THE ACTION OF DRUGS ON DYSENTERY BACTERIOPHAGE *

By G. PANJA, m.b. (Cal.), D.Bact. (Lond.)

Officiating Professor of Bacteriology and Pathology, School of Tropical Medicine, Calcutta

(From the Department of Bacteriology and Pathology, School of Tropical Medicine, Calcutta)

Questions are often asked by practitioners whether dysentery bacteriophage and chemo-

therapeutic drugs could be given simultaneously and whether the action of phage is hindered by these drugs. The following experiments were

undertaken in answer to such questions..

First experiment A strong dysentery phage was diluted by mix-

ing 1 drop with 20 c.cm. of broth and from this

mixture 5 drops were added to 100 c.cra. of broth. A small quantity from this final dilution was mixed with an equal quantity of young broth culture of Bact. jlexneri and plated on pepsin agar medium. Discrete phage plaques were seen after 18 to 20 hours' incubation. The above diluted phage as well as strong phage were used for the experiments. Solutions and suspensions of the drugs were made in distilled water and kept in contact with equal quantities of phage at room temperature for one to two hours and then filtered through L3 candles. The filtrates were tested for the presence of phage, and the results are given in table I.

Phage could be recovered from the area of lysis. Distinct plagues indicated that lysis was due to phage action and not due to the bacteri- cidal action of the drugs, as bacteria were found not killed by the drugs outside the plaques.

It will be seen that the above three chemo-

therapeutic drugs in 1 per cent strength, phenol 0.5 per cent and electrolytic chlorine 0.25 p?r cent do not inactivate dysentery phage in 1 to

2 hours in vitro. Phage boiled for 2 minutes is inactivated.

Second experiment

This was done with double the concentrations of solutions and suspensions, and an increase in the time of contact. For results see table li-

lt will be seen that 2 per cent strengths of the above chemotherapeutic drugs and phenol 1 Per cent do not inactivate the dysentery phage in 2 to 3 hours in vitro. Electrolytic chlorine 0.25 per cent partially inactivates it in 3 hours-

Boiling for 5 minutes kills the phage. Clear areas are due to lysis of bacteria, as the drugs alone do not give rise to such clear areas.

Table I

Growth of dysentery phage in solutions of the sulpha drugs, etc.

Drugs

Percentage

Time of contact, hours

A loopful of filtrate touched on culture of

dysentery bacilli spread on solid medium.

A loopful mixed with 3 to 4 loopfuls of cul- ture and spread.

1 to 2 c.cm. of filtrate mixed with 10 to 15

drops of culture and

spread.

M.&B. 693

1

1

L

+

++

Sulpha- guanidine

1

1

L

H?b

++

Solu- septasine

1

2

L

+

Phenol

0.5

2

L

+

chlorine 2 minutes any

0.25

2

L

+ +

+

L ?complete lysis; + = plaques present; =no plaques, no lysis.

Control, .e. phage without any

treatment

0

L

+

+++'

* The paper was read before the Indian Science Congress Association held at Nagpur in January 1945.

June, 1945] ACTION OF DRUGS ON DYSENTERY BACTERIOPHAGE : PANJA 295

Table II

Growth of dysentery phage in solutions of the sulpha (bugs, etc., in increased concent)ations and time of contact

Drugs M.&B. 693 ifee

Percentage Time of contact, hours A- loopful of filtrate touched on spread culture of dysentery bacilli.

One loopful of filtrate mixed with one loop- ful of culture and spread.

Equal amounts of filtrate mixed with equal amounts of cul- ture in liquid medium. A loopful of drug alone touched on spread culture.

+++ +++ ++++

'

Electrolytic Plieno1 1 chlorine

0.5

3

(L)

Phage Control boiled for without 5 minutes boiling

+++ ++ - +++

(L)

(+) (+)

(+)= Partial clearing. (L) = incomplete lysis.

Table III

Growth of undiluted dysentery phage in solutions of the sulpha drugs, etc., in increased _ concentrations and time of contact

Drugs | M.&B. 693

^ercentage ? ? j ^

Time of contact, hours j 2

Sulpha- ; Solu- guanidine ; septasine

,e loopful of filtrate ouched on spread

bac^f8 dysentery

drop of filtrate JXed with 5 to 6 c.c. broth culture and

sPread.

L

+n

2

24

L

+

I n n

+

Phenol

1

24

L

+n

Electrolytic chlorine

0.5

24

L

Boiled for 5 minutes

4

(L)

Control

+n t

4

L

+n

+n = confluent phage plaques and no bacterial growth; (L) ineomplott hsis, (+)

colonies are present.

Third experiment icith undiluted strong phage

, One drop of strong phage is added to the rugs. The time of contact and concentrations

are as in table III.

Conclusion ,, Two per cent strengths of the above chemo- lerapeutic drugs, phenol 1 per cent ana

electrolytic chlorine 0.5 per cent have no ac-

tion on strong dysentery phage. Boiling of a strong phage for 5 minutes does not com-

pletely kill it. It was not considered necessary to carry on

the experiments further, as in practice, the above chemotherapeutic drugs and the bacteriophage do not reach such a high concentration in the gut.