Insect nets .nylon .. $25.50 1 320A802 ÿ Section razor .$ 6.95 2 105A20N Water dip nets. 1 320A892 " Razor hone 3.25 nylon 9.90 1 320A96 Interval timer 15.00 1 320A105 Slide making kit ... 18.50 105A46 * Water faucet plank¬ s ton gatherer 2.25 24 325A101 Tripod magnifiers .. 49.80 24 110A15 * Carbon tetrachloride 2 325A4055F Wide-field tube microscopes 114.00 insect killing jars.. 27.50 s 110A115 Cyanide lepidoptera 325A4222 Microscopes with jars 7.50 case 552.00 4 330A36 * Turtox Scope-aid .. 31.80 6 110A30 * Spreading boards .. 9.60 18 1800 110A33 * Insect pins (200 ea. 12 325A72 Universal Clamp-on size) 17.50 Lamps 51.00 1 330A20 Tri-Simplex 250 110A325 * Spreading pins 2.00 150.00 24 * Insect boxes 72.00 Micro-Projector 110A51 1 330A386 Projector for 2x2 4 120A10 V.asculums 60.00 119.50 2 13.90 slides, with case .. 120A20 * Plant presses 1 330A67 Filing cabinet for 200 120A36 * Plant mounting 2x2 slides 4.25 sheets 9.50 1 330A54D Metal tripod screen, 24 130A193 Gitsknives 22.80 glass bead surface. 38.95 1 205A20 * Aquarium, 6 1 330A545B Tripod screen gallon 15.95 carrying case .... 6.90 1 205A465 * Dip tube 1.25 1 335A12 Incubator oven com¬ 1 205A551 * Aquarium net .85 bination 60.95 1 210A1501B * Footed terrarium .. 8.25 1 335A342 : Steam pressure 1 210A17 Hydroponic outfit .. 9.45 sterilizer 25.00 4 220A111 Entomological 1 335A381A Double boiler 6.00 breeding chambers. 11.60 12 375A895 Test tube racks, 220A4311 Rat cage assem¬ wood 27.00 blies for nutri¬ 335A57 Culture tube tional experiments.. 23.95 baskets 9.30 220A604 Dietetic scale 8.95 24 335A621 Inoculating loops .. 12.10 250A171 * Seed germinating 1 335A7 Blood typing kit boxes 7.00 (80 tests) 9.50 250A40 Fern spore germ¬ 2 335A79 Haemoglobin scales 2.50 inating outfit 2.25 24 350A30 Measuring slides 5.00 24 308A511 * Dissecting sets 68.40 1 325A91 ! Frog holder (for 1 lb. 310A56 * Dissecting pins .... 3.70 circulation demon¬ 2 308A181 * Sharpening stones .. 3.20 strations 1.75 4 308A24 Large forceps 12.00 1 350A75 Respiration appa- 1 308A36 Bone shears 10.00 13.95 1 308A38 Bone saw 10.50 12 350A511 1 Photosynthesis light 1 308A541 screens 13.50 Instructor’s dis¬ ! secting set 10.75 1 375A11 Harvard trip 1 308A60 Steel cabinet for balance 25.00 small instruments .. 17.00 1 375A411 Bunsen burner 1.35 24 310A5310 * Dissecting pans, 1 375A81 Tripod support 4.00 waxed 62.50 12 375A31 Test tube brushes .. 2.75 1 310A51 Dissecting pan, 20 ft. 375A64B : Rubber tubing, large 4.75 3/16” 2.20 24 310A535 * Aluminum dissect¬ 20 ft. 375A64C : Rubber tubing, ing & utility pans 16.50 1/4” 2.80 1 gal. 310A547 Formalin Fume- 2 lb. 375A655 : Rubber stoppers lock 1.95 assorted 3.70 4 oz. 310A5475 Deodorant 2.25 100 375A711 Corks, assorted .... 2.95 100 310A57 Waterproof tags . 1.75 1 375A80 * Ring stand 7.25 24 310A625 Plastic specimen 1 375A885 Aspirator 9.75 bags . 9.90 2 375A892 Beaker tongs 3.50 4 oz. 310A545 Kerodex Barrier 24 380A20 Drawing paper pads Cream (K71 for (20 sheets each) .. 12.00 formalin) 1.65 24 380A30 Drawing pencils, 2H 4.00 1 308A81A Utility cart . 32.95 24 380A30 Drawing pencils, 4H 4.00 1 320A82 Hand microtome . . 49.50 1 pkg. 420A11C * Filter paper, 25cm. 1.50 Glassware and Plastic 12 315A109E * Pyrex beakers, 4 dz. 320A50 * Dropping pipettes .. 2.00 250cc 6.60 1 315A9310 Bell jar 9.75 1 350A72 * Osmosis apparatus .. 6.50 1 gr. 320A10 * Microscope slides .. 3.15 4 315A330 Graduates, assorted 24 320A17 Hanging drop slides 3.40 10cc., 50cc., 250cc., 320A201F * Coverglasses, 22mm. & lOOOcc 22.60 sq. No. 2 4.50 2 315A229C&E Pyrex Erlenmeyer 1 315A25C Filtering flask with flasks, 250cc., & side tube, 250cc 1.85 lOOOcc 2.05 1 376A24 Clinical thermom- 2 315A921 Desiccators 6.20 eter 1.75 1 315A17D Bunsen’s funnel .... 2.75 4 376A23 * Laboratory ther- 24 315A40D * Petri dishes 17.50 mometers 9.60 12 320A64A Culture bowls. 48 315A55A * Specimen jars, (fingerbowls) 13.75 3/4 oz 4.00 24 320A63 Stacking watch 24 315A55B * Specimen jars, glasses 8.50 3-1/8 oz 3.20 1 315A558 * Storage jar, 3 gal. .. 3.95 48 315A55C * Specimen jars, 8 oz. 8.00 1 315A5275A * Storage pail. 48 315A55D * Specimen jars. Polyethylene 3.00 16 oz 10.00 1 gr. 315A82C * Test tubes 6x%’’ .. 11.92 24 315A5D * Display jars, 8 oz. 6.00 4 lb. 315A96C * Glass tubing, 24 315A5E * Display jars, 16 oz. 7.00 7-8mm. O.D 7.00 24 315A5F * Display jars, 30 oz. 11.00 4 lb. 315A96D * Glass tubing. 24 315A585F * Clear shell vials 10-12mm. O.D 6.00 with polethylene 6 315A55F Culturing jars, stoppers 2.20 1 gal 5.40 12 320A24 Glass marking pen- 24 oz. 315A821 * Plugging cotton, cils, red 2.50 bacteriological .... 3.10 (59-3) Chemicals
Catalog Lot Catalog Lot Quantity No. Material Price Quantity No. Material Price
1 lb. Acetic acid, glacial 4 oz. Wright’s blood C.P $2.90 stain $2.00 1 lb. Acid carbolic, 4 oz. * Methyl cellulose uu U.S.P.. cryst reduce movement
1 88 8 lb. Acid hydrochloric, of protozoa), 10% C.P 2.55 strength 1. 1 gal. * Alcohol iso-Propyl, 2 lbs. Paraffin 53 M.P 1. anhydrous 2.25 5 lbs. * Paradichloro- 5 lb. Calcium carbonate, benzene 3. U.S.P 4.40 23 gr. * Bacto Nutrient Agar 1 lb. Ether. U.S.P 1.80 (Bl) to make 1 lb. * Formaldehyde. 40% .40 1 liter 2.00 1 lb. * Glycerine. U.S.P. .. 1.35 2 vi. 420A21 * Litmus paper, red .. .30 4 oz. Iodine, U.S.P., 2 vi. 420A21 * Litmus paper, cryst 2.15 blue .30 4 oz. Potassium iodide, 2 vi. 420A21 * Litmus paper. U.S.P., cryst 1.50 neutral .30 1 pint Xylol, pure .55 1 vi. 420A301 P.T.C. Tasle-tesl 4 oz. Benedict's solution papers 1.00 qualitative .90 1 vi. 420A31 * Potassium iodide— 1 lb. Bouin's fluid 2.75 starch paper .25 Eosin solution. 1 380A78A Turtox Plastic aqueous 1% 1.00 Embedding kit 6.50
N. D. E. A. Most of the biological materials, equipment and teaching aids authorized for purchase under Title III of the National Defense Education Act are described in the Turtox Catalogs. The special 64-page catalog of Turtox Dependable Biological Supplies will be es¬ pecially helpful. This catalog lists the teaching aids most generally used in beginning courses in Biology, Botany and Zoology.
BIBLIOGRAPHY Bulletin 1952, No. 9. “The Teaching of General Biology in the Public High Schools of the United States.” Bulletin Mise. No. 17. “Science Facilities for Secondary Schools.” (For information concerning the above and other free or low cost publications, address Office of Education, the United States Department of Health, Education and Welfare, Washington 25, D.C.) AMERICAN ASSOCIATION OF SCHOOL ADMINISTRATORS. American School Buildings, Twenty- Seventh Yearbook, Washington, D.C., The American Association of School Administrators, 1949. BEUSCHLEIN, MURIEL, and SANDERS, JAMES M. Storage of Classroom Visual Materials, School Science and Mathematics, 50:402-404, May 1950. BURSCH, CHARLES. Providing Appropriate Housing for Schools. Chapter VII of the 45fh Year¬ book, Part I of the National Society for the Study of Education. Chicago, The University of Chicago Press, 1946. BUTTERFIELD, FRANCES WESTGATE. Teaching Nature in New York's Science Centers. Amer¬ ican School Board Journal. 113:23, October 1946. BYERLEY, J. ROY. Planning and Equipping the Science Laboratory. American School Board Journal, 98: 59-62, January 1939. CAMPSEN, HERMAN M., Jr. Purchase, Construction, and Use of Science Laboratory Apparatus. The American School and University, Seventieth Edition, 1945, p. 382-386. COLEMAN, H. S. ed. Laboratory Design. New York, Reinhold Publishing Corp, 1951. 393 p. Educational Exhibits, How To Prepare and Use Them. Washington, U.S. Government Printing Office, 1948. U.S. Department of Agriculture, Miscellaneous Publication No. 634, January 1948. ENGELHARDT, N. L. Changes in Secondary School Buildings. School Executive, 68:25, Aug. 1949. ENGLEHARDT, N. L., Sr., ENGLEHARDT, N. L., Jr., and LEGGETT, STANTON. Planning Secon¬ dary School Buildings. New York, Reinhold Publishing Corp., 330 42nd Street, 1949. GREENE, CLARENCE W. Master Lists and Storage of Equipment Used in High School Courses in Science. American School Board Journal, Part III, Biology, 113:47-48, November, and 48-49, December 1946. HEISS, ELWOOD d., OBOURN, ELLSWORTH S., and HOFFMAN, CHARLES W. Modern Science Teaching, Chapter 11, The Science Classroom and Laboratory. New York, The Macmillan Company, 1950. JOHNSON, PHILLIP G. Science Facilities for Secondary Schools. Washington, U.S. Government Printing Office, 1956. (U.S. Department of Health, Education, and Welfare; Office of Educa¬ tion, Bulletin Mise. No. 17.) McLEOD, JOHN W. Storage Cabinet Assemblies as Dividing Partitions. American School and University, 21st Annual Edition, p. 223, 1949-50. MARTIN, W. EDGAR. The Teaching of General Biology in the Public High Schools of the United States,U. S. Government Printing Office, 1952. (Federal Security Agency, Office of Education, Bulletin 1952, No. 9.) MEISTER, MORRIS. Science Rooms for Secondary Schools. The Science Teacher, 15:75-76, 87, April 1948. (59-4) TURTOX SERVICE LEAFLET No. 2
PRESERVING ZOOLOGICAL SPECIMENS NARCOTIZATION, FIXATION AND PRESERVATION A list of laboratory specimens is published in this leaflet with individual references to one or more narcotizing agents, killing or fixing agents and preservatives. Two tables are supplied: Table No. 1 outlines the recommended narcotization procedures and Table No. 2 furnishes the formulas for general fixatives and preservatives—including those specifically referred to in the list. Where no specific mention is made of a killing agent, the indicated fixative will serve the purpose. It will be noted in most instances that fixative and preservative are identical. Whenever more than one good fixation and preservation method was encountered for a particular specimen, we have offered a choice of several. Numbers Refer to Numbers Table No. 2 Refer to Table No. 1 Killing & Narcoti- Fix- Preser- zation ation * vation * Protozoa 1, 18, 20 22 From 1 to 2 Epistylis 10 19, 20 10 Opercularia 10 19, 20 10 Spirostomum 11 22 From 1 to 2 Stentor 12 22 From 1 to 2 Vortioella 1, 17 19, 20 10 Sponges, fresh water 2 2 Hydra 2, 14 3 (hot) From 1 to 2 Obelia 16 11 11 Anemones 2, 14 3, 11 11 Campanularia 16 11 11 Gonothyrea 16 11 11 Syncoryne 14 11 11 Tubularia 14 11 11 Actinozoa 14 11 11 Pleurobrachia 14 11 11 Starfish 2, 14, 15 11 11 Sea Cucumber 2, 14, 15 11 11 Planaria 3, 13, 15, 19, 21A 12 10 Flukes 3, 13, 15, 19, 21A 10 10 Microstomum 3, 13, 15, 19, 21A 10 10 3, 11 Ascaris 98°C. H O (1 sec.) 10 10, 2 Nemerteans 3 8 8 Rotifers 1, 8, 9, 11, 19 19, 20, 11 10 Bryozoa 3, 8, 15, 16 10 10 Pectinatella 3 hot (kill) 2 Plumatella 3 hot (kill) 2 Oligochaetes (f. w.) 4, 5 11, 25 10, 25 Lumbricus 7 6, 10 6, 10 Leeches 3, 5, 14 11, 25 10, 25 Daphnia 5, 17, 21A 11 11 Crayfish 2, 11 (kill) 2, 11 Ticks & Mites 2 2 Centipedes & Millipedes 15 15 Insects 2, 15 2, 15 Slugs Boiled, Cool H,0 2, 11 (inject) 2, 11 Snails (acquatic) • 14 11 11 Clams 11 (peg) 11 Dolichoglossus 14 11, 6 11 Salpa 14 11, 6 11 Cynthia 14 11, 6 11 Lamprey 11 (inject) 11 Fish 13 11 11 Grass frogs 13A, 21 2 (drown), 10 inj. 10 Frog Larvae & Eggs 4, 13A 10 10 Salamanders 13A, 21 10 10 Salamander Larvae 4, 13A 10 10 Reptiles 6 (Inject) 11, 2 (drown) 11 Rabbit 5, 6, (21 B-10cc.) Gas or drown 7, 11 Guinea Pig 5, 6, (21B-2cc.) Gas or drown 7, 11 Rat 5, 6, (21B-2cc.) Gas or drown 7, 11 Mouse 5, 6, (21 B-O.lcc.) Gas or drown 7, 11 Large Mammals Gas or drown 11 Vertebrate Embryos 3 2 *Inject body cavities of all larger specimens. (The fixing agent is, in most cases, also the killing agent.)
TURTOX Service Department TURTirMlUCTS Copyright, 1959, by GENERAL BIOLOGICAL SUPPLY HOUSE (INCORPORATED) 8200 South Hoyne Avenue Chicago 20, Illinois THE SIGN OF THE TURTOX PLEDGES ABSOLUTE SATISFACTION
Printed in U.S.A GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
Table No. 1. Narcotization Procedures for Invertebrates and Vertebrates Listed on page one. (To slow down movement for study purposes or to produce relaxation in a contractile specimen as a prelude to killing and fixing. In most cases, the killing agent is also the fixing agent.)
Note: Choices of methods are indicated where more than one number is given. It is often advisable to modify methods in order to produce desired results, and we recommend experimentation to find procedures that fit the particular factors and conditions in your laboratory.
1 Butyn (Butacaine Sulfate). Add 0.1% aqueous sol. (freshly made) drop by drop until desired effect is obtained. 2 Carbon Dioxide. Add ordinary charged water to fluid containing specimens. 3 Chloral Hydrate. Add crystals (or small amounts of 2% sol.) to fluid containing specimens. 4 Chloretone. Use a 1% sol. 5 Chloroform. For aquatic specimens add small quantities to the surface of water in the form of a spray and cover container. Or treat specimens with vapor under a bell jar. For non-aquatic animals use screw-cap jars with impregnated cotton attached to under¬ side of cap. 6 Ether. Use like No. 5, but add small amounts directly to habitat water. 7 Ethyl Alcohol (70%). Transfer worms to a dish of water ,and add the alcohol drop by drop over a period of IV2 hours until the quantity of the added alcohol is 1/10 that of the water. Test by pinching tail. 8 Hanley’s Solution: Water 90 cc., Ethyl Cellosolve 10 ce., Eucaine Hydrochloride 0.3 gm. To every 10 cc. of culture fluid add 1 drop of H S. Repeat at 10 min. intervals until desired effects are obtained. 9 Hydrochloride of Cocaine (1% sol.) 2 parts, Methyl Alcohol (70%) % part, Distilled Water 4 parts. Butyn or Hydroxylamine Hydrochloride may be substituted in strengths V2 as cone. 10 Hydrogen Peroxide (3%). Add to habitat water. 11 Hydroxylamine Hydrochloride. Use 1% sol. 12 Hydroxylamine—Magnesium Sulphate. Make 2 solutions: 1) 5% Magnesium Sulphate, 2) 0.25% Hydroxylamine Hydrochloride. Adjust pH of sol. No. 2 to 6.4. Then add enough dry MgS04 to sol. No. 2 to make 3%. Concentrate organisms in small volume of fluid. Add V2 as much sol. No. 1. When speci¬ mens again exhibit motility, add same amount of No. 1 sol. After 5 min. remove most of fluid and add % this amount of sol. No. 2. 13 M.S. 222 Sandoz (Tricane Methanesulfonate). Use 0.1 gram to 1 gram per gal. of habitat water—acting from 1 to 5 min. A 0.1% sol. in sea water may be sprayed on freshly caught marine spec. 13A M.S. 222 Sandoz. Use 1 gm. per gal. of habitat water. Use 2 to 15 min. for immature forms, several hours to several days for adult forms. 14 Magnesium Sulphate. Add saturated sol. drop by drop. 15 Menthol Cryst. Place a few crystals on surface of water containing specimens. Apply cover to conserve fumes. 16 Menthol and Chloral Hydrate (Galligher). Grind to a paste one teaspoon of menthol crystals with a medium-sized crystal of chloral hydrate and a little water. Place specimens in a minimum of habitat water and add enough of the mixture to form a thin layer. 17 Methyl Alcohol. Add 10% sol. drop by drop. 18 Methyl Cellulose. An inert restrictive medium used only to restrain movements. 19 Strychnine Sulphate 2% sol. 20 Tobacco Smoke. Invert hanging drop preparation over smoke-filled test tubes. Observe under low magnification. 21 Urethane. Pour 5% sol. on specimen. Wash in water when sufficiently narcotized. (Use 0.25% sol. on larval forms.) 21A Urethane 1% sol. 21B Urethane. Use 10% subcutaneously in amounts indicated.
(2-2) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
TABLE No. 2 FIXATIVES AND PRESERVATIVES (Qeneral)
(Numbers Apply to List of Reagents on page 4.) Mise. 2 3 5 6 7 8 9 10 II 12 1 4 Numbers Remarks 1. Alcohol, 50% 5cT #13 50 cc. cc. 2. Alcohol, 70% 30 #13 70 cc. Preservative cc. 3. Bouln's Fluid 20 5 #I7A 75 cc. Gen. Fixative cc. cc. Histology Embryology 4. Carnoy's 5 #13 15 cc. Rapid Action cc. Fixative 5. Carnoy's 15 5 #13 30 cc. Ascaris Ova Fluid No. 2 cc. cc. Insects Avoid Over-Fix. 6. Duboscq - 10 5 #I7B 60 cc. Alcoholic cc. cc. Bouin's 7. Embalming 2.5 86 1.5 10 Fluid cc. cc. cc. cc. Universal 8. FAA 25 T~ Ts #13 25 cc. cc. cc. cc. Fixative Improved by Small % qlyc. Cytology 60 4 #16 16 cc. 9. Fleming's ( Deteriorates) Sol. cc. cc. 95 5 General Fix. 10. Formalin & Preserv. 5% Sol. cc. cc. 90 10 General Fix. 11. Formalin & Preserv. 10% Sol. cc. cc. — General Fix. 12. Gilson's 20 880 4 #13 100 cc. Fluid gms. cc. cc. (60%) #15 15 cc. Preserves 0.2 20 #14 20 gms. 13. Greening ~2 0~ 0.2 40 Green Color gms. Sol. gms. gms. cc. gms. in Plants — 1.5 |#I7 4 gms. For Flagellates 14. Hollande's 2.5 Too 10 ams. cc. cc. cc. #13 175 cc. Improved by !5. Insect 280 60 20 50 cc. small % of Preserv. cc. cc. cc. Glycerin 6 1 #13 15 cc. Insects 16. Kahle's 30" Embryology Sol. cc. cc. cc. #I7A 98 cc. Embryology 17. Kleinenberg’s 200 cc. #20 2 cc. 100 #18 2.5 gms. Nervous Syst. 18. Muller's Fluid cc. #19 1 gm. (Keep in dark) — — 19. Osmic Acid 9 #16 1 cc. Protozoan Fixative — cc. 20. Olney's #15 To replace Nitric Acid (See foot¬ Osmic Acid Fumes note) Fixation 21. Perenyi's 4A — #13 30 cc. Protozoa Sol. 30 (90%) cc. #I5A 40 cc. 22. Schaudinn's 5A 1 #13 10 cc. Protozoa Fluid 40 cc. Add No. II cc. just before use. Use @ 70OC. 23. Tellyesnîcky's 100 T~ #18 3 gms. Bryophyta Fluid cc. cc. Amphîb & Fish Embryos 24. Worcester's 14 200 225 ~25~ Plant Tissues Fluid gms. cc. Protozoa 25. Zenker's 5 100 5 #18 2.5 gms. General Fluid gms. cc. #19 1 gm. Fixative - 26. Zirkle's 2.5~ 2 400 #18 2.5 gms. Mitochondria Fluid gms. gms. cc. (Fix 24 hrs.)
(2-3) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
Preparing frogs in one of the Turtox Laboratories
List of Reagents Used in Table No. 2
1 Ammonium Bichromate, cryst. 12 Glycerine 2 Carbolic Acid, cryst. 13 Isopropyl Alcohol 3 Chloroform 14 Lactic Acid 4 Chromic Acid, 1% aqueous 15 Nitric Acid, cone.* 4A Chromic Acid, 0.5% aqu. 15A Nitric Acid, 10% 5 Corrosive Sublimate, cryst. 16 Osmic Acid, 2% aqu. (DANGEROUS) 5A Corrosive Sublimate, sat. aqu. 17 Picric Acid, cryst. 6 Cupric Acetate, cryst. 17A Picric Acid, sat aqu. 7 Cupric Chloride, cryst. 17B Picric Acid, 1% in 95% ale. 8 Cupric Sulphate, cryst. 18 Potassium Bichromate 9 Distilled Water 19 Sodium Sulphate, cryst. 10 Formalin, full str. 40% 20 Sulphuric Acid, cone. 11 Glacial Acetic
*01ney’s Method: To y2 in. of Nitric Acid in small ground stoppered bottle, add a (Turtox News, piece of copper scrap. Invert hanging drop over fumes for 20 Jan. 1953, p. 29) sec. Rinse spec.
References:
Bolles Lee’s The Microtomist’s Vade Mecum. Blakiston, 1950. Peter Gray The Microtomist’s Formulary and Guide. Blakiston, 1954. (2-4) TURTOX SERVICE LEAFLET No. 19
SPECIAL PROJECTS FOR BIOLOGY STUDENTS So many projects are available for great factor in forwarding interest in high-school students who are interested other projects which follow. in Biology that this leaflet can do little (2) One of the most interesting proj¬ more than list a few of the numerous ects for a biology class, and one in which possibilities. Most of the suggestions are any number of students may participate, of a general enough nature to prove suit¬ is the making of a biological survey of able for schools in all sections of the the region in which the school is located. country, and many can be carried on by Field work of this kind, when properly students in the smaller high schools directed, has the tremendous advantage where laboratory facilities are rather of making students realize that “biology meager. The resourceful teacher will be is at their doorstep” ; they cease consider¬ able to add greatly to this list and will be ing biology as a dry textbook subject able to stimulate student interest in many when they awaken to the fact that count¬ special projects that are possible because less new and interesting happenings in of certain favorable local conditions. the plant and animal world are taking One point should be strongly empha¬ place within a mile or so of their school. sized. It is highly desirable that every In making a neighborhood biological student taking the biology course should survey one student or a small group of be held responsible for some special work students should specialize on each divi¬ to be done “on his own” with the mini¬ sion such as (a) Birds, (b) reptiles, (c) mum of help from the teacher. The in¬ insects, (d) flowering plants, etc. Rec¬ terested students will ask for such work; ords should be kept of the species iden¬ it should be the first concern of the tified and when possible specimens should teacher to arouse the interest of the less be brought in to the school laboratory. willing students and to get them to select The permanent record should credit each a special project of some kind. A good student as collector or observer of the plan is to prepare a list of fifty or more species reported by him. projects (even though the class enroll¬ (3) The average biology class usually ment is much less than that) and allow has several members who have their each student to make his own selection. minds set on becoming doctors. For these In this way almost every student will students a general survey of local sani¬ find something to suit his tastes and in¬ tation and health conditions makes a terests, and the completion of the various good project. In connection with this the projects during the term or semester will sewage disposal system, water works, etc., sustain class interest and create a great may come under observation, as well as amount of wholesome competition. studies of various other measures taken for the protection of health. General Projects In making such a survey, valuable help can often be obtained from local officials. (1) TThe record books of the city hall can alsohere might be some question as to the classification of a Biology Club as contribute to the work by showing what a project for students, yet it seems to us ordinances have been passed for the pro¬ that this classification is quite logical be¬ tection of health. These will include, not cause, after all, it takes work to carry on only such items as regulation of sewage a successful club and by far, the greatest disposal, but also measures to guard amount should be done by the students against accidents. themselves. By all means, if your school does not have an active club now, plan Field Projects to organize one within the near future. Turtox will gladly send you a copy of our (1) In connection with the local bio¬ Service Leaflet No. 57 on “The Organiza¬ logical survey mentioned above it should tion and Activities of a Biology Club.” be possible to make good systematic col¬ We give this project first place because lections of such forms as insects, fishes, it is of prime importance and will be a amphibians, reptiles, ferns, fungi, flow-
TURTOX Service Department Copyight, 1960, by GENERAL BIOLOGICAL SUPPLY HOUSE (INCORPORATED) 8200 South Hoyne Avenue Chicago 20, Illinois
THE SIGN OF THE TURTOX PLEDGES ABSOLUTE SATISFACTION Printed in U.S.A. GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS ers, etc., for the school laboratory. Each Living Material Projects specimen should bear a label showing col¬ lection number, locality, date, collector (1) Rearing insects in the school labo¬ and remarks. Refer to Turtox Service ratory is possible in many instances and Leaflets 2 and 3. such experiments enable the class to see all stages of the life cycles living under (2) Identifying of trees in winter and approximately natural conditions. Good the making of a collection of twigs in subjects for such projects are the meal¬ their winter condition. worm (a beetle), drosophila or fruit fly (a true fly), silkworm moth (a typical (3) Identifying of trees in summer moth), and the cockroach (a roach). and the making of a collection of pressed Refer to any of the better texts on and mounted leaves. Entomology and to the following Turtox Service leaflets: No. IS, “Rearing the (4) Bird study offers many possi¬ Silkworm Moth.” No. 1-5, “The Culture of bilities for the entire class as well as for Drosophila Flies and Their Use in Dem¬ individual students. In the spring a “mi¬ onstrating Mendel’s Law of Heredity.” gration calendar” should be kept, such a record showing (a) name of bird, (b) date first seen, (c) locality, (d) date of (2) Maintaining and studying the in¬ greatest abundance, (e) date last seen habitants of a balanced fresh-water (if the bird merely passes through as a aquarium. In this connection remember spring migrant), (f) remarks. Other that a perfectly balanced aquarium worthwhile phases of bird study include (without fish or other large animal in¬ the studying of nests in winter, making habitants) can be maintained in a con¬ and putting up of bird houses, maintain¬ tainer as small as a quart mason jar. ing of feeding stations during the winter In a quart jar Cyclops, small snails and months. countless other minute examples of pond life will thrive for long periods of time. Write for leaflets, etc., to the National Refer to Turtox Service Leaflet No. 5, Audubon Society, 1000 Fifth Avenue, “Starting and Maintaining a Balanced New York City. Fresh-water Aquarium,” and to “Guide to the Study of Fresh-water Biology,” (5) Herbarium specimens are valu¬ by Needham. able teaching aids, and students should be encouraged to bring in specimens for (3) Rearing of protozoa. Students will the permanent school collection. In addi¬ be interested in starting cultures of Par— tion to pressed and mounted specimens amecia, Euglena, etc., and in studying of flowering plants, ferns, etc., pressed the protozoa found in stagnant pond tree leaves and dried specimens of the water. Refer to Turtox Service Leaflet woody fungi should be included in any No. 4> “The Care of Protozoan Cultures well arranged herbarium. Refer to Tur¬ in the Laboratory,” and to “Living Speci¬ tox Service Leaflet No. 24, “Preparing mens in the School Laboratory” (Price and Caring for a Herbarium Collection.” $1.00). (6) Students who live on farms or (4) Rearing of Daphnia to use for who have backyard gardens should be feeding the fish in the laboratory aquaria. encouraged to try a little simple work in Directions are given in Turtox Service plant breeding. Crossing different varie¬ Leaflet No. 23, “Feeding Aquarium and ties of corn, beans or peas is not difficult Terrarium Animals.” and often produces interesting results. (5) Growing and studying molds. Bulletins on this subject may be secured from the United States Department of Place some moist bread under an in¬ verted finger bowl. When does the mold Agriculture at Washington, D. C. first appear? What is it? How did it (7) Collecting cocoons of various originate? Drawings and notes should moths and the egg cases of spiders for be made each day. Microscopic examina¬ study in the laboratory. Best time is tion will be necessary. Refer to Turtox early autumn. Service Leaflet No. 32, “The Culture and Microscopy of Molds.” (8) In the early spring there are few things of more interest than collecting (6) Bacteria. Fill four petri dishes frog eggs. Bring them to the laboratory (or saucers) with culture media of and place them in an aquarium where the nutrient agar (ordinary gelatine will rapid development into young tadpoles do fairly well) and proceed as follown: can be seen by all members of the class. Cover one dish with a clean sheet of Frogs (in the Chicago region) usually stiff paper and place it aside immedi¬ lay their eggs about April first. Toads ately. Touch the tips of the fingers to lay about June first. the surface of the gelatine in the second
(19-2) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS dish. Cough into the third dish. Scatter a Study” and “The Preparation of Birds little street dust onto the gelatine in the for Study.” fourth dish. Label each dish, according (7) Students who are interested in to the above procedure, and when bac¬ biology and who have even average abil¬ terial growth appears note where it is ity in drawing can make charts which most abundant. Why? What conclusions will be useful teaching aids for the should be drawn? A microscopic exami¬ teacher. Refer to Turtox Service Leaflet nation will not be necessary. No. 26, “Making Biology Charts.” (7) Establishing and caring for terra¬ ria typical of different ecological groups. (8) The making of lantern slides of Refer to Turtox Service Leaflet No. 10, biological subjects is most interesting and “The School Terrarium." worthwhile. In many schools students are used for preparing a great number (8) Seed germination offers many in¬ of lantern slides. Under the guidance of teresting experiments. Among the prob¬ the teacher, students will experience no lems wortli considering are (a) time for great difficulty in following directions germination of different seeds, (b) tem¬ given in Service Leaflet No. 45, “Lantern perature effect on germination, (c) mois¬ Slides Any Teacher Can Make.” ture effect on germination, (d) percent¬ age of fertility, (e) contents of “grass (9) Ants are remarkably interesting seed” mixtures, etc. creatures and are easily collected almost everywhere. Study and keep a record Projects for the Laboratory of the activities of a colony of ants in an observation nest. Refer to Turtox Service (1) Regeneration experiments with Leaflet No. 35, “Studying Ants in Ob¬ living planaria (flatworms). Refer to servation Nests.” Turtox Service Leaflet No. 16, “The Cul¬ ture of Planaria and Its Use in Regen¬ (10) ) Experiments with plant hor¬ eration Experiments.” mones, colchicine and gibberellic acid on seeds, seedlings and growing plants. Re¬ (2) Making skeletons of such animals fer to Turtox Service Leaflets Numbers as frog, turtle, cat, etc. See Turtox Serv¬ 47, 54 and 60. ice Leaflet No. 9, “How to Make Skele¬ tons.” (11) Phptomicrography and other (3) Making permanent microscope phases of photography offer many in¬ teresting projects to the camera-minded slides. See Turtox Service Leaflet No. 8, “Making Microscope Slides of Simple student. Refer to Turtox Service Leaflet Objects.” No. 56 “Simplified Photomicrography.” Write for list of special bulletins and (4) Mushroom spore prints. In the booklets to Eastman Kodak Company, autumn, or at any season when fungi Rochester, N. Y. are fairly abundant, collections of spore prints may be made as follows: Cut off (12) Modeling. Enlarged and scale the cap of the mushroom and lay it, right models of many plant and animal struc¬ side up, on a piece of clean white paper. tures can be made of Permoplast or Cover it with an inverted dish and leave other modeling compounds. Models can for 24 hours. The spores will settle to be carved from blocks of wax or of the paper in the form of a “print” char¬ plaster. acteristic of the shape of the mushroom. (13) Incubate chicken eggs and study The spores are of many colors; mush¬ of developing embryos. Refer to Turtox rooms having white or very light colored Service Leaflet No. 17, “Incubation, Fix¬ spores had best be laid on black paper. ation and Mounting of Chick Embryos.” I'he prints may be made permanent by (14) Make micro-replicas of snow¬ painting them with a thin white shellac. flakes and of leaf surfaces, surface struc¬ (5) Embedding specimens in plastic tures of skin, hairs, etc. Refer to Turtox for permanent museum demonstrations. Service Leaflet No. 31. Refer to Turtox Service Leaflet No. 33, Note. Many additional student projects “Embedding Specimens in Transparent are suggested in the sixty Turtox Service Plastic.” Leaflets. (6) Taxidermy is often of interest to Seasonal listings of projects appear below : boys, and while the killing of birds is (1) September unlawful and should be discouraged, This is usually a busy month for the teacher, but an ideal time nevertheless to interest the students there is no harm in collecting small mam¬ in a series of projects to be started now and carried mals for mounting or making into mu¬ on throughout the school year. The resourceful teacher will suggest many possibilities and permit seum skins. Refer to the two following each student, as far as possible, to select the project booklets which can be purchased from that most appeals to his individual taste. (a) Insect Collections. Let one student specialize the American Museum of Natural His¬ on beetles, another on butterflies, etc., to tory in New York City. “The Capture arouse competitive interest. Collections start¬ ed now can, of course, be continued during and Preservation of Small Mammals for the spring.
(19-3) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
MONTHLY PROJECTS (b) Bird Study, with lists of the autumn mi¬ from an egg, and (4) some other suitable grants. substance. (c) Hebarium Collections. Flowers are plentiful (f) Model in clay certain simple subjects of a and some important families of flowering biological nature. plants not found blooming in the spring are (g) Cast in plaster such things as a clam, a now abundantly represented, notably the small snake, etc. asters, goldenrods, and thistles. (d) Start bulbs and other plants for indoor study (6) February in the laboratory during the winter months. Even in the more northern states, regular (e) Collect clumps of mosses, liverworts, small field trips should be undertaken by the latter part ferns, etc., for planting in the school ter¬ of February or the first part of March. rarium. (a) Run a series of seed germination experi¬ (f) Leaf skeletons. May be made at any time, ments, using some unusual and hard to grow but the fresh leaves should be collected and seeds in addition to the standard textbook pressed now. forms. (g) Wild seeds and fruits. Collect as many kinds (b) Run a series of growth experiments with as possible and arrange them in groups, bulbs and tubers, using the onion, potato, based upon mode of dispersal. sweet potato, and dahlia. (2) October (c) Start a date record of the first appearance Field work should be continued during October of the various spring flowers. Do not overlook if climatic weather conditions permit. There will the trees, some of which—like the silver be ample time for indoor activities when the colder maple and willow—shed pollen very early in weather sets in. the season. (a) Deserted Birds’ Nests may now be brought, (d) Collect and bring into the laboratory, eggs in. Note the widely different types, the vari¬ of Ambystoma and other salamanders. Place ous methods of construction and the great them in a large jar of pond water and record variety of building materials used. the progressive stages of development. (b) Collect snails, aquatic insects and small na¬ (e) Incubate a dozen eggs (hen) in a small tive fishes to place in the school aquarium. electric or gas-heated incubator. Open one (c) Look for the silken egg cases of spiders and egg every other day and preserve the series the large cocoons of the Cecropia, Prométhia, of embryos. and Polyphemus moths. If brought into a (f) Make a series of experiments with the com¬ warm room now, the moths will emerge from mon bacteria. (If standard material is not their cocoons in January and February. available, an ordinary double boiler, porcelain (d) Start an indoor nursery of trees and shrubs, saucers and gelatin may be used.) planting seeds that can be found locally. (e) Mammals. What native wild animals are (7) March still found in your region? Which of these (a) Collect freshly-laid frog eggs from a nearby (and of those formerly found there) hiber¬ pond and place them in a balanced aquarium. nate, and are active throughout the winter (b) Collect samples of pollen from at least a months? , . dozen varieties of flowers and examine each (f) Collect and study the seeds of noxious weeds. of these microscopically. Make drawings of Why have these plants been able to spread each kind. so widely and maintain their abundance in (c) Secure small samples of various types of spite of all efforts to exterminate them? local soils (clay, sand, loam, etc). Test for (g) Collect several lots of algae and establish acidity (litmus), for solubility, and examine cultures in battery jars. Examine these with microscopically. a microscope each week. What changes take (d) Put up bird nesting boxes in suitable lo¬ place? calities near the school. (h) Make a series of Protozoan cultures, using (e) Make a special collection of the flowers of hay, rice, wheat, boiled lettuce and meat. the more common shrubs and trees. What forms appear first in each? What (f) Let your students make a natural history follows? survey of the locality near your school. A (3) November considerable amount of friendly rivalry and (a) Grow cultures of at least two common molds. interest can be maintained if one group lists (b) Look up sources of free literature for the the trees, another the herbaceous plants, biology library. Write to Superintendent of another the insects, etc. Documents, Washington, D. C., for lists of (8) April scientific publications. An ideal month for field work. Regular trips (c) Collect and identify leaves (needles) from afield should be taken, with the entire class the coniferous trees found in your locality. attending, (d) Establish a “culture” of mealworms (a (a) Collect and identify specimens of the local Beetle) and study its various life history amphibians (frogs, toads, and salamanders). (e) Make a sky-chart of the November heavens All of these can be kept alive and studied in and learn to locate the large constellations. aquaria or terraria. (f) Secure some natural (not chemically treated) (b) Install an observation hive of honey bees cider vinegar and start a culture of vinegar outside of one of the school windows. eels. (c) Start now (or earlier) the making of an (4) December authentic herbarium collection of the flower¬ (a) Collect and mount for study, examples of ing plants of your region. the winter buds (twigs) of the common (d) Make a series of leaf-prints of trees, using trees of your locality. blueprints or regular photographic paper. (b) Make a series of simple biology charts by the (e) Collect earthworms and study their noc¬ projection-tracing method, using either paper turnal habits. or chart-making cloth. (f) Hatch some silkworm eggs and raise the (c) Make microscope slide mounts of stems, buds, larvae, feeding them young leaves of the etc., by the free hand sectioning method. mulberry or osage orange. (d) Students interested in taxidermy can trap (g) Use Elodea (or some other suitable water small mammals (field mice, wood mice, plant) to demonstrate that green plants pro¬ shrews, moles, etc.) and make up the skins duce and give off oxygen in considerable and skulls for the school study collection. quantities. (e) Take a census of the resident winter birds. (9) May (f) Dip up part of a large ant colony and (a) Collect living toad eggs and study their establish it in an observation ant nest. development in a balanced aquarium. In (5) January what ways do they differ from frog eggs? (a) Look for insect galls on oaks, roses, golden- (b) Make spore-prints of gilled mushrooms. rod stems, poplar, etc. Section these with a (c) Write to the United States Biological Survey, sharp knife and study their structure. Washington, D. C., for information about (b) Bring into the laboratory a piece of solidly the studies made through the banding of frozen surface soil from field or woodlot. our native birds. Place it in a clear-glass battery jar, cover (d) Make photographs of flowers, ferns, etc. for the top with a glass plate and place upon use as lantern slides. Local pictures are a warm, sunny windowsill. What changes always of more interest than those secured take place? How soon? What plants develop in other ways. and grow? What insects or other animal (e) Make a survey of the common household life appear? pests of your community. Secure information (c) Supplement the school collection with needed about the eradication of them and pass this lantern slides made with etched glass or along to any interested families. cellophane. (f) Label the trees of your general locality so (d) Grow some fern prothallia, germinating fern that the general public will take an interest spores on porous earthenware. in them. On the label, give some informa¬ (e) Perform a series of simple osmosis experi¬ tion as to the common name, scientific ments, using (1) parchment paper, (2) name, commercial value, whether native or animal bladder, (3) the inner membrane introduced, etc. (19-4) TURTOX SERVICE LEAFLET No. 12
DEMONSTRATION AND DISPLAY MATERIALS
Most teachers have neither the time museum” have no place in modern edu¬ nor tile special skill necessary for the cation.) creation of large display collections. But Most dry specimens require little pre¬ nearly every teacher occasionally wishes liminary preparation except thorough to save some particularly interesting drying. Needless to say, partially dried specimen and the purpose of this leaflet specimens which still contain considera¬ is to suggest tlie simpler ways of prepar¬ ble moisture should never be mounted or ing and mounting specimens for display stored in air-tight containers. First, dry or demonstration purposes. them thoroughly and if speed is desir¬ There are, of course, countless ways of able, use a low-temperature oven or sev¬ preparing display collections and of eral infra-red heat lamps. Some marine mounting plant and animal specimens specimens, such as starfish, are likely to for demonstration in the school labora¬ retain an unpleasant odor even after tory. Some of these methods (taxidermy, drying. This can be prevented if such for example) require careful study and specimens are first soaked for twenty- long practice; such techniques will not be four hours in 70 percent alcohol or in 10 discussed here, for the interested teacher percent formalin, then rinsed in water can refer to many available books deal¬ and dried. Soaking starfish and sea ing with museum methods. Other meth¬ urchins in saturated borax solution be¬ ods deal with definite types of specimens fore drying will protect sucK specimens or specialized techniques (making insect against destruction by museum beetles collections, mounting herbarium speci¬ and other insects. mens, preparing skeletons, and embed¬ Small crayfish, crabs and similar ding specimens in transparent plastic) ; crustaceans may be preserved by soak¬ and these methods will not be discussed ing them in a saturated solution of borax here, for they have already been covered and water. Make openings in all the in oilier Turtox Service leaflets joint membranes and then place the The ordinary specimens which the aver¬ specimens in the borax solution for twen¬ age teacher may wish to preserve and ty-four hours. Then rinse in fresh water keep for display and demonstration use, and dry in the position in which the fall into two general groups: specimens are to remain permanently. (1) those specimens which can be The wet specimens can be arranged on dried and kept in a dry state, and sheets of cork or balsa wood and pinned (2) those specimens which must be out for drying. After becoming thor¬ preserved in liquid and stored per¬ oughly dry, such specimens are odorless manently in a liquid preservative. and will keep for many years. The origi¬ I. Dry Specimens nal natural colors will not be preserved, but the dry specimens may be varnished Under this heading might be men¬ and tinted with oil paints. tioned such items as seeds, wood and Most dry specimens are best left in bark samples, woody fungi, fossils, shells, their natural condition. However, cut skulls and other bones, moth cocoons, and polished wood samples are improved and such marine animals as the hard by a coat of clear varnish. Some shells sponges, sea-fans, starfishes and sea ur¬ can be preserved and their colors height¬ chins; there are hundreds of others. ened by the application of a very thin ( However, at this point, it must be stated coat of white varnish or colorless lac¬ that no specimen should be saved unless quer. Bones which have been thoroughly it can serve a definite purpose and have degreased and bleached can also be teaching value. The miscellaneous and treated with colorless lacquer, prefer¬ uncataloged lots of plain “junk” which ably sprayed on. Du Pont’s clear lac¬ sometimes occupy valuable space and quer, thinned to water consistency, is continue to be referred to as “the school good for this purpose.
TURTOX Service Department TURTSMOIUCTS Copyright, 1947 by GENERAL BIOLOGICAL SUPPLY HOUSE (INCORPORATED) 8200 South Hoyne Avenue Chicago 20, Illinois
THE SIGN OF THE TURTOX PLEDGES ABSOLUTE SATISFACTION
Printed in U.S.A GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
The storing and mounting of dry dem¬ mounted. Tapeworms, for instance, are onstration specimens is largely a matter most effectively displayed on black glass. of personal taste. Most large specimens To attach the specimen to the glass do not require mounting but if desired plate it is usually most satisfactory to they can be fastened to plaques made of use fairly heavy cotton or linen thread— plywood or heavy cardboard. Smaller white or black. Using a needle, the specimens are best if protected under thread may be passed through the speci¬ glass. Rilcer mounts are suitable, as are men and tied securely at the back of the also the various sizes of partitioned dis¬ plate. Specimens mounted in this manner play cases with glass tops. Such cases usually remain firmly attached to the accommodate specimens having a thick¬ display plate. Printed or written paper ness up to about three inches. Small labels may be attached to the plate by specimens can be mounted with suitable means of Murrayite or any other water- labels on upright glass plates in jars as and alcohol-proof cement. After speci¬ explained below under Liquid Specimens. mens and labels have been attached to the plates and after the plate has been placed in the display jar, the jar should II. Liquid Specimens be filled with liquid preservative. Forma¬ Included in this group are such ani¬ lin solution of eight percent strength mal specimens as sea anemones, spiders, and seventy percent alcohol are the two soft-bodied insects, frogs, salamanders, preservatives most generally used; the snakes, fleshy plants and countless plant formalin solution is cheapest and also and animal forms which are best dis¬ evaporates less readily than alcohol. In played in a liquid preservative. Good either case, the solution should be filtered laboratory dissections, or parts thereof, carefully (to improve clarity) before be¬ are often worth preserving and mounting ing placed in the display jar. in demonstration jars. The jars may now be sealed, either If fresh (living) specimens are to be with a screwed-on metal cap or a ce- prepared for display, they should be mented-on glass cover. carefully preserved. After being killed, This glass plate and display jar meth¬ the specimen should be arranged and od is also frequently used for dry speci¬ pinned out in the desired position in a mens. The procedure outlined above deep wax-lined tray and then covered may be followed except that the speci¬ with preservative. In addition, animals mens can usually be glued or cemented such as frogs, turtles, etc. should have to the glass plate and no preservative the preservative injected into the body is required. Insects and other specimens cavity and into the larger muscles. After mounted in this way are protected from being thoroughly preserved (this usually tlie ravages of museum beetles and other requires several days in the preserva¬ pests if the jar is kept tightly sealed. tive), the specimen may be removed and mounted in a glass jar of suitable size The Care of Demonstration Specimens and shape. Almost any type of clear glass jar It is well to inspect the display or dem¬ may be used. The straight-sided cylindri¬ onstration collection two or three times cal or rectangular museum jars are de¬ each year. A little care and attention sirable, but these jars have been very will often prevent deterioration and pro¬ scarce since the war and are not obtain¬ long the life of valuable teaching ma¬ able as this leaflet is being written. terial. Screw-capped jars of clear glass with Mounted birds, bird skins, mammal straight sides are available in a variety skins, herbarium collections and particu¬ of shapes and sizes, and such jars are larly insect collections are very likely to recommended for school use. They cost be attacked by moths, museum (Der¬ comparatively little and may be sealed mes tid) beetles and other destructive easily and quickly. (The museum jars pests. Such damage can be prevented in usually come with flat glass covers which large measure by keeping such speci¬ must be cemented onto the top of the mens in air-tight containers, or if this is jar.) not possible, in fairly tight display cases. After selecting a jar of the right The latter also serve to protect specimens shape and size, prepare a glass plate to from dust and careless handling. support the specimen you are mounting. The cabinets or other containers in This rectangular piece of glass should which such collections are stored should fit as tightly as possible inside the jar, hold small trays or other open boxes so that it will remain in an upright posi¬ filled with a good fumigant or insecti¬ tion. It may be cut from (a) transparent cide. One of the best is paradichloroben- window glass, (b) opaque white glass or zene, known by the trade name of “Di- (c) opaque black glass; the choice de¬ chloricide.” This actually kills insects and pends upon the type of specimen being their eggs. Moth balls and naphthalene
(12-2) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
Dry corals displayed in a deep, glass-covered display case. Cases of this type, with or without partitions, are available in many sizes and in depths up to about three inches. They are suitable for displaying many kinds of dry specimens.
Mounting specimens on white plates in museum jars.
(12-3) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
Various ways of mounting demonstration specimens. Display cases, riker mounts, glass boxes, screw-cap jars and sealed museum jars are shown in this group. flakes may discourage insects, but will each year for possible leaks and evapora¬ not kill them. The Dichloricide evapo¬ tion. Even carefully sealed jars some¬ rates when exposed to the air; therefore, times develop leaks. Jars containing al¬ the supply must be renewed every few cohol should be watched with special months. care. Bones and other skeletal preparations If the collection becomes badly infest¬ often become dirty and discolored when ed, heroic measures are usually indicated. exposed to dust and handled frequently. The infested specimens can be placed in Such preparations can be cleaned by tightly closed containers with a quantity washing (preferably merely rinsing) very of (a) Dichloricide or (b) carbon bisul¬ carefully in soapy water. If after such phide and left there for several days. washing, they remain discolored, they This treatment will kill all museum pests. may be bleached in hydrogen peroxide (Caution, the fumes of carbon bisul¬ solution. Bones which are very greasy phide are highly inflammable and explo¬ can be degreased and whitened by treat¬ sive. It must be used with great care). ing them with carbon tetrachloride. Place The Dichloricide is slower, but much the hones in carbon tetrachloride for sev¬ safer to use. eral hours, then rinse in hot water and Museum and display jars containing dry. (Note: Use carbon tetrachloride liquid should be examined once or twice only in a well ventilated room.) 110A513 Parudiclilorobenzeue. The fumes Size No. Size, inches Each Dozen are harmless to humans, but will kill A 8x12x2 * $3.50 $38.50 moths, museum beetles and most other B 12x16x2* 3.75 41.25 insects. The best protection for insect collections. Per pound $0.70 130A381 Compartment Display Case. Five pounds 3.00 Glass top. A durable display case with compartments for separating the vari¬ 110A555 Display Case. Double frame. ous items of a collection placed in it. Glass top. A low priced entomological The partitions are permanently fas¬ case designed primarily for the display tened to the bottom of the case and of pinned insects but equally valuable extend to the top, to prevent the con¬ for other exhibits such as dried plants, tents of one compartment shifting to hereditary preparations, botanical life another, when the case is moved. histories and other material which may be pinned to the bottom of a case. The Size Size, Number of case has a false bottom of tackboard No. inches Compartments Each composition for pinning insects. It is A 8x12x2* 24 $3.00 covered inside with high grade glossy B 8x12x2* 12 2.90 white paper. The outside is covered C 12x16x2* 30 3.80 with durable black paper. D 12x16x2* 12 3.75
Refer also to Turtox Service Leaflet No. 33, “Embedding Specimens in Transparent Plastic.” All prices are f.o.b. our laboratories and are subject to change without notice. (12-4) TURTOX SERVICE LEAFLET No. 43
EMBRYOLOGY IN THE HIGH SCHOOL BIOLOGY COURSE
Written embryology holds little in¬ Schools situated near to the sea coast interest for the beginner in Biology, but may prepare cultures of eggs in the practical embryology along with the laboratory and watch the development theoretical will usually arouse keen in¬ through to the brachiolarial stage. It terest. For this reason it is well to bring will be necessary to dredge in order to into a course in Biology as much work as obtain the young starfish since they will is possible on the study of the develop¬ not metamorphose in the laboratory. ment of living forms, and of prepared In preparing a culture of eggs in order slides. This article is written in order to to study the various developmental give the Biology teacher some ideas stages, eggs should be taken from the which will be helpful in teaching the ovaries of a ripe female and be placed in phase of Biology dealing with develop¬ a culture dish containing an inch and a ment. half of salt water. After forty-five min¬ Field trips should be planned and so utes the water would be changed to fresh conducted that the student will get some salt water, and a drop or two of solu¬ general ideas as to adult life, differences tion from the testis of a ripe male should in sex, environmental conditions affect¬ be added. Fertilization takes place soon ing the laying, fertilization and growth afterwards, and in an hour or an hour of the egg. Comparison of various forms and a half cleavage begins and proceeds in their development is interesting and rapidly. The blastulae will be formed essential to an understanding of embry¬ within twelve hours, and the gastrulae ology. may be seen swimming around the dish within eighteen or twenty hours. Bipin¬ Starfish naria will be formed within three or Starfish development lends itself read¬ four days. Brachiolaria may be found ily to class room study since the earlier after five or six weeks. The water should stages of cleavage are simple and give be changed on the culture each day. the student the basic facts of division. Ascaris Inland schools do not have the advantage For a detailed study of the egg, fertili¬ of using living material, but slides show¬ zation, nuclear processes following ferti¬ ing the various steps in development lization, and the final division of the cell may be used as a substitute. The fol¬ (mitosis), the eggs of Ascaris megalo- lowing stages should be studied and cephala are excellent. As these eggs are attention called to various structures: relatively small and opaque, little can The unfertilized egg; protective mem¬ be observed from the study of whole branes surrounding the egg, cell wall, mounts. Slides showing sections through cytoplasm, nucleus and nucleolus. the uterus, where the eggs are fertilized Early cleavage; the egg divides into and pass through the early stages of two cells of equal size. their development, should be used. Late cleavage; the formation of a clus¬ For a study of cell division, (mitosis in ter of more or less regular cells. animal cells) nothing is better than well Blastula; single layer of small cells prepared sections of the whitefish egg. enclosing a blastocoele. These are even clearer than Ascaris and Gastrula; showing the invagination of are unexcelled for use by high school cells at one pole in the beginning students. (Refer to slide E13.78 listed on formation of the germ layers and page 4.) of the digestive tract. Bipinnaria; early larval stage. Wild Fruit Fly—Drosophila Brachiolaria ; later larval stage show¬ The culturing and observation of the ing the formation of the arms. development of the fruit fly may be car¬ Young starfish immediately after ried on very easily in the laboratory. metamorphosis. Fruit flies can be obtained around places
TURTOX Service Department TURTtpROfUCTS Copyright, 1959, by GENERAL BIOLOGICAL SUPPLY HOUSE (INCORPORATED) 8200 South Hoyne Avenue Chicago 20, Illinois THE SIGN OF THE TURTOX PLEDGES ABSOLUTE SATISFACTION Printed in U.S.A. GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS where over-ripe fruit is found. The sexes the egg from dirt, bacteria and various can be distinguished in that the male has insects which would otherwise prey upon a small, pointed and black-tipped ab¬ them. The egg itself is divided into two domen. The abdomen of the female is regions: the light colored vegetal pole slightly broader, and is finely striped. and the dark animal pole. Several pairs of flies should be placed Cleavage takes place within several in a milk bottle containing part of a ba¬ hours after fertilization, and should be nana. The mouth of the bottle can he studied carefully. Special note should be plugged with cotton. Within a short time made of the planes of division. The first small white eggs will be seen around the cleavage divides the egg into two sym¬ sides of the bottle. With careful observa¬ metrical halves, while the second cleavage tion one may see the females depositing forms four equal cells. Third cleavage eggs. The egg is so small that the cleav¬ is at right angles to the first two and is ages cannot be observed, moreover, early a little above the equator of the egg. cleavage often takes place before the egg After a number of later divisions, the is laid. After several days small white egg comes to consist of a layer of cells larvae will be seen crawling to the bot¬ enclosing the cavity, or blastocoele. This tom of the bottle and feeding upon the stage is known as the blastula stage. banana. In a good culture, hundreds of By the more rapid division of the cells larvae may be seen making their way at the animal pole the blastula becomes through the fruit. As they feed, they converted into a gastrula (or yolk plug) grow rapidly and several days later full- stage. With the formation of the gastrula, grown larvae will crawl up the wralls of the germ layers are differentiated. the bottle. These become more sluggish Soon after the closure of the blasto¬ and turn a crisp brown in color. Soon pore a groove will be noticed extending they stop moving and go into the quies¬ over one surface of the developing egg. cent pupal stage. The young emerge This is the beginning of the neural tube. after twelve days. As they break through Later the groove becomes more pro¬ the cocoon they are of a dusky appear¬ nounced and the sides approach one an¬ ance and not so active, but after several other until they meet, forming an en¬ hours they take on the characteristic color closed tube. The embryo elongates and and actively fly around their container. soon becomes so differentiated that (Living cultures of wild fruit flies, as marked distinctions will designate the well as other strains of Drosophila may anterior and posterior regions. Gill slits be secured at any season from Turtox at develop, the embryo leaves the jelly and $3.50 per culture if you are not able to swims to the surface of the water. The collect the flies locally.) external gills may best be observed in a 6 or 7 mm. tadpole. The appearance of the legs, the growth Frog of the tadpole and the absorption of the The complete development of the frog tail are processes which the student will can be studied in the laboratory. If pos¬ thoroughly enjoy watching. If possible, sible it is well to let the class make a carry some tadpoles through to complete field trip some morning in early spring metamorphosis. when the frogs are laying eggs. The eggs are laid in the early morning, but copu¬ lating pairs of frogs may be found if one Chick is on the lookout for them. Eggs will be A form most interesting to watch in found in clusters lying near the surface its development is the chick. However, of the water. Several “batches” should a bit of technique is required since the be collected in pails and taken to the eggs must be incubated and because they laboratory. Each student should be given are enclosed in a shell. some eggs in order that he may follow In studying the egg, before and during through the development. incubation, crack the shell and allow the For culturing eggs in the laboratory, contents to flow into a finger bowl one- they may be placed in finger bowls half half filled with saline solution. If the filled with spring water (or tap water egg has been incubated the temperature which has stood overnight). Each day of the solution must be 103° Fahrenheit. most of the water should be poured off For incubation, an incubator heated to and fresh water added. If one wishes to 103° may be used, but if the incubator carry the development through to meta¬ is not accessible, a sitting hen may be morphosis, some eggs should be placed in used as a substitute. A nest can be made a balanced aquarium. Three or four days in some quiet corner of the laboratory. after hatching tadpoles begin to feed The following stages should be studied: upon algae, decaying animal tissues, or Unincubated egg—membranes protect¬ almost any type of meat. ing the egg, food content, blastodisc A study of the undivided egg will well established (earlier develop¬ show that it is enclosed in three jelly ment takes place within the oviduct membranes. These membranes protect of the hen.)
(43-2) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
FROG DEVELOPMENT A. Single cell; B. Two-celled stage; C. Four-celled stage; D. Eight-celled stage; E. Sixteen-celled stage; F. Thirty-two-celled stage; G. Many cells; H. Blastula in section; I. Beginning blastopore; J. Yolk plug; K. Gastrula in section; L. Neural folds; M. Early embryo; N. Embryo in long, sec; O. Embryo in cross sec; P. Nine- millimeter tadpole. A large series of Turtox Key Cards, Quiz Sheets and Charts dealing with embryology is available. Write for your free copy of the Turtox Three-Way Check¬ list of charts and biological drawings.
Incubated eggs of: No. 17, “Incubation, Fixation and 13 hours—primitive streak. Mounting of Chick Embryos”). 18 hours—primitive streak and head The development of numerous other process. forms may be studied either as substi¬ 24 hours—several somites have been tutes for the above or to supplement the formed and the nervous system is course. Some suggestions are: sea-urchin, well on its way. crepidula, honey bee, crayfish, crab and 33 hours—heart may be seen, but fish. In order to get some idea as to the circulation has not begun. development of mammals, embryos of 48 hours—circulation well established. rabbit, rat, pig, cat and human are excel¬ 72 to 96 hours—-most organs are well lent. A comparison of the embryos of established. several of these forms is most interesting. If it is impossible to examine the A number of useful texts and refer¬ later development day by day, at least ence books dealing with Embryology are three or four stages between the five- suggested in Turtox Service Leaflet No. day and the twenty-first-day chick 14. should be studied in order that the Some reference charts on this subject student may observe the diminishing of should be available; Turtox publishes the yolk supply, the growth of the many charts and Key Cards on em¬ embryo, and its position within the shell. bryology and these are all listed in the (Refer also to Turtox Service Leaflet free Turtox Three-Way Checklist.
Turtox maintains a stock of practically all materials used in embryology courses. This includes preserved specimens, living material, lantern slides, micro¬ scope slides, models and demonstration preparations. A few of the most commonly used items are listed on the following page.
(43-3) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
PRESERVED MATERIAL FOR Each $0.20 EMBRYOLOGY Dozen 1.00 13C457 Rana pipiens. Four-legged 4C4372 Asterias. Assorted Develop¬ tadpoles with long tails. mental Stages, front single cell Each 25 through gastrula. Recommended for Dozen 1.25 hurried survey by beginning classes 13C458 Rana pipiens. Four-legged and for quiz purposes. Per vial of tadpoles with short tail. mixed stages, enough for ten stu¬ Each .35 dents $3.00 Dozen : . . . . 1.50 4C438 Asterias. Bipinnaria, larval 13C700 Chick blastoderms. Especially stage. Per vial of one dozen. . 3.00 fixed and flattened for mounting. Stages from twelve hours up to one 4C439 Asterias. Brachiolaria, late lar¬ hundred hours after incubation. val stage. Per vial of one dozen 3.00 Please specify stage wanted when ordering. Per dozen of a single 4C4392 Asterias. Minute specimens stage 17.50 immediately following metamorpho¬ 13C79 Chick embryos. Older ages. sis. For whole mounts. We can supply chicks of various Each 35 days’ incubation for the entire Dozen 3.50 21-day period. 13C451 Rana pipiens. Grass frog. Seg- Per single specimen 1.75 mentation stages. 13C799 Chick embryos. Intermediate A. 1-cell stage. Dozen . .90 stages can often be supplied. As¬ sorted stages not accurately B. 2-cell stage. Dozen . .90 graded. 4-cell stage. Dozen . .90 C. Dozen 3.50 D. 8-ccll stage. Dozen . .90 E. 16-cell stage. Dozen . .90 MICROSCOPE SLIDES FOR F. 32-cell stage. Dozen . .90 EMBRYOLOGY G. Early blastula. Dozen... . .90 Set EHS—Beginning Embryology Set H. Blastula. Dozen . .90 Nine slides as listed below. Set $14.20 13C452 Rana pipiens. Gastrula stages. E4.13 Asterias mature eggs, w.m. .85 A. Crescent-shaped blastopore. E4.21 Asterias polar body formation Dozen 90 (maturation complete before en¬ B. Early yolk plug. Dozen. . . .90 trance of spermatozoa), w.m. 1.00 C. Late yolk plug. Dozen 90 E4.31 Asterias, early and late cleavage D. Intermediate stage between late (radial). By use of high power it is yolk plug and early neural plate. possible to see cells in mitosis, Dozen 90 w.m 1.00 13C453 Rana pipiens. Early embryo E4.41 Asterias, blastula (coeloblastic). stages. Stained to show the individual cells. A. Neural plate. Dozen 90 w.m 1.00 B. Neural groove. Dozen 90 E4.42 Asterias, gastrula (embolic). C. Intermediate stage between neu¬ Stained to show the individual cells, ral groove and neural tube. w.m 1.00 Dozen . .90 E4.61 Asterias, bipinnaria (free-swim¬ D. Neural tube. Dozen . .90 ming larval form) w.m 1.10 13C454 Rana pipiens. Early larval E6.24 Ascaris, early cleavage, spindles in all stages of mitosis. An excellent stages. slide to demonstrate animal mito¬ A. Hatching stage. Dozen.. . .90 sis 2.25 B. Gillridge. Dozen . .90 E13.78 Mitosis in egg of whitefish. C. External gills. Dozen..., . .90 D. Early operculum. Dozen. . .90 Every stage of mitosis is shown E. Late operculum. Dozen.. . .90 clearly and in abundance on each slide 2.00 3C455 Rana pipiens. Late larval E14.61 Frog, ten embryological stages, stages. Tadpoles without legs. from one cell to young tadpole. A. 10 to 15 mm. in length. Mounted entire in a depression slide. Dozen 90 All stages on each slide.... 4.50 B. 16 to 25 mm. in length. E16.45 Chick, 33-hour (11-14 somites). Dozen .90 Head fold covers extreme tip of 13C456 Rana pipiens. Two-legged head. Head begins to turn left. First tadpoles. indication of cars, w.m...... 2.25 All prices are f.o.b. our laboratories and are subject to change without notice.
(43-4) TURTOX SERVICE LEAFLET No. 14 A SELECTED LIST OF BOOKS FOR THE BIOLOGY LIBRARY We offer this brief list of books as suggestions only, admitting freely that many excellent texts have been omitted. A wider selection can be found in any good bookstore or in the catalogs of book publishers, Reference should also be made to price lists of pamphlets and books published by the United States Govern¬ ment Printing Office. Note: General Biological Supply House does not sell books, except for the special booklets we publish. Orders for other books should be placed with the publisher or your local bookstore. (Numbers refer to Biology list on page 4. ) Author Title Publisher Price Adell & Welton Lab. Course in Biology with Tests 13. ..$2.76 Bacq & Alexander Fundamentals of Radio-Biology 1. .. 6.50 Baker & Mills Dynamic Biology Today 26. .. 5.20 Boyd Autoradiography in Biology & Med 1. .. 8.80 Bridges & Brehme Mutants of Drosophila Melanogaster 7. .. 1.50 Celeste Biology for Catholic High Schools 2. .. 4.80 Colin Elements of Genetics 24. .. 6.50 Comar Radioisotopes in Biology & Agriculture 24. .. 9.50 Demerec & Kaufman Drosophila Guide 7. .. .25 Dodge-Smallwood-et al Elements of Biology 2. .. 5.25 Fitzpatrick & Bain Living Things 16. .. 4.12 Goldstein Genetics is Easy 19. .. 4.00 Goldstein How to Do an Experiment 14. .. 2.60 Harvey Bioluminescence 1. .. 13.00 Haupt Fundamentals of Biology 24. .. 5.50 Heilbrun Dynamics of Living Protoplasm 1. .. 6.50 Hunter Problems in Biology 3. .. 3.60 Hunter & Hunter Biology in Our Lives 3. .. 4.48 Kamen Isotopic Tracers in Biology i 1. .. 9.50 Marsland Principles of Modern Biology 16. .. 6.75 Needham Guide to Study of Fresh-Water Biology 9. .. 1.00 Pauli The World of Life 17. .. 6.75 Ritchie Biology & Human Affairs 37. .. 4.48 Sinnott-Dunn-Dobzhansky.. Principles of Genetics 24 ... 6.75 Spector (Ed.) Handbook of Biological Data 28 .. 7.50 U.S. Atomic Energy Comm..,Lab. Exp. with Radioisotopes (booklet) 32 .. .30 U.S. Atomic Energy Comm.. Radioisotopes—Uses, Hazards & Controls.... 32 U.S. Atomic Energy Comm.. .Isotopes in Agricultural Studies (pamph.) . . .30 Vance-Barker-Miller Biology Activities 21 1.72 Yance-Miller .Biology for You 21 4.80 Wichterman .The Biology of Paramecium 24 9.50 Williams .The Living World 23 6.25 Woodruff & Baitsell Foundations of Biology 23 6.75 Botany Betsey Morphology & Taxonomy of Fungi 24. 9.50 Cobb A Field Guide to the Ferns 17. 3.95 Conrad How to Know the Mosses & Liverworts. 5. 2.50 Durand .Field Book of Common Ferns 25. 3.50 Emerson Basic Botany 24. 6.00 Emerson & Shields Laboratory & Field Exercises in Botany .24. 2.75 Gray Manual of Botany 3. 12.50 Harlow Trees of the E. States & Canada .24. Haupt Plant Morphology .24. 8.00 Henrici Molds, Yeasts, & Actinomycètes 35. 7.50 Hill-Overholtz-Popp Botany (for Colleges) ,24. 7.50
TURTOX Service Department ftnq- Copyright, 1958, by TURTOX^ROiUCTS GENERAL BIOLOGICAL SUPPLY HOUSE ( IN CORPORATED) 8200 South Hoyne Avenue Chicago 20, Illinois THE SIGN OF THE TURTOX PLEDGES ABSOLUTE SATISFACTION Printed in U.S.A. GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
Hough Handbook of Trees, N.E. States 23...$8.75 J aquès Plant Families—How to Know Them 5... 2.00 Mathews Field Book of American Wild Flowers 25... 5.00 Medsger • Edible Wild Plants 23... 5.95 Muenscher • Aquatic Plants of the United States 9... 5.00 Muenscher • Poisonous Plants of the United States 23... 4.95 Pohl How to Know the Grasses 5... 2.25 Platf • American Trees 10... 3.50 Pool •Basic Course in Botany 13... 5.75 Prescott • How to Know the Fresh-Water Algae 5... 2.25 Smith ■Fresh-Water Algae of the United States... .24... 12.50 Smith •Mushroom Hunter’s Field Guide 34... 4.95 Thom & Raper.... •Manual of the Aspergilli 36... 7.00 Thomas •Field Book of Common Mushrooms 25... 5.00 Wain & Wightman •Chemistry & Mode of Action of Plant Growth Substances 1... 9.50 Wherry .Wild Flower Guide N.E. & Mid. U.S 11... 3.95 Zoology Baker & Wharton An Introduction to Acarology 23... 10.00 Breneman Animal Form & Function 13... 6.25 Buchsbaum Animals without Backbones 33... 8.00 Demerec Biology of Drosophila 35... 13.00 Ditmars Snakes of the World 23... 5.95 Eddy How to Know Fresh-Water Fishes 5... 2.75 Elliott Zoology 4... 7.00 Hegner & Stiles College Zoology 23... 6.90 Holmes The Biology of the Frog 23... 4.90 Hutchinson A Treatise on Limnology 35... 19.50 Hyman Invertebrates; Protozoa through Ctenophora. .24... 11.00 Hyman Lab. Manual for Elementary Zoology 33... 2.75 Jahn How to Know the Protozoa 5... 2.50 Kudo Protozoology 31...10.75 MacGinitie & MacGinitie.. .Natural History of Marine Animals 24... 8.50 Moment General Zoology 17... 7.50 Nichols North American Fresh-Water Fishes 23... 1.75 Pratt Manual of Common Invertebrate Animals... .24... 9.50 Romer .Vertebrate Paleontology 33... 8.50 Stiles Lab. Explorations in General Zoology 23... 3.75 Turtox Ascaris Megalocephala 12... .75 Welch Limnology 24... 9.50 Woodruff .Animal Biology 23... 4.75 Wright Handbook of Frogs & Toads 9... 6.50 Comparative Anatomy Baumgartner ■Lab. Manual of the Foetal Pig 23... 2.50 Bigelow. .Directions for Dissection of the Cat 23. .. 2.55 Calm • The Spiny Dogfish 23... 2.50 Eddy-Oliver-Turner. • Guide to Anatomy Study of Shark, Necturus, & Cat 35. .. 2.90 Horsburgh & Heath .Atlas of Cat Anatomy 29... 1.85 Hyman .Comparative Vertebrate Anatomy 33... 5.00 Kendall .Microscopic Anatomy of Vertebrates 20... 6.00 Romer .The Vertebrate Body 28... 7.00 Sayles .Manual for Comparative Anatomy 23... 4.15 Shumway .The Frog (Lab. Guide) 23... 2.75 Starks & Cutter.... .Dissection of the Rat 29... .75 Entomology Chu .How to Know the Immature Insects 5... 2.50 Comstock .An Introduction to Entomology 9... 7.50 Fenton .Field Crop Insects 23... 6.75 Fernald & Shepard .Applied Entomology 24... 7.50 Holland .The Butterfly Book 11... 12.50 Jaques , .How to Know the Beetles 5... 3.50 Jaques .How to Know the Insects 5... 2.00 Klots .A Field Guide to the Butterflies 17... 3.95 Lutz . .Fieldbook of Insects 25... 3.49 Matheson .Entomology for Introductory Courses 9... 6.00 (14-2) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS Metcalf & Flint.. Destructive & Useful Insects 24..$12.75 Peairs Insect Pests of Farm, Garden, & Orchard... .85. .. 8.50 Ross A Textbook of Entomology 35... 7.75 Swain The Insect Guide 11... 3.95 West & Campbell DDT 8. . . 8.50 Embryology Arey Developmental Anatomy 28... 9.50 Huettner Fundamentals of Comp. Embry, of the Vertebrates 23... 6.00 Lillie • The Development of the Chick 16... 8.50 Lillie & Moore Lab. Outline of Embryol. (Chick & Pig) ... .33... 1.25 Nelsen .Comparative Embryology of the Verteb 24... 9.00 Patten .Early Embryology of the Chick 24... 4.25 Patten .Embryology of the Pig 24... 5.00 Patten .Human Embryology 24... 12.00 Rugh .The Frog (Reproduction & Development).. .24... 5.00 Weiss Principles of Development 16... 7.50 Ornithology Barton..... How to Watch Birds 24... 3.50 Cruickshank Pocket Guide to the Birds 10... 2.95 Peterson .Birds over America 10... 6.00 Peterson.... A Field Guide to the Birds 17... 3.95 Peterson .Field Guide to Western Birds 17... 3.95 Pough .Audubon Bird Guide 11... 3.95 Nature Study Booth How to Know the Mammals 5... 2.50 Burt & Grossenheider A Field Guide to the Mammals 17. .. 3.75 Carr .Handbook of Turtles 9... 7.50 Cockrum .Laboratory Manual of Mammalogy 6... 4.00 Comstock .Handbook of Nature Study 9... 6.75 Gertsch •The Spider Book 9... 6.00 Hausman .Beginners’ Guide to Attracting Birds 25... 2.50 Hillcourt Field Book of Nature Activities 25... 3.95 Kaston , How to Know the Spiders 5... 2.50 Miner Field Book of Seashore Life 25... 7.00 Morgan .Field Book of Ponds & Streams 25... 5.00 Palmer .Field Book of Natural History 24... 8.50 Palmer ,The Mammal Guide 11... 4.50 Schmidt & Davis Field Book of Snakes (U.S. & Canada) 25... 4.50 Smith .Plandbook of Lizards (U.S. & Canada) 9... 6.00 Stebbins .Amphibians & Reptiles of W. North America 24... 8.50 Verrill Shell Collector’s Handbook 25... 4.00 Wright & Wright.... .Handbook of Snakes (2 Vol.) 9... 14.75 Bacteriology—Parasitology Braun Bacterial Genetics . .28. . 6.50 Breed-et al Bergeys’ Manual of Determ. Bacteriology. . .36. .15.00 Buchanan Bacteriology ..23. . 6.50 Burrows Textbook of Microbiology . .28. .11.00 Dubos The Bacterial Cell 15. . 6.00 Irving & Herrick Antibiotics 8. . 6.75 Kelly & Hite Microbiology 4. . 7.50 Levine Introd. to Lab. Tech, in Bacteriology 23. . 4.50 Prescott & Dunn .Industrial Microbiology 24. . 12.50 Rhodes & Van Rooyen Textbook of Virology 36. . 8.00 Sawitz Medical Parasitology 24. . 6.00 Smith & Conant Textbook of Bacteriology (Zinssers’) 4. . 12.00 Stitt-Clough-Branham Pract. Bact., Hematol., & Parasit 24. . 10.00 Turtox Bacteriology Booklet 12. . 1.00 Waksman Soil Microbiology 35. . 7.50 Aquaria & Terraria Innés Exotic Aquarium Fishes 18... 9.75 Innés Goldfish Varieties & Water Gardens 18... 5.50 Mann .Tropical Fish 12... .75 (14-3) GENERAL BIOLOGICAL SUPPLY HOUSE, CHICAGO, ILLINOIS
Mellen Wonder World of Fishes 10.. .$3.00 Mellen & Lanier 1001 Questions Answered About Your Aquarium 10.. . 3,75 Microscope Slide Technique—Histology Conn Biological Stains 36. . . 5.00 Gray Handbook of Basic Microtechnique... 24.. . 6.00 Guyer Animal Micrology 33.. . 5.50 Ham Histology 21. . .11.00 Maximow & Bloom Histology 28.. .11.00 Stiles Handbook of Micro. Char, of Tissues & Organs 24. . . 3.00 Miscellaneous Bear Chemistry of the .Soil 27.. . 8.75 DeVries German-English Science Dictionary 24.. . 6.50 Ellis & Swaney Soilless Growth of Plants 27.. . 6.50 Hall Introduction to Electron Microscopy 24.. . 9.50 Hawk-Oser-Summerson .Practical Physiological Chemistry 24.. . 12.00 Jaeger Source Book of Biological Names & Terms. . .31. . . 5.75 Johnson & Bleifeld Hunting with the Microscope 12.. . .95 MacLeod & Taylor Rose’s Foundation of Nutrition 23.. . 6.00 Pincus The Hormones (Vol. 3) 1.. .22.00 Pray Taxidermy 23.. . 1.95 Russell Soil Conditions & Plant Growth 22.. .10.00 Saunders American Pocket Medical Dictionary 28.. . 3.25 Sharp Fundamentals of Cytology 24.. . 6.00 Shillaber Photomicrography 35. . Thimann-Scharrer-et al. ... Action of Hormones in Plants & Invertebrates 1.. . 7.00 Tiequet Successful Gardening Without Soil 8.. . 3.50 Turtox Laboratory Experiments in Nutrition 12.. . 1.00 Turtox Living Specimens in the School Laboratory. .12.. . 1.00 Turtox Microscopy Booklet 12.. . 1.00 Turtox (Wells) The Collection & Preservation of Animal Forms 12.. . 1.00 U.S. Atomic Energy Comm...Isotopes in Industry & in Physical & Chemical Research (pamphlet) 30.. U.S. Atomic Energy Comm.. Principle of Isotope Utilization (pamph.)... .30.. Williams .Introduction to Chromatography 8.. . 4.00
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(14-4) CHAPTER III
SUMMARY AND CONCLUSIONS
Recapitulation of Experimental»—Experimental evidence indicates that if there is abundant present-day life activity in the classroom, the child is more likely to develop a lively interest in the subject field* Hence, it becomes apparent that the development of appropriate and adequate tech¬ niques in the learning process is essential. Since one of the chief alms of science instruction is to develop in students habits of thought and action inherent in the methods and attitudes of science, there is need in science courses for classroom situations which will be conducive to the development of such habits. Perhaps the best method now known for se¬ curing such an aim is through the medium of the laboratory exercise where problem-solving may and should be the main objective.
Principles and associated regeneration phenomena have been woven into a resource unit for the purpose of providing for the high school biology teacher a lively prospectus. Using the Question-Experiment approach, experiences have been suggested for stressing several of the key ideas in biology, while at the same time providing a stimulus for fascinating the students and challenging their imaginations and thinking. More specifically, questions on various aspects of regeneration have been raised; appropriate animals mentioned as experimental material, and experiments proposed to provide answers to the questions. Terminating the experimental activities is a suggestive evaluation procedure* Such a procedure involves watching $9 60
(by the teacher) the operation skills developed by the students, their
ability to observe and accurately record data, and their desire to propose new experimental situations for verifying other biological principles. Based on the experiences of his own classroom situation, the teacher is to
devise a factual examination to test whether the students have actually grasped what was intended from the experiments.
Summary of Literature,—Numerous studies have been reported on animal regeneration. Since 17U0, when the phenomenon was described in animals, experimental morphologists have explored nearly all facets of the problem.
It is now generally agreed that: 1, Regeneration in animals is not limited to any particular group. Some degree of repair or restoration of damaged or lost parts occurs in
all groups, from the simple protozoa to the complex mammals. There are among both "lower" and "higher” animal groups excellent regenerators and
poor regenerators,
2, In the restoration of cellular elements in animals having lost a portion of their body, the source of the new cells varies among different
groups. In many of the "lower" forms the cells arise from the division of reserve cells which were already present in an undifferentiated state. Among vertebrates ("higher" forms) most of the cells involved in restoring
the lost part result from the de-differentiation of cells originally present in the area near the cut surface, 3, Both the quantity and quality of regeneration can be altered sig¬
nificantly by drastic changes in the environment: changes in temperature, food supply, acidity of alkalinity of the fluid, and other environmental
factors. 61
it. The regenerated poifeion in nearly all instances re-acquired the precise organization characteristic of the original. However, in some cases (for example, in planarians under certain operable conditions) two heads or two tails may form due to the presence of specific gradients of formative materials in the animal. This means that the regenerate will be morphologically unlike the original,
£, The stimulus for regeneration seems to be associated with the wounding process. When the wound fails to heal in a prescribed manner, re¬ generation does not occur. Appropriate treatment of the animal to insure normal wound healing, releases the block to regeneration. Hence, it is now possible for adult animals that have lost their regenerative abilities to be re-initiated in a capacity for some restoration to occur.
Resume of Findings,—The following findings are reported:
1, Regeneration mak^ specific and definite contributions to the fol¬ lowing major principles of Biology which science education agrees are important in science in general education,
a. Growth and repair are fundamental activities for all protoplasm,
b. From the lower to the higher forms of life, there is an in¬
creasing complexity of structure, and this is accompanied
by a progressive increase in division of labor. In all
organisms, the higher the organization the greater degree of
differentiation and division of labor and of the dependency
of one part upon another*
c. Growth and development in organisms is essentially a cellular
phenomenon, a direct result of mitotic cell division. Cells
are organized into tissues, tissues into organs, and organs 62
into systems, the better to carry on the functibns of complex
organisms*
d. All cells arise through the division of previous cells* Cell
division is the essential mechanise of reproduction, of
heridity, and to a large extent, of organic evolution,
e. The environment acts upon living things, and living things
act upon their environment* Since the environment of living
things changes continually, these creatures are continually
engaged in a struggle with their environment,
f. Adult organisms that differ greatly from one another but which
show fundamental similarities in embryological development,
have originated from similar ancestors* Animals resemble each
other more and more closely the farther back we pursue them
in embryological development,
2, Regeneration experiments and demonstrations are possible in the teaching of Biology at the Secondary School level in the following areas:
a* Protozoa
b* Coelenterates
c, FLatworms
d* Annelids
e* Arthropods (Crustaceans)
f* Echinoderms
g* Bony Fishes
h* Amphibians
i. Mammals
3* Curriculum materials are available in the literature for use by 63 the High School Teacher of Biology which will provide adequate background information for the teaching in these areas*
U* Ample diversified materials are available from which it is possible for an alert teacher to construct other Resource Units in this area.
Conclusions .—The following conclusions are offered from this study:
1. Certain organisms are inherently capable of regenerating the
entire body from a tiny fragment.
2. Other organisms are capable of only regenerating parts of the body.
3. There is a distinct relation between the stage of development
and regeneration in higher organisas.
U. Even higher organisms, including man, under certain conditions,
will undergo regeneration.
Recommendations.--In view of the tremendous concern about creating new approaches to teaching courses in biology at the Secondary School level, the writer makes the following recommendations:
1. Teachers of high school Biology should use this Resource Unit*
2. They should prepare one of their own if they should choose
not to use this one.
3. In another study growing out of this one on animal regeneration,
experiments on grafting as a regenerative process should be
conducted. BIBLIOGRAPHY
Books
Barth, L* G, Embryology» Revised Edition, New York! Henry Holt and Company, 1953*
Bonner, J, T, The Ideas of Biology, New York: Harper and Brothers, 1962,
Hickman, C, P, Integrated Principles of Zoology, St, Louis, Missouri! C, V. Moshy Company, 1956,
Milne, L, J,, and Milne, M, J, The Biotic World and Man, 2nd edition, Englewood Cliffs, New Jersey! Prentice-Hall, Inc,, 1958*
Waddington, C, H, Principles of Embryology, London! George Allen and Unwin, 1957.
Weiss, P, Principles of Development, New York! Henry Holt and Company, 1939*
Weisz, P, B. The Science of Biology. New York! McGraw-Hill Book Company, 1959.
Report
Science in General Education, Report of the Committee on the Function of Science in General Education, New York! D, Appleton-Century-Crofts Company, 1938.
Articles
Burnett, R, W. «Vitalizing the Laboratory to Encourage Reflective Thinking," Science Education, XXIII (March, 1939), 299-30U.
Glass, B. "Perspectives! A New High School Biology Program," American Scientist. XLIX (December, 1961), 52U-531.
Mason, J, M., and Warrington, W, G, “An Experiment in Using Current Scientific Articles in Classroom Teaching," Science Education, XXXIX (October, 195U), 299-30U,
McKibben, M. J, "An Analysis of Principles and Activities of Importance n for General Biology Courses in High Schoolsp Science Education, XXXIX (April, 1955), 187-196. 65 Obourn, E. S., Darnell, E. H., Davis, G., and Weaver, E. K. “Fifth Annual Review of Research in Science Teaching,11 Science Education, XLI (Deceniber, 1957), 375-l|ll.
Simmons, M* P. "A Model Lesson in General Science,” Science Education, (March, 1939), 133-136.
Unpublished Materials
Alberty, Harold. "How to Make a Resource Unit." Unpublished Bulletin, College of Education, The Ohio State University, 19UU.
Weaver, Edward K, “How to Make a Resource Unit." Unpublished Compilation, The State Teachers College at Montgomery, Alabama, 19U6 Summer Session.