The effects of arsenamide and pentacresol and anthelminthics on <italic>Ascaris lumbricoides</italic> var. suum

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Content THE EFFECTS OF ARSENAMIDE AND PENTACRESOL AS
ANTHELMINTHECS ON ASCARIS LUMBRICOIDES VAR. SUÜM.
A Thesis
Presented to
the Faculty of the Department of Zoology
University of Southern California
In Partial Fulfillment
of the Requirements for the Degree
Master of Science
by
Edward Laionde Nicholson
June 1951
UMI Number: EP67195
All rights reserved
INFORMATION TO ALL USERS
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a note will indicate the deletion.
UMI
Dissertation F\jblisNng
UMI EP67195
Published by ProQuest LLC (2014). Copyright in the Dissertation held by the Author.
Microform Edition © ProQuest LLC.
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unauthorized copying under Title 17, United States Code
uest
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This thesis, written by
Edward Lalonde HiehoIson
under the guidance of hXfi...Faculty Committee,
and approved by a ll its members, has been
presented to and accepted by the Council on
Graduate Study and Research in partial fu lfill
ment of the requirements fo r the degree of
Master of Science
n.,. May 25. 1951
Faculty Committee
-7^-f
Chat Chairman
TABLE OP COHTEHTS
CHAPTER PAGE
I. INTRODUCTION ....................................... 1
Introduction ........ .. .. 1
Historical Review . . . . . . . . . . . . . . . 2
II. MATERIALS AND METHODS...............................3
Materials . . . . . . . . . . . . . . . . . 3
Methods . . . . . . . . . . . . . . . ........ 7
III. OBSERVATIONS ....................................... 11
Action of Sections . . . . . . . . . . . . . . 1 1
Pentacresol . . . . . . . . . . . . . . . . .13
Arsenamide . . . . . . . . . . . . . . . . . . 1 5
Action of the whole worms and worm sections
in the artificial media . .. ... . . . . .16
IV. DISCUSSION......................................... 18
Artificial Media . . . . . . . . . . . . . . 18
Drug Action.....................................21
V. SUMMARY......................................... 25
VI. LITERATURE CITED ................................ 26
LIST OP PLATES
PLATE page
I. Illustrations of V/onn Sections....................28
Anterior Section Figure A ................... 28
Intermediate Section Figure B . •••••«• 28
Exposed Intermediate Section Figure C • • • . 28
II. Kymograph. Tracings of the Action of Pentacresol
on Anterior Section.
Figures 1, 2, 3 - concentration 1:10...........29
Figures 4, 5, 6 - concentration 1:1000 .... 29
Figures 7, 8, 9 - concentration 1:1000 .... 29
III. Kymograph Tracings of the Action of Pentacresol
on Anterior Section.
Figures 10, 11, 12 - concentration 1:10,000 . . 30
Figures 13, 14, 15 - concentration 1:100,000 . 30
Figures 16, 17, 18 - concentration 1:1,000,000 30
IV. Kymograph Tracings of the Action of Pentacresol
on Intermediate Section.
Figures 19, 20, 21 - concentration 1:10 .. . 31
V. Kymograph Tracings of the Action of Arsenamide
on Anterior Section.
Figures 22, 23, 24 - concentration 1:30 . . . . 32
Figures 25, 26, 27 - concentration 1:300 . . . .32
Figures 28, 29, 30 - concentration 1:3000 . . . 32
V
PLATE PAGE
VI. Kymograph Tracings of the Action of Arsenamide
on Anterior Section.
Figures 31, 32, 33 - concentration 1:100
with sodium glycocholate ............33
Figures 34, 35, 36 - concentration 1:100
without sodium glycocholate .................33
Figures 37, 38, 39 - concentration 1:30
in a weak acid solution ...............33
The depression mark on the timing line
which accompanies all figures denotes
the time in the experiment when the
drug to be tested was added to the
testing chamber.
vl
LIST OP TABLES
TABLE PAGE
I .....................................................20
X I .................................................... 21
INTRODUCTION
It is intended that this paper shall give an
account of some of the new techniques of testing
anthelminthics on the anterior body sections, intermediate
body sections, and exposed intestinal sections of Ascaris
lumbricoides from the pig. Previously, a major part of the
work in testing anthelminthics has been confined to the
whole worm. This paper attempts to elucidate the
paralyzing or exciting effects of Pentacresol, a secondary
amyl"»tricresol, and arsenamide in various concentrations
on each of the above sections and strips.
In the analysis of each section and strip, rather
than the whole v/orm, a clearer perspective of drug action
is apparent. This type of analysis gives a better
quantitative estimate of the efficiency of the drugs
being tested. It shows tha action of drugs on the
different regions of the Ascaris, and in this, one can
see the correlation of drug action to the worm "in vitro".
This may eventually lead to new experimentation and
investigation of better types of anthelminthics. The
techniques undertaken in this investigation are
modifications of the recently published works of Baldwin
(1943) and Baldwin and Moyle (1947).
HISTORICAL REVIEW
The first worker to use the kymograph in the testing
of anthelminthics was Trendelenburg (1915). He thought
that the earthworm was excellent material for use in
testing any new drug for anthelminthic properties. It was
his belief that the earthworm and A. lumbricoides would
react the same to all types of anthelminthics. Stollmann
(1918) did a great deal of testing of new anthelminthics
on earthworms even going into detail on maintenance of the
earthworms in a rich soil medium, which is, of course,
quite different from the environment of Ascaris.
Kymograph testing did not appear again until Rebello,
da Costa, and Rico (1929) presented an excellent series of
papers on the testing of anthelminthics on Ascaris
lumbricoides var suum, Macracanthorhynchus hirudinaceus,
and several species of Taenia. In their work they used
only anterior parts of the worms tested and their working
medium was Ringer’s solution.
Da Costa (1931) expanded on the action of the
anthelminthics of the phenol group and some arsenic
compounds again using the kymograph technique. Lamson et al
(1932) tried to use the kymograph method but gave it up,
preferring to use whole worms. Lamson and Ward (1936)
proved beyond any doubt that the use of earthworms in
3
testing anthelminthics was of no value. Ten of 126
compounds tested, only 6% shov/ed any correlation between
reaction of A. lumbricoides and earthworms. With 58^ of
the compounds all Ascaris were alive after 20 hours while
all the earthworms died within 2 minutes to 6 hours tested
with the same drugs. The remaining 36^ showed no effect
on either the earthworms or Ascaris.
Baldwin (1943) revived the use of the kymograph and
developed the use of different sections of A. lumbricoides
for testing of anthelminthics as to their effects on
different parts of isolated tissue from the organism. He
also condemned the use of earthworms as testing material
for anthelminthics, as: earthworms have a fairly complete
circulatory system; their nervous system is of a more
complex nature; their cuticle is very thin and is open to
quick penetration of many drugs; and the earthworms ’ mode
of life and environment is very different from that of
Ascaris. Baldwin and Moyle (1947) used the kymograph
technique and introduced the use of dorsal muscle strips
along with the other types of sections for testing
an thelminthi c action.
MATERIALS
Large female Ascaris lumbricoides var suum Linnaeus
were used for all types of testing in these experiments.
The Ascaris from the swine are morphologically similar to
the one found in man, but they differ in physiological
traits, as no cross infection can occur.
The storage medium for the A. lumbricoides was
prepared according to the specifications of Baldwin and
Moyle (1947). A stock solution concentrate was composed
of the following chemicals: 75.5 gms. of HaCl; 14.2 gms.
of KCl; 15.1 gms. of CaClg; GHgO; 10.2 gms. of MgClg.GHgO
and diluted to 1 liter. A stock buffer solution viras made
up of the follov/ing compounds: 250 ml. of 0.2 M KH2PO4,
and 21 ml. of N NaOH which was brought up to 1 liter with
distilled water.
One volume, 500 cc., of concentrated stock solution
received one volume, 500 cc. of the stock buffer solution.
This solution was diluted 10 times with distilled water,
increasing the volume to 10,000 cc. This medium was warmed
to 38^0 for use. A gas mixture of 95^ nitrogen and 5%
carbon dioxide (no correction for oxygen) was bubbled
into the medium until a pH of 6.7 was reached.
The working medium or the solution in which the
sections were tested for the kymograph readings was
prepared fresh from dilutions of stock molar
concentrations. The medium was made up of the following
dilutions: 117 ml. of M NaCl; 7.5 ml. of 120 mM. of
NaHgPO^; 17.5 ml. of 120 mM. of NagHPO^. These solutions
v/ere brought up to 1 liter by distilled water. To each
5
liter of the above solution was added the following
solutions: 30 ml. of 120 mM. KCl; 15 ml. of 120 mM. CaClg;
and 30 ml. of 120 mM. MgClg. This medium was warmed to
38°G and 100 rags, of thiamine hydrochloride was added
just before use.
A heart lever with a gravity fall writing tip was
used throughout the experiments. A load of 0.5 gms. was
placed on the worm sections to maintain a fairly constant
tension. The kymograph drum rotated at a speed of 7 mm.
per minute.
The testing chamber consisted of a large 55 ml.
test tube held in a water bath by a clamp and ring stand.
A glass pipette rod was bent to fit the test tube and at
the lower end of the glass rod a solid glass hook was
formed for the attachment of the posterior suture from
the worm section. The glass pipette rod was made secure
by a clamp to the ring stand, and at the upper end of the
glass rod a rubber tube was fastened for the delivering of
the carbon dioxide and nitrogen to the working solution.
Pentacresol, the "drug" tested in this experiment,
contains five compounds probably in diminishing quantities
from 1 to 5. The structural formulas are:
CH3
cKî
CH^
fn
/Vo,
A
OH
V
11
X 3
y 5
6
It is a yellow liquid with an odor of strong cresol. It is
(very) insoluble in water but fairly soluble in a weak
alcohol solution. At the present time this drug is used
commercially as an oral antiseptic.
Solutions of 1:10, 1:100, 1:1,000, 1:10,000,
1:100,000, 1:1,000,000 (in a 3^ alcohol solution) of the
Pentacresol were prepared for testing on the isolated
tissues.
If this secondary amyl-tricresol is applied in
full concentration to an isolated muscle strip of Ascaris
one can see a definite cresol reaction (where the drug
touches the muscle, the muscle turns white as dehydration
occurs).
Arsenamide, the other drug used in this
investigation, is a para-(bis(carboxymethylmercapto)-
arseno)- benzamide. The new trade name for this compound
is "Gaporsolate". It is a light yellow, odorless powder,
not very soluble in water but soluble in a weak alkaline
solution (0.2 N NaOH). It is slightly soluble in a v/eak
acid solution (0.2 N HCl) and fairly soluble in high
percentage alcohols (60-90^). The melting point is
between 158® - 160®C. The chemical formula is:
7
It Is a trivalent compound and should give the maximum
drug action "in vitro", (Clark 1937).
The solutions for testing the drug as to its
an the Imin thi c properties were made up in the following
concentrations: 1:30, 1:300, 1:3,000 in a weak alkaline
solution. A 1:30 emulsion in a acid liquid. A 1:100
alkaline solution v/ith 0.5^ sodium glycocholate added,
and a 1:100 alkaline solution without the bile salt (this
was to check the effects of a bile salt on the
penetration of the cuticle for drug action).
METHODS
Ascaris lumbricoides females were obtained from
a local packing plant and placed in a thermos bottle
containing the storage medium at a temperature around
38°C. On arrival at the laboratory the worms were washed
in tap water to clean them of any intestinal matter
present. After washing, they were placed in a large glass
container holding about 2,000 ml. of the storage medium.
This container was in a constant temperature incubator
wfhich kept the medium at 38°C.
In isolating each section, two sutures were employed.
In this paper they are referred to, as the "anterior
suture" which is tied 0.5 cm. posteriorly from the
anterior limits of the section and as the "posterior
suture", which is tied 0.5 cm. cephalad to the posterior
8
portion of the section. The sutures placed in this manner
give the isolated sections a better contraction and
relaxation period. If the trailing edges (0.5 cm. at each
end) are absent and the sutures are tied tightly, the
movement of the section is retarded.
Four isolated sections from different areas of the
v/orm are used in this experiment. In the next few
paragraphs, each of the isolated tissues will be described
in detail and diagrams of these sections will be found
in plate I.
The anterior section is obtained from the head
region of the worm. At a point 0.5 cm. from the labia, the
"anterior suture" is tied. With the aid of a small metric
ruler, a distance of 2.5 cm. is measured posteriorly from
the "anterior suture" and at this point the "posterior
suture" is tied. At a point 0.5 cm. caudad from the
"posterior suture" the worm is cut and the isolated
section placed in the working medium. The "anterior
suture" is made fast to the kymograph writer and the
"posterior suture" is tied to the hook on the glass
pipette rod. A period of 15-25 minutes is allowed each
section to settle down to a rhythmic pattern before
tracings on the kymograph drum are started. In plate I
(fig. -A) the placing of the sutures for the anterior
section is shown.
9
The intermediate sections are taken from the middle
portion of the worm. The "anterior suture" is placed 3.5
cm. craniad from the genital pore. The "posterior suture"
is tied 1 cm. in front of the genital pore. Each suture
receives an 0.5 cm. trailing edge and the rest of the worm
is cut away. The section is then placed in the working
medium and the same procedure is followed as in the
anterior sections. In plate I (fig. -B) the placing of the
sutures for the intermediate section is shown.
The exposed intestinal sections, plate I (fig.-C),
are cut from the same area as the intermediate sections.
Instead of the sutures closing the intestine, fine glass
capillary tubes are placed at each end of the section. By
this method, the drug will not act on the cuticle alone
but can enter the intestine (a two-fold action). This
section is attached to the kymograph by the same method as
above.
Tracings on the kymograph are carried out for 5
minutes and, if good motion is found in the section being
tested, 10 cc. of the drug being tested is applied. If
drug action is present, a 3 minute tracing is taken. In
this work, the Pentacresol when added, gives instant
action. The Arsenamide also gives this immediate reaction
and the same period of time is employed.
The same drug concentration was tried on three
10
different worms to check individual reaction. This means
that three anterior sections, three intermediate sections,
and three exposed muscle strips were tested for each
concentration of drug used. With this method one can
check the variation of individual strips and worms to the
action of the drug.
OBSERVATIONS
One hundred and twenty worms of more than average
size {25-35 cm. in length) were used for testing. A few
tests were made on worms which had been in the laboratory
as many as four days. Tests on these worms showed that the
recordings or tracings of the worm’s contractions were so
weak that they were of no value to the investigation.
Following this observation no worm was utilized that had
been in the laboratory for more than 48 hours.
A suitable worm was selected from the storage medium,
held horizontally, and dropped from a height of 12 inches
to inactivate it. This treatment leads to a sharp
contraction of the musculature and the worm remains
motionless long enough to apply the sutures and remove
the sections for testing. This operation takes about one
minute after which the section is placed in a large test
tube (55 ml.) containing the working medium which is kept
at 38°C by the thermostaticly controlled incubator. For
each concentration three sections (from the same area but
from three different worms) were cut and placed in the
working medium. A period of fifteen to twenty-five
minutes was allowed for the section to become active and
give steady contractions. If the action was steady, the
section was placed in the testing chamber.
The section in the testing chamber was attached to
12
the writing arm and, if the activity continued, the
writing point was placed on the kymograph drum for tracing.
The movements of the writing point do not correspond
to simple longitudinal contractions and relaxations such
as are observed with a simple strip of isolated muscle.
The principal movements made by the section consist of
arching in the dorsal-ventral plane. On observing the
movement of the section, one sees as each arc is
established that the ends of the fragment or section are
drawn more nearly together so that the overall length of
the section decreases and there is an upward movement of
the tracing point which is recorded on the kymograph drum.
Muscle arrangement of Ascaris is such that contractions
of the section give a series of 8-shaped and C-shaped
patterns.
The anterior section gave the best results in this
experiment. The structure of this section consists of an
outer cuticle, a subcuticular layer, and muscle cells,
each with its protoplasmic element projecting into the
body cavity. The nervous element is composed of: a
central nervous organ (circum-esophageal ring), a dorsal
longitudinal line with a dorsal nerve trunk, a ventral
longitudinal line with a ventral nerve trunk, four lateral
longitudinal trunks with transverse commissures, and nerve
endings in the sensory papillae on the lips. This section
13
also contains a muscular esophagus with the esophageal
glands, and two lateral lines.
The use of the intermediate and exposed sections
gave very irregular reactions, and out of thirty-four
sections, only three gave regular action for the drugs
being tested. This difficulty could arise from the modified
apparatus used in this investigation as compared to
Baldwins* (1943). If the anterior sections gave rhythmic
tracing at the end of a five minute period, 10 ml. of the
drug to be tested v/as added and another period of five to
ten minutes was allowed for the action of the drug. The
actions of the various concentrations of drugs will be
described in detail below.
PENTAÔRBSOL
1. The concentration of 1:10 and its effect on the
section is shown in Plate 2 (figs. 1, 2, and 3).
The action was very rapid with the paralyzing
effect noticed within one minute after the
administration of 10 ml. of the 1:10
concentration. There is a rapid paralyzation of
the section with a fall in amplitude and tone
level. There was no evidence of a blistering
effect on the section from the strong
concentration of this cresol-type drug.
14
2. The intermediate sections which did give good
tracings are shown in Plate III (figs. 19, 20,
and 21). The action of the 1:10 concentration on
the section was rapid and the amplitude and tone
level were reduced within a period of one minute
and paralysis of the section followed.
3. The action of the 1:100 concentration on the
anterior section gave an excitation reaction
followed by a rise in amplitude and tone level.
Plate II (figs. 4, and 5). In one case Plate II
(fig. 6) there was a rise in amplitude but a
drop in tone level. It could be that this
particular action had a longer relaxation time.
4. The anterior section in reacting with a 1:1000
concentration did not fall in amplitude or tone
level. The contractions increased as shown in
Plate II (figs. 7, 8, and 9).
5. The 1:10,000 concentration of the pentacresol had
no effect. Plate IV (figs. 10, 11, and 12).
6. The 1:100,000 dilution of the pentacresol did
give a slight rise in tone level but no other
effects of the drug on the section. Plate IV
(figs. 13, 14, and 15).
7. The action of the 1:1,000,000 concentration
showed no change at anytime on the anterior
15
sections. Plate IV (figs. 16, 17, and 18).
ARSENAMIDE
1. Plate V (figs. 22, 23, and 24) show that the
arsenamide in a concentration of 1:30 (in a weak
alkaline solution) had a definite paralyzing
effect on the anterior strips. The tone level
dropped in all three sections and contractions
ceased, (figs. 23, and 24).
2. The concentration of 1:300 (in a weak alkaline
solution) showed no effect on tone level or
amplitude. Plate V (figs. 25, 26, and 27).
3. Concentrations of 1:3000 (weak alkaline solution)
showed no response on the anterior sections.
Plate V (figs. 28, 29, and 30).
4. A concentration of 1:100 (weal alkaline solution)
did not bring about any type of change in the
movement of the sections. Plate VI (figs. 31, 32,
and 33).
5. Many workers have stated that the use of sodium
glycocholate (a constituent of the bile salts
of most vertebrates) with an antheIminthic
decreases the permeability of the Ascaris cuticle
to drugs. The effect of a 0.5^ solution of sodium
glycocholate (weak alkaline solution) containing
a 1:100 dilution of the arsenamide powder on the
16
sections had no difference in the action on the
section than when a 1:100 concentration without
the sodium glycocholate was employed. Plate VI
(figs. 34, 35, and 36). This is in agreement with
the observations made by Baldwin (1943).
6. The use of a weak acid solution (HCl 0.2 N) which
contained a concentration of 1:30 of the
arsenamide is shown in Plate VI (figs. 37, 38,
and 39).
The arsenamide was an emulsion in the acid solution. All
worm sections did show an increase in their tone level
after the drug was administered in the testing chamber.
In fig. 39 there is indication of a contraction of the
worm section proceeding paralysis. This type of reaction
also follows the action of thymol, a phenol ant helminthic.
ACTION OP THE WHOLE WORMS AND WORM SECTIONS
IN THE ARTIFICIAL MEDIA
1. The incorporation of thiamine hydrochloride into
the medium gives the section a longer relaxation
period. This prolongs the activity of the
sections employed.
2. Nitrogen and carbon dioxide gas (N-95^ and COg-
5^), with no correction for oxygen content, had
no effect on the Ascaris in the artificial medium.
3. Worms in the artificial media gave their best
17
action at a pH of 6.7.
4. The changing of the storage medium every 12
hours did extend the life span of the Ascaris in
the artificial environment.
DISCUSSION
Artificial media
The artificial media used in this experiment for in
vitro life of Ascaris lumbricoides var. suum, is similar to
that first prepared by Baldwin and Moyle (1947). The main
function of these media is to keep the activity of worms
in vitro at a maximum so tests will be made under favorable
physiological conditions.
For short-term physiological testing on Ascaris,
many different types of artificial media have been used.
Hobson (1948) has summarized the considerable literature
on this subject. He states that from the time of Bunge in
1885 to Baldwin and Moyle in 1947 the basic elements have
alv/ays been simple sodium chloride solutions. Also, that in
1901, Weinland used Bunge’s solution (1^ NaCl) and added an
0.1^ solution of sodium bicarbonate and kept Ascaris alive
for 5 days. He then added carbon dioxide to the solution
and the worms in this medium showed life for ten days. Hall
and Foster (1917) kept ^ lumbricoides alive for fifteen
days in Kronecker’s solution (physiological sodium chloride
plus 0.006 gras, per liter of sodium hydroxide) and two
female worms survived for 24 days and one for 26 days in
this solution. He did not change the medium and added only
distilled water to make up for evaporation less. The
temperature range in this work was 27.5^ to 30^C.
19
Rebelle et alia (1929) and Da Costa (1951) used
Ringer’s solution for testing anthelminthie action on
Ascaris» but only for short periods. Hobson (1948) used
Fenwick’s solution which consisted of four inorganic salts:
NaCl-0.80^, KCl-0.02^, CaClg-O.OS^ and MgClg-O.Ol^. Larvae
of Ascaris survived in this medium for five and one-half
days.
Baldwin (1943) made up a saline solution of the
following materials: HaCl 80 gms./liter, KCl 2gms./liter,
CaClg 2 gms./liter, and MgSO^. THgO 1 gm./liter. To 100 cc.
of this mixture was added 10 cc. of 0.05 M phosphate buffer
at a pH of 6.4 and diluted up to 1,010 cc. with distilled
water (or 10 volumes). In this medium the Ascaris were kept
alive for 12 days.
In this investigation the media employed are similar
to the ones of Baldwin and Moyle (1947).
Hobson (1948) states that in an artificial medium
of 30^ sea water. Ascaris have been kept alive for 28 days.
The relative composition of the four media shown
above are given in Table 1.
20
Table 1.
All figures are in m moles
•
Na K Ca
Mg
Cl
^ 4
Penwi ck’s saline 137 3 2 2 148 nil
Baldwin * s saline 136 3 2 0.4 141 0.4
Baldwin and Moyle 130 24 6 5 170 nil
30^ sea water 141 3 3 16 164 8.5
This experiment deals with keeping Ascaris as
whole worms, but the actual testing is done on isolated
sections. Therefore, two types of media were used; a
storage medium for the whole worms and a working medium
for the isolated sections. The working medium as
indicated earlier was made up of inorganic salts
according to the method of Baldwin and Moyle (1947).
This medium is used in the testing chamber with the
isolated muscle sections to allow the surrounding
environment to be as similar to the pig gut contents as
possible and the internal environment of the host. In
Table 2 the composition or similarities of the working
medium. Ascaris body fluid, and the pig gut contents are
shown.
21
Working medium
Table 2.
K Ca Mg
130 25 6 5
124 27 14 6
130 24 6 5
Cl
Ascaris» body fluid 130 25 6 5 53
Pig, gut contents 124 27 14 6 61
Drug Action
In testing anthelminthies the best criteria for
checking their action are the paralyzing and exciting
effects. If an anthelminthie should excite movement in
A. lumbricoides there would be danger of migration that
could cause severe damage to the host.
The use of isolated sections of Ascaris for testing
anthelminthies gives a clear picture of the action of
the drug. The anterior segment with the circumesophageal
nerve ring and six short trunks that innervate the
anterior region is good material for testing
an the Imi nthi c s acting on nervous tissue. The intermediate
and the exposed intestinal sections are used to show
anthelminthie action on the cuticle and the muscle.
Trim (1944) used ligated and non-ligated Ascaris in
testing the penetration of hexylresorcinol. He found that
the non-ligated worms with the use of their muscular
esophagus w uld push the drug out their anterior end. The
ligated worms were penetrated through the cuticle by
22
hexylresorcinol. Detection is based on the well known
azo-dye reaction of phenols, a color reaction taking place.
The intensities are determined by a colorimeter.
Rogers and Lazarus (1948) with the use of radioactive
phosphorus and autoradiographs found that non-ligated
Ascaris took up the radioactive-phosphate more readily
than the ligated worms. Also, in this work,
Nippostrongylus muris in rats took up radioactive-
phosphate in larger amounts when the radioactive
phosphorus was injected intramuscularly than when it was
given orally to the host. Hippostrongylus muris gets its
food by direct attachment to the intestinal wall.
Ascaris takes its nutrient material by moving
freely in the host intestine without oral attachment to
the intestinal wall.
Prom Rogers and Lazarus (1948) observations, it
seems that to get maximum action of an anthelminthie, one
must knov/ the feeding habits of adult nematodes in their
host.
Baldwin (1948) and von Oettingen (1948) gave
excellent accounts of the action of the phenol type of
drug. In their reports it was generally agreed that the
longer the side chain on the phenol ring, the greater was
the anthelminthie activity. Clark (1937) states that most
drugs with the phenol ring structure have an all or none
23
action on the tissue or animal "in vitro". These
compounds coagulate protein at certain concentrations.
Most phenols do not start to coagulate protein until the
concentration is between 1-5^, and is completed when the
concentration reaches 20^. If the concentration is below
1^, coagulation of protein does not occur.
Pentacresol in this investigation did give an all
or none action on the anterior sections and the
intermediate sections when its concentration was 1:10.
Trim (1944) showed that sodium glycocholate and
other bile salts acted on the Ascaris cuticle surface
layer to form a lipoid complex which would inhibit
penetration of an anthelminthie. Baldwin (1943) used
sodium glycocholate in his work and found no decrease in
penetration of the anthelminthlcs. Sodium glycocholate
was used in this investigation and showed no effects on
penetration or action of the arsenamide.
The arsenamide employed was a trivalent compound
which gives the maximum effect l^n vitro* Clark (1937).
This compound in high concentration could have acted on
an enzyme system or a direct adsorption on the surface of
the worm, which did happen in the use of 1:30
concentration. The physiology and biochemistry of the
nematodes have not been advanced far enough to give the
action of this arsenamide on the sections of worms tested.
24
Knov/ledge of drug action is still incomplete*
Better methods for testing the anthelminthlcs must be
found before the real action of these drugs can be known*
SUMMARY
1. A review of the workers who used the kymograph method
in testing anthelminthie action on Ascaris
lumbricoides is given*
2# The composition of a storage medium and a working
medium for testing of A# lumbricoides in vitro have
been described*
3* Descriptions as to length, diameter, and anatomical
structures of anterior, intermediate, exposed
intestinal segments, and exposed dorsal muscle strips
of Ascaris lumbricoides have been stated*
4* The structure and general characteristics of
Pentacresol and Arsenamide are shown* The
concentrations of these drugs for study are presented.
5. The technique in testing the isolated segments and
strips of Ascaris by the kymograph method are shown*
6. Types of contraction and relaxing of the segments and
strips are described and the effects of different
concentrations of Pentacresol and Arsenamide are
measured.
7. A resume of the artificial media used in i^ vitro
studies by several workers on Ascaris is given.
8. An analysis of the different modes of penetration of
an the Iminthi c s is presented.
LITERATURE CITED
Baldwin, E.
1943 An in vitro method for the chemotherapeutic
investigation of anthelminthie potential*
Parasitol* 89-111.
Baldwin, E.
1948 A study of anthelminthie potency in relation
to chemical constitution. Brit. Jour.
Pharraocol. 91-107.
Baldwin, E., and Moyle, V".
1947 An isolated nerve-muscle preparation from
Ascaris lumbricoides var. suum. Jour. Exp.
Biol. 23, 277-290.
Clark, A.J.
1937 Experimentelle Pharmakologie. Edwards Bros.,
Inc. Ann Arbor, Michigan. 288 pps.
da Costa, G.F.S.
1931 Etude Pharmacologique et thérapeutique sur
l’action anthe Imi nthi que de quelques composes
organiques arsenicaux. Archives int.
Pharmacodym. 443-459.
Hall, M.C., and Poster, W.D.
1917 Efficiency of some anthelminthlcs. Jour.
Agric. Research. 12, 397-423.
Hobson, A.D.
1948 The physiology and cultivation in artificial
media of nematodes parasitic in the
alimentary tract of animal. Parasitol. 58,
183-225.
Lamson, P.D., Molloy, D.M., and Brown, H.W.
1932 Pield studies of anthelminthie action, jbner.
Jour. Hyg. 21, 188-199.
Lamson, P.D., and Ward, G.B.
1936 Earthworms as test objects for determination
value of drugs to be used in human
intestinal helminthic infestations. Sci. 84,
293-294.
27
Oettingen, von, W.P*
1948 Phenol and its derivatives: the relation
between their chemical constitution and their
effect on the organism. U.S. Public Health
Bull. No. 190. 408 pps.
Rebello, Silvio, Rico, T., and da Costa, G.P.S.
1929 La Reactivite des helminthes etudiee par la
methods graphique. C.R. of Soc. Biol. Paris.,
94, 915-923.
Rogers, W.P., and Lazarus, Marian
1948 The uptake of radioactive phosphorus from
host tissues and fluids by nematode parasites.
Parasitol. 39, 245-250.
Stollman, T.
1918 Anthelminthlcs and their efficiency as tested
on earthworms. Jour, of Pharmocol. and Exp.
Therapeutic. 1^, 129-169.
Trendelenburg, Paul
1915 Uber die Wirkung des santonins und seiner
Derivitate auf die Wurmmuskulatur und
Bemerkungen zur Wirkung des Oleum chenopdii.
Arch, fur Exp. Path, und Pharra. 79, 190-217.
Trim, A.R.
1944 Experiments on the mode of action of
hexylresorcinol as an anthelminthie.
Parasitol. 35, 209-219.
PLATE I
28
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PLATE VI
33
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Asset Metadata

Creator Nicholson, Edward Lalonde (author)

Core Title The effects of arsenamide and pentacresol and anthelminthics on Ascaris lumbricoides var. suum

Contributor Digitized by ProQuest (provenance)

Degree Master of Science

Publisher University of Southern California (original), University of Southern California. Libraries (digital)

Tag Biological Sciences,OAI-PMH Harvest

Format application/pdf (imt)

Language English

Permanent Link (DOI) https://doi.org/10.25549/usctheses-c39-276869

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Legacy Identifier EP67195.pdf

Dmrecord 276869

Document Type Thesis

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University of Southern California Dissertations and Theses