A CRITICISM OF COL. LAWRIE'S

EXPERIMENTS.

By C. J. FEAIINSIDE, M.B., C.M., CaPT., I.M.S.,

Superintendent. Central Jail, Jlajahmundri.

The conclusions of Lieutenant-Colonel Lawrie's

paper may be summarized as follows: ?

1. " Laveran's bodies, avian and human, are not parasites.

2. Laveran's bodies are a product of the blood producing no deterioration in health.

3. Our discovery of the rosette in crow's blood proves that MacCallutn's assumption at

all events is false, since it is evident that even if the proteosoma were a parasite, it could not be produced in two entirely different ways. On the other hand, the rosette is proved not to be a sporulating body in bird's blood, by the fact that its spores are a great deal larger than the speck in the red cell, which is the form in which the proteosoma first appears in the blood."

Lieutenant-Colonel Lawrie is of opinion that halteridia and proteosomata are one and the

same, the only difference between them being that the halteridium is halter-shaped, and the proteosoma is round; and when free in the

blood, i.e., not contained in a red corpuscle, they cannot be distinguished from one another. In fact, the shape is the result of mechanical

conditions, namely, the elasticity of the wall of the red cell. I do not wish to argue whether there is an}r difference between the halteridium and proteosoma as regards their being separate parasites, but intend for my pur- pose to accept Lawrie's contention that they are the same in the present instance. The researches at Hyderabad have shown that

the halteridium and the proteosoma are one and the same; and that on the removal of the mechanical restraint of the corpuscular wall the halteridium assumes a circular shape. LabW has previously pointed out that spores are form- ed in two rosettes at the club-shaped ends of the halteridium, the pigment being deposited close by. If the elasticity of the red corpuscle is such as to allow the parasite to assume the spherical shape, we have the spores located in the periphery as a rosette, and the pigment more or less de- posited in the central area. It is evident, there- fore, that the spores are not produced in two

different ways, but rather that their distribution is the result of altered mechanical conditions. The discovery of the rosette by Labbe in pro-

teosoma dates further back than that found in

Hyderabad in 1899. I may quote the following from Le Dantee's work on the sporozoa:?

" Le sans; de l'aloutte renferme un autre ? ? ?/

parasite dont le C3'de evolutif est different du precedent (halteridium). An d^but il n'y a

ancune difference avec halteridium pendant le stade d'accroissement, le corps peut presenter deux formes, uniforme de poire on une forme

THE INDIAN MEDICAL GAZETTE. [Jan. 1900.

amibo'ide: Mais dans tons les cas, au moment

011 la sporulation va commencer, le corps est

spherique: il se transforme tout en tier en

une spore nue avec un nombre variable de

sporozoittes." A second argument is put forward that the

rosette cannot be a sporulating body in bird's

blood, by the fact that its spores are a great deal larger than the speck in the red cell. No measurements are given of the bodies found in the red cells nor of the spores. It must be re- membered that the spore after entering the red corpuscle is, for the same mechanical reasons

previously alluded to, in different surroundings. It looks smaller not only on this account but also 011 account of the wall of the red cell, and a fine layer of haemoglobin intervening.

It is stated that even if the proteosoma were a parasite, it could not be produced in two en-

tirely different ways. By this I understand that dimorphic evolution is impossible, and that two kinds of spores cannot be formed. I have shown that the rosette of the proteosoma and the dual rosette of halteridium are one and the

same, and the altered appearance is only the result of mechanical conditions (always assuming that they are one parasite). Dimorphism has been proved amongst the

coccidia allies of the gymnosporidia, bath be-

longing to the order of the sporozoa. By the application of this dimorphic evolution to the gymnosporidia, Ross, by his brilliant researches, has been able to demonstrate the extra corporeal phase of the malarial hsemamoeba of birds.

In the first place we shall take a look at the spore formation of the coccidia, where we shall find two kinds of spores produced in the same animal simultaneously. For my purpose I shall state briefly the evolution of the coccidiuni salamandrae. This parasite is usually found in the intestinal epithelium, and has two methods of reproduction : (1) endogenous for reproduc- tion in the individual itself; (2) exogenous in- tended for reproduction outside the animal's body, thereby securing the continuation of the species.

Endogenous. Phase (1). The young coccidium in this stage is a small

round body contained in an epithelial cell of the intestine. Inside the sphere is a circular nucleole, which divides into two (kariokinesis) and division continues, but is limited to about 50. Round each nucleole the protoplasm collects and finally separates, each forming a distinct body with a chromatic nucleole inside. The bodies called merozoites now become oval and

finally.crescentic (the crescents shape prevalent amongst the sporozoa is indeed remarkable). The crescents enter fresh epithelial cells and

develop. (Vide fig. 1.) Phase (2)?In phase 1 I mentioned that nu-

clear division was limited to 50 or so, but in a

number of young coccidia the division is not

limited, and in consequence of this the develop- ment is different. A dark central mass is formed to the circumference, of which is attached a number of spindle-shaped bodies, which are pseudo flagellates and stain readily. They are called chroinatozoids. Vide fig. II (male element).

Exogenous cycle.? One of these enter a young coccidium, and what is the result. It attaches itself to the nucleole, partly surrounds it, and finally fusion of the two bodies takes

place. Subsequently throughout the protoplasm a considerable number of chromatic granules appear which were formerly absent. The

coccidium, still deprived of a membrane, increases in size, and the chromatic granules ar- range themselves peripherally, unite and form a membrane, in the interior, of which is a

granular mass of sporoblastic protoplasm. The

coccidium, now protected by a capsule, passes out or the intestine, and under favourable conditions four spores ave formed, which give rise to eight sporozoites. The sporozoite having entered the intestine of another salamander, the C3rcles begin over again. (Vide fig. III.)

For the sake of argumemt I have assumed Colonel Lawrie is correct in his view that pro- teosoma Labbe and halteridia are the same, and in having done so I have shown that the forma- tion of the spores is the same in both, and that mechanical conditions cause the rosette in the proteosoma and the dual rosette in the halteridia. This is the endogenous evolution which has its analogue amongst the coccidia. Phase 1 is represented bj' the deposition of spores in the blood (inerozoite). Phase 2 by the develop- ment of the flagellate (chromatozoid). The

exogenous cycle, worked out by Ross, is the union of a flagellate with a proteosoma which give rise to zygotoblasts (sporozoite) found in the salivary gland of infected culex pipiens. Take the malarial parasite of man as another

example, say the spring tertian. It deposits spores in the blood (merozoites) ; it throws out

flagellates (chroinatozoids); it forms zygotes in the stomach wall of anopheles by the union of a flagellate (chromatozoid) with a (female) hajmamceba. Ross has shown that in the case

of proteosoma zygotoblasts there are two kinds of sporozoites, the one thread-like and the other a small brown bod}\ The former is supposed to cause infection by inoculation, the latter by water or some other medium. Did the Hydera- bad observers exclude the possibility of infec- tion amongst the birds experimented on ?

MacCallum has proved that proteosoma are composed of two elements, a hyaline cell which throws out flagella (male element), and the other a granular cell (female). I have myself seen the conjugation of the two taking place, and I

have no doubt that impregnation must have taken place. Bignami and Bastianelli describe minutely the mode of development outside the human body of the spring tertian parasites*

Jan. 1900.] THE INDIAN MEDICAL GAZETTE.

EVOLUTION OF COCCIDIUM SALAMAMDL?JE.

ST\

( Halteridium and Evolution of

proteosoma of Birds.

Fig (IV). Halteridium Endogenous Cycle

Phase (1).

O) (N r

^6*1 /r, ;

tsS h>-.443' ? 2 |

^ O -O 1

H.. Pk

(i) a (2)

EVOLUTION OF COCCIDIUM SALAMAMDL?JE.

Phase (I).

Fig- (I).

Phase (2).

Fig. (II).

Fig. (III).

Fig (IV). Halterulium Endogenous Cycle.

Phase (1).

Fig.

(V).

Endo

geno

us Cycle,

P. Labbe ?Phase (

2).

Fig.

VI.

Exogenous Cycle

of

P. Lab

be.

.Tax. 1900.] LIE17T.-COT.ONEL LAWRIE'S REPLY.

Tliey are of two kinds, a smaller micro-gamete (male element, cliromatozoid), and a larger, the micro-gamete, containing the female element. In the September number of this journal, I de- scribed certain spring tertian parasites as break- ing up into two or more small pigmented spherules, each carrying pigment in active motion and containing chromatin. Other spheres became flagellates, and I suggested that tli3

, CO

former was the female element and the latter the male (spermatozoid).

In conclusion an endeavour has been made to

prove that?

(1) The rosette of the Hyderabad observers was anticipated by Labbe himself.

(2) Assuming proteosoma and halteridium to be the same, the arrangement of the spores is due to mechanical conditions similar to those ex-

planatory of the shape of the parasite. (3) The presence of the rosette in the pro-

teosoma is only the endogenous cycle of the parasite, a fact which tends to confirm rather than detract from Ross' discovery of the exoge- nous cycle. I am indebted to my jailor, Mr. Mit- chell, for the illustrations. Figs. I, 11, III being taken from a paper on work done at the Pasteur Institute. Fig. IV is taken from Labbe's illus-

trations. Fig. V represents the exogenous evo- 11

lution of proteosoma Labbe in the stomach wall of culex pipiens, and is taken from 1113' notes on

infected moscjuitoe.s. PLATE.

Fig. I.?Endogenous cycle, phase (1). (a, b. c). Young coccidium in epithelial cell (kar-

zokinesis). (d). Division is limited. (e, f) Development of merozoites, (f) being a

meroi.oite which enters a fresh epithelial cell of the host.

Fig. II.?Endogenous cycle, phase (2), (a, b, e). Young coccidium, where nuclear division

is not limited.

(//, e). Formation of chromatozoids. (J). Chromatozoids.

Fig. HI.?Exogenous cycle. (a, b). Cliromatozoid has entered young coccidium and locates itself near the central nucleole.

(c). Fusion of chromatozoid with nucleole. (d,j). Formation of spure {d, e, f) inside the ani-

mal ((], h, i,j, ft) further development after it

has left the animal.

(/i). Spore (1), sporozoite (2) two in number in each spore.

Fig. IV. ?Endogenous cycle of halteridium with rosettes (?, j). (Libbe).

Fig, V.?Endogenous cycle of proteosoma phase (2) union of a chromatozoii with a young proteoscme.

Fig. VI. ?Exogenous cycle of proteosoma in the stomach wall of culex pipiens infected experimental- ly by me.

(a). Young zygote after impregnation. (b). Achrepseudopodial movements taken on to en-

able it to move into the stomach wall.

(r-). Vacuolatiou and linear striation. (d,e,f). Zygoloblasts forming from the cytoplasm (ff). Oyst found in the intestine (an unusual situa-

tion) (1) low power; (2) highly magnified. Linear, zygotoblasts and brown spores are visible

Remarks on above paper by Dr. Lawrie. Labbe discovered that "spores" are formed,

"in two rosettes, at the club-shaped ends of the halteridium." We do not pretend to have either discovered or observed this phenomenon; and I am not aware of any description which corre-

sponds with our drawing,from life, of " the rosette

in the elliptical blood cell of a crow," in the

November issue of this journal. In the second place it was considered sufficient

to state that the speck, the first visible com-

mencement of the halteridium or proteosoma, in the red corpuscle is actually much smaller than the so-called spore of the rosette. The difference in size is so apparent that measurement is unne-

cessary. Moreover the objection that the speck in the red corpuscle looks smaller than it really is? which we do not admit to be the case?applies equally to all other forms of the Laveran body which are produced in its interior. It is a well

known fact, however, that neither the " wall

" of

the red corpuscle, nor its contained haemoglobin, has any appreciable optical effect on the intra-

corpuscular forms of the Laveran body. In the third place Dr. Fearnside understands

us to mean that "dimorphic evolution is im-

possible, and that two kinds of spores cannot be formed." This is not our meaning. What we mean is that even if the proteosoma were proved to be an animal parasite, it could not

reproduce itself on the one hand like the coccidia, which are among the very lowest types of animal life, and on the other hand by a sexual process identical with that which obtains in the

highest animals. But we go much further than this. We deny altogether that either the

halteridium or the proteosoma, or any form of the Laveran body, avian or human, possesses the function of reproduction at all. The rosette is met with so very rarely in the blood either of birds or of human beings that it is impossible to regard it as the sporulating element by which reproduction is carried on, and when it is found, the proteosomata forthwith dimin- ish instead of increasing in number. What Dr. Fearnside asserts regarding the application of dimorphic evolution to gymnosporidia only confirms our view that Ross' theory of malaria is founded entirely on assumption and not on fact.

MacCallum has proved nothing He has mad? statements which have been accepted by Manson and Ross, and made use of by them without having been verified. MacCallum's momentous contri- bution to the parasitology of malaria?on which the whole mosquito theory now hangs?was a sexual process among the Laveran bodies of birds, which results in an offspring with a beak which destroys the blood corpuscles wholesale ; but no such Laveran body exists. The body with a beak which MacCallum describes was most

likely a monad, which is sometimes met with in

THE INDIAN MEDICAL GAZETTE. [Jan. 1900.

birds' blood, and is the only thing which in any way tallies with his description. Dr. Fearnside

ought to be aware that such a breaking up as he describes, of " certain spring tertian parasites into two or more small pigmented spherules," takes

place only when the blood itself breaks up, and is obviously due to mechanical causes such as the pressure of the cover-slip, etc. Finally, there is no more ground for supposing that the

flagellum is a spermatozoid than there is for

supposing that pseudopods are sperinatozoids. The points in controversy between ourselves

and the plasmodists cannot be settled by mere assertion and counter-assertion. They should be dealt with b}r a Commission or Committee of impartial observers, of whom there are many in India who are fully competent to note the facts and draw correct conclusions from them. I am aware that our researches appear to go in the very teeth of Manson, Ross, Koch, and other well-known authorities on the malaria " para- site." Truth, however, is not the child of

authority but of time.