Effects of various chemicals on Paramecium caudatum

N O T E S F R O M T H E BIOCHEMICAL RESEARCH F O U N D A T I O N . Effects of V a r i o u s C h e m i c a l s o n Paramecium caudatum.--Hucit E. POTTS. T h e effects on paramecium of a series of organic arsenicals (JOURNAL OF THE FRANKLIN INSTITUTE,236 : 499, 1943) and of propionic acid and its sodium salt (JOURNAL OF THE FRANKLIN INSTITUTE, 236: 99, I943) have been reported. T h e present paper deals with the effects on paramecium of various chemicals which do not constitute a single group of related compounds. Several of the c o m p o u n d s were synthesized by members of the Laboratory staff, Nos. 4 and 5 as new syntheses (Journal of the American Chemical Society, 62: 3508, I94O) and Nos. I, 2, 3, 20, 2I, 22, 23, by standard procedures. T h e m e t h o d used in testing the substances was the same as t h a t previously described with the exception t h a t the n u m b e r of paramecia was kept as nearly constant as possible. Eight drops of paramecium culture were placed in each of five watch glasses, the fifth being used as a control. To these, 8, 4, 2, I and o drops, respectively, of a given drug solution were added. T h e total n u m b e r of drops in each glass was made up to I6, by adding a sufficient n u m b e r of drops of filtered, boiled and cooled water taken from cultures of paramecium. Each c o m p o u n d was neutralized to approximately pH 7.o prior to testing. T h e results are shown in Table I. Benzenestibonic acid appeared to be more active t h a n the pentavalent benzene arsenicals (JOURNALO F T H E FRANKLININSTITUTE,236: 499, !943). T a r t a r emetic showed about the same order of activity as benzenestibonic acid. Sulfanilamide was conspicuously ineffective. It might be t h a t this type of drug would have some effect on the ability of paramecia to reproduce, b u t this has not been tested as yet, since inhibition of reproduction in cultures with bacteria present would not indicate whether the action was on paramecia or on their food supply. T h e action of quinine plus urea HC1 U.S.P. XI, which killed paramecium in I : 25,000, is fairly close to t h a t of quinine alone on paramecium as shown by Feller (Archly fCir Protistenkunde, 59:562, 1927) where I : IOO,OOO was fatal. T h e action of sulfanila~nide on paramecium, killing in 1 : 200, is of the same general order as shown by Ferguson et al. (Journal of the Elisha Mitchell Scientific Society, 58: 53, I942 ) on certain free-living protozoa. Phenylmercuric acetate and phenylmercuric nitrate were exceedingly active against paramecia, as were also 8-hydroxyquinoline, diethyl227

228

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TABLE

FOUNDATION.

[J. F. I.

I.

Effect of Various Chemicals on Paramecia. No. of '.hemical.

N a m e of C h e m i c a l .

Concentrations Tested.

E f f e c t on Paramecium caudatum.

I

3 - P y r i d i n e s u l f o n i c acid

I :

I26 I : 629

Killed all in 3 ° min. Killed m a n y in 30 min. b u t failed to kill all in 24 hrs.

2

Potassium-5-nitro-epyridinesulfonate

:83 i : 413

Killed all in 30 min. Killed m a n y b u t n o t all in 24 hrs.

3

5-Amino-2-pyridinesulfonic acid

I :

330 I : 575

Killed all in 24 hrs. Failed to kill a n y in 24 hrs.

4

5-Nitro-2-pyridyl phenyl sulfone

s a t ' d soln. (less t h a n

Killed all in 3 hrs.

I : I,OOO)

5

5-Amino-2-pyridyl phenyl sulfone

s a t ' d soln. (less t h a n

Failed to kill a n y in 24 hrs.

I : I,OOO)

6

B e n z e n e s t i b o n i c acid

< I : 743 929

< I :

Killed all in 24 hrs. Killed a few b u t n o t all in 24 hrs.

7

Berberine

I : IO,OOO I : 20,000 I : 40,000

Killed all in 2 hrs. Killed all in 24 hrs. Failed to kill a n y in 24 hrs.

8

8-Hydroxyquinoline

I : I~OOO,OOO I : 2,000,000

Killed all in I to 2½ hrs. (2 tests) Killed all in I hr. (2 tests) (no g r e a t e r dilution tested)

9

Phenylmercuric nitrate

I : 1,600,000 I : 3,200,000

Killed all in 24 hrs. Killed a few b u t n o t all in 24 hrs.

IO

Phenylmercuric acetate

I : 1,600,000 I : 3,200,000

Killed all in 3 hrs. Killed all in 24 hrs. (no g r e a t e r d i l u t i tested)

II

Tartar emetic

I :

I : 4o0 800

Killed all in 5 to 24 hrs. (2 tests) Killed m o r e t h a n ½ b u t n o t all in 24 h

12

Sulfanilamide

I : 200 I : 400

Killed all in 24 hrs. Killed m o r e t h a n ½ b u t n o t all in 24 h

13

Acridone

I : 500

Failed to kill a n y in 24 hrs.

I4

Atebrine

i : 51,2oo I : IO2,4oo I : 204,800

Killed all in 6 hrs. Killed all in 24 hrs. Killed a few b u t n o t all in 24 hrs.

I5

Rhodamine B

I : 2o,0oo I : 4o,ooo

Killed all in 6 hrs. Killed a few b u t n o t all in 24 hrs.

x6

Rhodamine G.D.N.

I :

I6O,OOO i : 320,000

Killed all in 24 hrs. Killed m o s t b u t n o t all in 24 hrs.

z7

Fluorescein

I : 2oo

Killed m o s t b u t n o t all in 24 hrs.

i8

Q u i n i n e + u r e a H C I U.S.P. XI

I : I :

I2,8oo 25,600

Killed all in 6 hrs. Killed m o s t b u t n o t all in 24 hrs.

Mar., 1944.1

BIOCHEMICAL

RESEARCH

FOUNDATION.

22 9

"FABLE [ (Cont'd). No. of Chemical.

Name of Chemical.

Concentrations Tested.

Effect on Paramecium caudatum.

Killed all in 6 to 24 hrs. (2 tests) Killed all in 24 hrs. Failed to kill a n y in 24 hrs. (solution had been left standing at room t e m p e r a t u r e for several days)

19

C o n t r a m i n e (B.D.H.) (Diethylammonium diethyldithiocarbamate)

I : 25,600 I : 5Is2OO I : 25,6o0

20

Diethylammonium benzyldithiocarbamate

I : 1,024,ooo Killed all in 24 hrs. I : 2,048,000 Killed most b u t not all in 24 hrs.

2I

A m m o n i u m phenyldithiocarbamate

I : 16,oo0

Killed all in 3 hrs.

2 IA

A m m o n i u m phenyldithiocarbamate

I : 32,000 I : 64,000

Killed all in 4 hrs. Killed all in 4 hrs. Note: This c o m p o u n d was left for 24 hrs. in refrigerator prior to 2nd test, a n d appeared to be in process of decomposition.

22

Benzylammonium benzyldithiocarbamate

I :

23

p-Chlorophenylammonium p-chlorophenyldithiocarbamate

I : 256,O00

512,OOO Killed all in 24 hrs. I : 1,024,OOG Killed a few b u t not all in 24 hrs. Killed only a very few in 24 hrs. (no greater concentration tested)

ammonium benzyldithiocarbamate and benzylammonium benzyldithiocarbamate. A noticeable weakness in the use of an organism such as paramecium to detect substances which might be of therapeutic value in the treatment of protozoal or other diseases is that such substances as sulfanilamide would be entirely overlooked should the drugs be chosen only on the basis of killing action in great dilution. It is planned to culture paramecia bacteria-free and test for inhibition of reproduction as well as killing action of the drugs, thus taking into consideration two types of activity shown by drugs. Summary and Conclusion: A number of related and some unrelated compounds were tested for their effects on paramecium. The most active were phenylmercuric acetate, phenylmercuric nitrate, 8-hydroxyquinoline, diethylammonium benzyldithiocarbamate and benzylammonium benzyldithiocarbamate. Sulfanilamide was one of the most inactive substances. Of the recognized drugs tested it is seen that quinine, atebrine, 8-hydroxyquinoline, contramine and possibly berberine would have been singled out for possible therapeutic use on the basis of their high activity. The germicides, phenylmercuric nitrate and phenylmercuric acetate, would also have been singled out. Such an effective drug, however, as sulfanilamide would have been entirely disregarded. It is possible that

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[J. F. I.

sulfanilamide and drugs that have similar action may have an inhibitory action on reproduction of protozoa as they do on bacteria. Plans have been made to test such drugs on bacteria-free paramecia in order that the effects may be judged both on the basis of killing action and on inhibition of reproduction.

Abstract of The Adaptation of Magnetic Support to Air-Driven Ultracentrifuges.--THoMAS J. DIETZ AND TERREL V. KISHBAUGH (JOURNAL OF THE FRANKLIN iNSTITUTE,236: 445, 1943). Air-driven ultracentrifuges usually employ air support for the rotor, while those driven electrically use magnetic support. By adapting the magnetic support to the air-driven pattern, it is possible to secure the best features of each type while avoiding some of the disadvantages inherent in each. The use of air drive obviates the necessity of cooling the driving motor, while the magnetic support avoids the vibration sometimes found in the air-supported type. T h e r e is an additional advantage in favor of the magnetic support in that the stray flux provides a magnetic field for the speed indicating device. Furthermore, because of inherent constructional features, the magnetic support is superior, since there is less danger of breakage of the shaft in case of support failure. The magnetic support consists essentially of a solenoid with its armature mounted on the main shaft of the centrifuge. The armature is contained between the two bearings. The current in the solenoid may be adjusted to support nearly all the weight of the rotor, leaving a slight residual thrust against the lower bearing. An alternate method increases the solenoid current until the rotating system is lifted bodily and held with a slight force against the upper bearing. Abstract of A Recording and Controlling System for Air-Driven Ultracentrifuges.--THOMAS J. DIETZ (JOURNAL OF THE FRANKLIN INTITUTE, 236:451, 1943). The operation of ultracentrifuges is considerably simplified by the addition of speed recording and controlling equipment. It is not generally realized, however, that such devices can be readily assembled from standard parts commercially available. An installation at the Biochemical Research Foundation, constructed in this way, offers m a n y new features. The principle of the method is one in common use, whereby the rotational speed is measured by determining the frequency of an alternating current generated in a pick-up coil by a single pole armature, attached to the rotating shaft and magnetized by stray flux from the supporting solenoid. The output of the pick-up coil is fed to an electronic frequency meter made by the General Radio Co. This provides continuous indication of the speed on its main dial, and also supplies actuating current for the Bristol Pyromaster which is the heart of the recording and controlling system.