1 A REPORTON THE MICROSCOPIC OBJECTS FOUND IN CHOLERA EVACUATIONS, &c. }SY TIMOTHY RICHARDS LEWIS, M.8., ASSISTANT BURGEON, HEB MAJESTY'S BEITIBH FORCBS ATTACHED TO THE SANITARY COMMISSIONER WITH TIIH GOVT. ()!<' INDIA.. |)rinteb bj) otim of (Dobnitnmtt. CALCUTTA : OFFICE OF SUPERINTENDENT OF GOVERNMENT PRINTING. 1870. LIBRARY OF THE PRUDENTIAL INS. CO. OF AMERICA N EWARK, N. J. STATISTICIAN'S DEPARTMENT Section JjyiLH . . ... __ Su bject Date%ecd Acknowledged Indexed C 0 4 14 PREFACE. In accordance with instructions issued at the commencement of this inquiry, attention lias been specially directed towards obtaining Tacts bearing' on the truth or otherwise of two hypotheses regarding the cause of cholera — namely, the theory of its fungoid origin, particularly the one advanced by Professor Hallier of J ena ; and the theory of the connection existing between cholera and certain conditions of the soil, promulgated by Professor Max yon Pettenkofer of Munich. In both theories the existence of a specific poison of an organised nature is maintained — a germ ; and both savants belie v o it to exist in the alvine discharges of a person affected with cholera. The Munich Professor does not risk an opinion as to whether it belongs to the animal or to the vegetable kingdom, but infers that the soil is the nidus in which it grows ; whereas Professor Hallier maintains that it multiplies in the human body, and unhesitatingly affirms it to be a fungus. An account of the observations which have been made in order to test the views ad\ r anccd by Professor Hallier will occupy the first portion of the report ; and, as in the course of the investigation my attention has been directed to a consideration of the microscopic objects which are found in the evacuations of cholera patients, a description of them will at the same time be given ; together with illustrations of various initiatory experiments bearing on the general question of ' disease-germs.' T. 11, L. April 1870 TABLE OF CONTENTS. PART I. CONCERNING THE THEORY OP THE FUNGOID ORIGIN OF CHOLERA AND THE MICROSCOPIC OBJECTS FOUND IN CHOLERAIC EVACUATIONS. Epitome of Professor Hallier's published views regarding the existence of a cholera-fungus. Its cyst, its spore, and its micrococcus ? , ? , 1-4 Section 1. The bodies found in choleraic evacuations which may be said to bear some resemblance to cysts ... ... ... ... ... 4-17 The cholera bodies of Drs. Budd, Brittan, and Swayne ... ... 5 The effect produced by various re-agents upon these cyst-like bodies ... 7 The nature of these bodies ... ... ... ... ?, 9 The various methods adopted during the course of the .investigation with the view of ascertaining whether other cyst-like bodies could be artificially developed in cholera stools. Illustration I. — Three cultivations of cholera evacuation in various media ... 10 Illustration II. — Cultivation experiments with the contents of the small intestine of a person who had died of cholera ?, ... Mt 11 Description of the Isolating Apparatus used in some of the experiments ... 12 Illustration lll— Three cultivations of choleraic discharge, two of the samples having been placed upon slices of fruit ... ... « ... 12 Illustration IV.— Cultivation of a choleraic discharge obtained from a locality where the disease was epidemic ... ... 1M 14 Illustration V. — Cultivation of ordinary alvine discharge ... ... 15 Conclusions drawn from the foregoing experiments ?, ... lt , 17 Section 11. The bodies found in choleraic evacautions which may be said to bear some resemblance to spores. 1. Globules of a fatly nature. Effect of re-agents upon them... ... ... M| ?, 18 Their artificial production ?, ?, ?, 20 6 2. Altered condition of blood-cells. Exact microscopic appearance of such ... ... ... ... 21 The blood-cells observed to protrude and retract portions of their substance 21 Precisely similar appearances observed in " chylous urine" in wliieli the embryos of a worm were discovered ... ... ... ... 22 3. The corpuscles associated with the flocculi in rice-water stools. Illustration I. — The changes which occurred in the appearance of these corpuscles, which when first seen were hyaline ... ... ... 2:? The effect produced by various re-agents upon them ... ... ... 21. Illustration II. — The same as the foregoing, except that the corpuscles were granular when first examined ... ... ... ... 25 Illustration 111. — The same as the foregoing, the hyaline and granular appearance of the corpuscles being evident in the same preparation ... 20 4. Globular conditions of certain Animalcules. Illustrations I — III. — In which the various stages of these bodies, hitherto observed, are described ... ? , ... ... 20 Are any of these four classes of corpuscles peculiar to cholera ? ... ... 31 Significance to be attached to the presence of the first and second classes of bodies referred to ... ... ... ... ... 31 Significance to be attached to the presence of fourth class of bodies referred to 82 Significance to be attached to the presence of third class of bodies referred to 31 Probable identity of the last named with the " peculiar corpuscles " of Professor Parkes ... ... ... ¦¦• ••¦ ••• 35 Reasons for concluding that they are not, as generally stated, disintegrated epithelial cells ... ... ... ... ... 35 Their probable nature ... ... ... ... ... 30 Sectiok 111. Observations as to the nature of the minute molecules {micrococcus) observable in cholera stools, preceded by illustrative experiments ol' the changes which were seen k> occur in other solutions of organic matter ... ... 37 Illustration I. — The changes which were observed to occur in solutions of organic matter (meat) under three diiferent conditions ... ... 40 Illustration II. — The changes which were observed to occur in filtered solutions of ordinary alvine discharges ... ..'. ... ... 41 Illustration lll.— The changes which were observed to occur in two preparations of an ordinary alvine discharge which had been placed on the same slide ... ... ... ... ••• ••• 46 Various developmental changes observed, from simple molecules to the advent of a " nucleated" infusorium ... ... ... ... 47 Illustration IV. — The changes which were observed to occur in five specimens of choleraic dejecta under varying conditions ... ... ... 51 Lessons to be derived from the experiments illustrating this section ... sli Summary of conclusions drawn from all the experiments ?, ... 54 PART 11. BEMABKB EEGABDING THE SOIL, &C., OF CEBTAIN PLACES IN RELATION HO PETTENKOFEII's THEOEY. PAGE. A summary of Professor Pettenkofer's views concerning the relation existing between the existence of cholera and the condition of the ground-water ... 58 Dr. Buchanan's explanation of this theory ... ... ... 00 Various observations bearing more or less directly on the foregoing, in connection with short topographical descriptions of the following places, with special reference to their relation to water; in addition to which, the extent of the permeability of the soil, the percentage of organic matter in it, and the result of its microscopical examination in a moistened condition are given in detail. The method adopted in order to ascertain the exact extent to which the soil is porous to air and water will be found in connection with the first named station. Allahabad ... ... ... ... ... ... 62 Cawnpore ... ... ... ... ... ... (56 Lucknow ... ... ... ... ... ... 67 Fyzabad ... ... ... ... ... - 70 Agra ... ?. ... ... ... ... 71 Morar and Gwalior ... ?., ... ••• ••• 71 Meerut ... ... ... ... ... ... 71 Peshawar ... ... ... ... ... ... 70 Concluding Remarks ... ... ... ... ... 77 A CONDENSED DESCRIPTION OF THE ILLUSTRATIONS FIGUBES. If OS. Plate I (opposite page 1.) A copy of HalUers drawing of Ike clwler a fungus. Mature cholera " cyst" swollen and ruptured I 2 i. Cholera cysts less mature Swelled "spores," which were supposed to have escaped from cholera cysts; some of them are seeu degenerating into " Mierooocous" 3 " Micrococcus Colonies" — (a) Colony formed by the breaking up of a single spore, (b) Ditto still further broken up. ('') A group of " Colonies " corresponding to several spores, (d) Germinated Micrococcus ... 1 " Micrococcus" in process of germination .. Ditto giving rise to filaments X ;> (J Highly developed filament with cyst (c), and macroconidia (m) A cholera cyst or sporangium still attached to its fertile filament.. Filaments illustrating the tendency to the formation of Tilletia 7 8 caries. What was considered a matured spore of the latter is marked sp ... 0 I) An aggregation of " cholera cysts" A "cholera cyst" germinating.... lo 11 Plate II {opposite page 2.) The cholera bodies of Drs. Budd, Brittan, and Swayne (after liohin) Brittan's "annular bodies" in cholera (copied from Medical Gazette) Swayne's "cholera cells" (copied from Lancet) ii. in. iv Plate 111 (opposite page 4.) Cysts closely resembling Brittan's cholera bodies ; consisting principally of fatty matter enveloped by fibro-albuniinous material ... v.-vn. A globular cyst-like body observed in choleraic dejecta Effect of liq. potassse upon No. 1 v. I 2-4 Two sizes of the globular cyst-like bodies as at v. 1 Appearance after the addition of acetic acid 1-2 VI. 3-5 Globular cyst-like body surrounded by a compact fibro-albuminous layer VII. Effect of ether after the previous application of liq. potassse Bodies resembling the "cholera cells" of Swayne. They are ova of ordinary Hound- worms DESCKiriMON OF ILLUSTIi A'l lONS. 10 FIOITBKS. NOS. Ova, as commonly met with in alvine discharges ... ?. viii. 1-1 Ova,, the contents having assumed a somewhat dolined arrangements ? 5-6 Embryo completed „• ••¦ ••¦ ••• •¦• viii. 7 Embryo escaped .. ... ••• ... ... ? 8 Same as viii., the form having been altered by pressure ... ix. Effect of adding ether ... ... ... ... „ 1 Aspect assumed after the addition of liq. potnssre subsequent to the application of ether ... ... ... ... „ 2-5 Same as viii. — Also treated with ether. No. 3 was ruptured by pressure ... ... ... ... ... x. 1-5 Plate IV (opposite page 6.) More highly magnified specimen of Fig. viii ... ... xi. After the addition of acetic acid ... ... ... ? 1-2 . of iodine and absolute alcohol ... ... ? 3 . of absolute alcohol 0n1y... ... ... „ 4 Ova of acarus (domesticus ?) — sometimes found in choleraic and other dejections ... ... ... ... xii. Partly disintegrated acarus obtained in a cholera stool (magnified by a low power) ... ... ... ••• xiii; Highly stained specimen of the ovum of Tricoceplialus (diapar ?) — " probably the body delineated at No. 2, Fig. iii, in Dr. Brittan's Ditto ruptured by pressure ... ... ... ... „ Highly stained specimen of the ovum of an ascaris found in the H same stool (cholera) as the foregoing,— ruptured by pressure ... „ 3 Mycelium escaping from an aggregation of molecules (micrococcus). Spores not visible (cholera stool) ... ... ... xv. Germinating spores, together with mycelial filaments (cholera stool) xvi. Plate V (opposite page 8.) Fungus developed in a cholera stool. A later condition observed In the preparation delineated at xv ... ... ... xvii. Spores, some of which have germinated ... ... •• » 1-2 Mycelium, upon which dilatations or macroconidia (m) are seen ... ? '•* Filaments with bulbous terminations ... ... ... „ -I Fertile filament terminated by a cyst or sporangium, the contents of which is seen to have contracted within the capsule ... „ <¦> The "Isolating Apparatus" used in some of the experiments ... xviii. Funnel containing a plug of cotton wool ... ... ... >, 1 Flask containing strong sulphuric acid ... ... ... » Shallow dish (containing a solution of permanganate of potash), with an inverted bell-glass, inside of which is a small wire stage for elevating the preparation above the level of the fluid in the dish „ 3 An Aspirator filled with water. One arrow represents the escape of the latter, and the other arrow shows the course which the entering air has to take before it can replace the escaped water ... •< 4 Plate VI (opposite page 10.) JMugi which were developed in a cholera stool ... ... xix. DESCIUPTION OF ILLUSTRATIONS. 11 FIGURES. NOS. Fertile filament of Aspergillus ; some of the spores (conidia) are seen falling off... ... *¦¦ ••• • ¦• xix. 1 Ditto ditto Penicillium ... ... ... „ 2 Cells of various sizes in the cultivation, probably modified spores ... „ 3 Very thin filaments terminating in excessively delicate mucor-like cysts or sporangia, some of which are filled with elongated spores ... ... ... ••• ••• » 4 Plate VII {opposite page 12.) Highly developed specimens of mycelial filaments, with numerous dilatations {Macroconidia), which separating are found as freo circular cells in the field, capable of germinating like ordinary spores (cultivated in cholera discharge) ... ... xx. Plate VIII {opposite page Li.) „ Fungi developed in ordinary evacuation. Spores in process of germination ?. ... ?. xxi. 1-2 Microcoecus ... ... ... ... ... „ 3 Penicillium glaucum ... ... ... ... xxii. 1 Aspergillus ... ... ... ... ?. „ 2 Numerous filaments of Oidium lactis, corresponding to the " cholerafungus" of Thome ... ... ... ... xxiii. Plate IX {opposite page IG.) Fungi developed in ordinary evacuation. Spores, cysts, and filaments of Mucor in various stages of development ... ... ... ... ... xxiv. Escaped spores ... ... ... ... ?, „ i Detached cyst or sporangium ... ... ... ?, „ 2 Cysts still attached to the fertile filaments ... ... ?, „ 3 Heads of fertile filaments (Columella), with the remains of the ruptured cyst-capsules still attached ... ... • ... „ 4 jy. B. — Compare Nos. 2 and 3 with Hallier's figures (Plate I, Nos. 2 and 8). liuptured mucor sporangia ... ... ? ... xxv. A ruptured cyst with spores escaping ... ... ?, }J 1 Ditto the spores having completely escaped ... ?, „ 2 Ditto detached fiom its stalk ... ... ... „ 3 A mucor cyst detached from the fertile filament. The spores are seen to escape through the capsule ... ... ... xxvi. 1 Aspergillus fructification simulating that of mucor; a glutinous film surrounding it, thus keeping the spores or conidia together. The fertile filament is seen to be partly ruptured ... ... ? 2 Detached Aspergillus heads of various sizes, the spores being held together by means of some glutinous material ... xxvii. 1-2 Ditto in process of germination. N. B. — Compare with Hallier's drawing of the mature cholera-cyst in the same condition (Plate I,No. 11.) ... ?, " ... ... ... „ 3 12 DESCRIPTION 01 ILLUSTRATIONS. FIGUBES. NOS Plate X {opposite page 18.) Fungus developed in ordinary stool (muror). Appearance of the Mycelium on the second and third day ... xxviii. 1-2 A fertile filament which crept out of the preparation, and which bore a distinct cyst on the seventh day. Delined spores could not be distinguished among the contents ... ... ... „ 3 Growing-cell, in which is seen the position of the preparation through the thin covering-glass. Between this glass and the subjacent glass-slide the fungus (xxviii) above described was cultivated. The varying diameter of the segments of the circles enclosing the preparation permits the entrance of air ... ... xxix, Plate XI {opposite page 20.) Globules of a fatty nature simulating " cysts," " spores," &c. ... xxx. Greenish-yellow globules which formed a considerable portion of the sediment of a cholera stool ... ... ... xxxi. Spherical form of ditto ; the tinged portion is seeu to be contracted from the delicate pellicle which encloses it ... ?. xxxii, 1 Oval and irregular shape of ditto ?, ... ?. „ 2-3 Appearance presented by the foregoing in the course of four hours ... xxxiii. Vanished suddenly, a pale " ring" only remaining ... ... „ 1 Granular appearance which occasionally preceded this condition ... „ 2 Granular appearance of ring-like remains ... ... ... „ 3-5 Spherical body with a dense, tinged substance (oil) centrally situatctl „ G An aggregation of the foregoing globules surrounding a phosphatic crystal ... ... ... ••¦ ¦•• xxxiv. Plate XII (opposite page 22.) Microscopic appearance of a distended blood-cell at various distances from the object-glass ... ... ... ... xxxv. 1-5 Aspect presented by the blood-cell at the end of three hours ... „ G Blood-cells from a cholera stool ... ... ... ¦¦• xxxvi. Presenting a single hyaline protrusion, capable of being retracted ... „ 1 Presenting two retractile protrusions ... ... ... „ 2 The protruded portion after a time is frequently not retracted, but is seen to trail with the cell when the covering-glass is shifted, as long as the cell is visible ... ... ... „ 3 Blood-cells similar to the foregoing (xxxvi) observed in " Chylous" urine ... •¦• ¦•• ••• ••• xxxvii. Some of the aspects presented by these cells ... ... „ 1-5 Various forms assumed by one of the larger corpuscles present ... „ G Embryo of a Round-worm imbedded in a mass of gelatinised substance which formed in " Chylous" urine ... ... xxxviii. Embryo (of a larger sizo than that delineated at xxxviii). after the addition of acetic acid. The hook-like appearance is only evident in certain positions ... ... ... xxxix. 1 The caudal -Inirm which became evident after prolonged action of the acid ... ... ... ••• ••• » FIGURES. NOS. Plate XIII {opposite page 24) Tyaline appearance occasionally seen, when examined early, of the cells associated with the flocculi in rice-water stools ... xl. 'he granular aspect presented by the preparation delineated at xl after 24 hours ?. ... ... ... xli. .nimalculfß which appeared in the evacuation on the fifth day. These generally present a distinct nucleus and frequently two anterior filaments, which the animalcules figured in plates xv and xvi do not ... ... ?. ?, xlii. 'he flocculi and the cells imbedded therein observed to be granular, although examined almost immediately. The granular mass observed at the upper corner of the figure may be defined as a Micrococcus Colony, produced by the disintegration of the substance into molecules ... ... ... ... xliii. [ovements exhibited by the corpuscles associated with the flocculi when freed from the meshes of the membranaceous substance xli\ r , Appearance of the corpuscles associated with the flocculi after the addition of weak acetic acid and iodine .., ... xlv, Plate XIV (opposite page 26.) 'he elongated form very commonly observed of the corpuscles imbedded in the flocculi. Some are granular, others are hyaline ... ... ... ... ... x lvi. Appearance presented by the preparation (xlvi) after the addition of iodine solution ... ... ... ... xlvii. 'urcince, as commonly observed in cholera and other stools ... xlviii. (¦cumulations of a fatty nature ... ... ... xlix. 1 ittle pellets which possess the power of altering in form and position ... ... ... ... 2 forms assumed by one of the foregoing ?, ... ... „ Very active animalcule ... ... ... ?, , 4 arious forms assumed by the gelatinous-looking substance depicted at xlix, No. 3 ... ... ?, ] nimalculse in a globular " still " condition ?. ... li. 1 arious forms assumed by owe of the foregoing ... ?. „ 2-5 Plate XV {opposite page 28.) llustrating the various stages in the existence of the animalcule which have been observed in alvine dejections. he aspects usually presented by these animalcule when seen in evacuations ... ... ... iit ]jj c Appearance of the preparation delineated at figures xlix— Hi on the fourth day ; many of these jelly-like masses are animalcules which have become inactive ... ... ... liii. arious forms assumed by a single animalcule immediately before it became inactive, as at liii ... ... ... liv. fleet of re-agents on the masses depicted at liii ... ... Jv. fter the addition of acetic acid ... ... ... ?1-2 ' " ~~ absolute alcohol ... ... ... „ 3 ' " ether and alcohol ... ... ... „ 4 Berkeley's growing-cell ... ... ... ... ] v i. 13 DESCRIPTION OF ILLUSTRATIONS. 14 FIGURES. NOS, Three stages in the " life history " of the animalcule, above described, which were followed out by continuous observation in the Berkeley-cell ?. ... ... ... lvii. Plate XVI (opposite page 30.) Largo granular cells, amongst which very active animalculae are seen (cholera evacuation) ... ... ... ?. lviii. Cells associated with the foregoing (lviii) and closely resembling them, but exhibiting pseudopodial movements ... ... lix. With a single vesicle-like protrusion ... ... ... „ 1 Exhibiting protrusions from more than one portion of its substance. „ 2 The projected pseudopod appears to have passed through an external envelope in one case (3), whilst the projection seems to consist of the external layer itself in the other (4) ... ... „ 3-4 Projections which were no longer retractile ... ?, „ 5 A large corpuscle presenting movements of an amoeboid character ... „ G Blood-cells altered in appearance ; the result of osmosis ... „ 7 Animalcule (cholera stool) ... ... ... ... n 8 Blood-cells ... ... ... ... ... lx. l One of the blood-cells from the group (No. 1) altered in appearance by one of the animalcules ... ... ... „ 2-4 Animalcules with blood-cells intimately adherent to their substance. The animalculse in this case are somewhat larger than ordinarily met with (cholera stool) ... ... ... ... n 5 Appearance assumed by blood corpuscles from a healthy person, which had been added to a portion of filtered cholera stool ... Ixi. Stellate appearance of the red cells ... ... Mi „ 1 White corpuscles ... ... ... ... ?, „ 2 White corpuscles spread out like an amoeba ... ... ? 3 Subsequent aspect of the red cells. The condition usually observed when found in alvine discharges ... ... ?, „ 4 The alterations observed to take place in a single white corpuscle ... „ 5 White corpuscle surrounded by a halo-like pellicle ... ... „ 6 Corpuscles and animalcukc observed in the stool of a perfectly healthy person ... ... ... ... Ixi-lxii. As seen immediately after being voided ... ... ... ]xii. As they appeared 2 4 hours later ... ?, IM lxiii. Plate XVII (opposite page 38.) Illustrations of the development of the lowest forms of life. Monads ... ... ... ... ... lxiv. 1 Bacteria ... ... ... ... ?. „ 2 Vibriones ... ... ... ... ... n 3 Leptothrix ... ... ... . ... ?, n 4 Appearances presented in a filtered solution of organic matter (unboiled) on the second day ... ... ?, >># ] X v. Ditto third day, showing the appearance of the " heaps" ... lxvi. 15 FIGURES. NOS. Appearances presented in a filtered solution of organic matter (tinbuilcd) on the filth day, vibriones increased in length ... lxvii. Ditto ditto, circular bodies developed in the midst of the heaps lxviii. Developed on the third day in a solution of organic matter (hoiled) lxix. Plate XVIII (opposite page 42.) Illustrations of the development of the lower forms of life. Objects presented in a boiled and filtered solution of organic matter towards the end of the third week ... ... lxx. The animalculre present in the above solution, which towards the end of the fifth week could not be distinguished from those described as being present in the alvine discharges, both in the active and "still" condition. The nature of the green cells in the midst of the molecules is not known ... ... Ixxi. Spores developed in another test tube containing a portion of the organic solution used at lxix — lxxi. N. B. — This tube had been breathed into ... ... ... ... Ixxii. Ditto in process of germination ... ... ... Ixxiii. The appearance of the spores (Ixxii) as modified by the addition of gum water ... ... ... ... ... lxxiv. Dr. Maddox's slide for cultivation experiments. Two strips of tinfoil are seen to intervene between the glass-slide and tho thin covering-glass, with tho preparation in the centre. The arrows indicate the spaces left open for the admission of air ... Ixxv. Plate XIX (opposite page 44.) Illustrations of the development of low forms of life. Amceboid bodies which appeared in a boiled and filtered solution of organic matter on the fourth day ... ?, lxxvi. The various forms assumed by one amoeba ... ... ... \ A portion of the substance of the amoeba becomes detached ... „ 2 Tho detached portion exhibits movements ... ... q Process of division into two portions of nearly equal size ... „ 4 Segmentation complete ... ... ... M 1 r } Appearance of a contractile vacuolc in the detached segment ... q Ditto fifth day ... ... ?. ... lxxvii AmcL'bao 1 ••¦ » 1 Ditto becoming stellate on the addition of water ... ... ff 2 The form subsequently assumed by No. 2 ... ... ?, 3 The subsequent history of the amoebae (lxxvi — vii) ... ... lxxviii. Amujba) creeping across the field and discharging their contents ... „ 1 Ainteke, which became circular, and active movement was set up among tho aggregated molecules. A bright halo is seen to surround the globular mass ... ... ... pj 2 The halo disappears and tho contractile vesicle vanishes ?, „ 3 The mass becomes broken up altogether ... ... ... }) 4 Illustrating the changes which occurred in two solutions of organic matter obtained from the same source, placed on under two covering-glasses upon one slide ... ... ... lxxix-lxxxiii. Circular " yeast" cells and anguillulce ? which appeared in one of the preparations ... ... ... ... iit Ixxix. Developmental stages of a young paramecium ?, ?, lxxx, 16 DESCRIPTION OF ILLUSTRATIONS. FIGURES. NOS. Corpuscles developed in the midst of a heap of minute molecules ... lxxx. 1 Growth of the corpuscles ... ... ... ?, „ 2 A contractile vacuolo becomes evident ... ... ... „ 3 The animalcule after its escape from the corpuscle (3) ... „ 4 Irregular outline assumed by the animalcule in a thick fluid ... „ 5-6 The animalculso become encysted, and in this condition multiply by segmentation ; some are seen to exhibit contractile vacuoles, others not ... ... ... ... ... lxxxi. Plate XX {op>posite page 48.) A 33 r oung paramecium getting out of the encysted condition ... Ixxxii. Two encysted'^aramec&'a ; active movements were set up amongst molecules of the smaller one, and the cyst became detached from its fellow ... ... ... ... „ 1 The molecular contents is seen to have assumed the form of an animalcule, which, by its active movements, caused the capsule to become attenuated ... ... .., ... „ 2 The escaped animalcule ... ? . ? , ? , „ 3 The remains of the cyst ... ... ?. ... „ 4 A ruptured cyst — animalcule not escaped ... * ?. lxxxiii. 1 Animalcule escaping, but is still enveloped by a delicate capsule (schleier.) ... ... ... ... ?, „ 2 Empty cysts ... ?, ... ... ?, ? 3-4 Segmentation into four animalcules has occurred in the cyst ... „ 5 After several encysting processes, a ciliated infusorium appeared on the slide ... ... ... ?. ... „ 0 Forms of life which developed in a cholera stool ... ... Ixxxiv.-ix. The animalcule described as occurring in alvine discharges in the active and in the " still" condition ... ... ... lxxxi v. ElFect of the addition of carmine solution upon the above prepara- ™ tion, everything in the field being tinted pretty much to the same extent ... ... ... ... ... lxxxv. " Yeast" cells which appeared in the midst of the foregoing on the third day ... ... ... ... ... lxxxvi. Fertile filaments bearing Sporangia with spores ; the latter were readily distinguishable about the fourteenth day ... ... lxxxyii. Plate XXI (opposite page 52.) Fungi developed in cholera discharge. Earlier condition of lxxxvii : the filaments are intersected by those of Penicillium ... ... ... ... lxxxviii. Penicillium viride ?. ... ... ... ... „ 1 Penicillium glaucum ... ... ... ... „ 2 A more fully developed specimen of lxxxvii. Some of the filaments are seen to present dilatations or macroconidia ... ... lxxxix. Plate XXII (opposite page 66.) Objects observed in some moistened soil from Allahabad on the third and succeeding days ... ... ... ... xc.-xci. Various stages of Mbnas lens principally ; observed in soil at a depth of four feet from the immediate vicinity of the newly erected barracks ?, ?, ?, ?, xc. 1-8 DESCRIPTION OF ILLUSTRATIONS. 17 Fioubes. Nos. Minute Zoospores, together with animalculae in the " still" and active condition, precisely similar to those described as being present in alvine discharges. Developed in moistened soil obtained from the ilooring of the Clydesdale Barracks at a depth of four feet.. xci. Developed in moistened soil from Lucknow ... ... xcii.-iii. Tanophnjs in two positions ... ... ... ... „ 1-2 Euglcncß or Astasice ... ... ... ... „ 3-4-6 Ampliileptus ... ... ... ... ... „ 5 Two Moners are shown in the act of creeping across the field. One is seen to curve its Pseudopoda around the circular cells present — the encysted condition of some animalcule. A ciliated infusorium may also be observed in the figure ... ... xciii. Developed in moistened soil from Fyzabatl ... ... xciv. Zoosporoids ... ... ... ... ... ? 1-2 Monas lens ... ... ... ... ... „ 3 Paramecium (?) ... ... ... ... ... „ 4 Coleps hirtus ... ... ... ... ... „ 5 Developed in moistened soil from Meerut ... ... ... xcv..xcvi. Alyce ... ... ... ... ... ... xcv. 1-2 Monas lens undergoing segmentation ... ... ... „ 3-7 Plate XXIII (opposite page 74.) Various stages of Monas lens ... ... ... ... xcvi. 1-6 Euglenw(?J ... ... ... ... ... „ 7-8 Developed in moistened soil from Peshawur ... ... xcvii.-cvii. Spore of Helminthosporiuin (?) ... ... ... xcvii. 1 Monas lens ... ... ... ... ... „ 2-3 Various forms assumed by one amoeba ... ... ... „ 4 Panophrys in various positions ... ... ... xcviii. 1-5 Amphileptus ... ... ... ... ... „ 6 A Paramecium dividing ... ... ... ... xcix. 1-4 One of the segments after complete division : the arrow indicates the direction of the current ... ... ... „ 5 Minute Monera presenting no nucleus nor contractile vesicle ... c. A Moner throwing out Pseudopoda in all directions. A great number of vibriones are seen in the field ... ... ci. Plate XXIV (opposite page 76.) Various forms assumed by a single Moner in the course of two minutes. The vacuolye are not permanent, nor do they appear rhythmically. The coloured granules are drawn into its substance during the retraction of the pseudopods. The engulphed granules flow in the direction of the projected part, as indicated by the arrow at cii. and cv. ... ... ... cii.-vi. Two Monet's which have become spherical and still (under a lower magnifying power) ... ... ... ... cvii. Maps. Allahabad, opposite page ... ... ? , ... 62 Lucknow, opposite page ... ... ... ... 68 Morar and Gwalior, opposite page ... ... ... 72 and the magnifying power used is attached to each figure. yie diameter of the object in any of the figures may readily be obtained by ing them with the one-thousandth of an inch scale placed at the foot of each These plates have been engraved in the Office of the Surveyor General of India. They are very faithful copies of the original drawings, and will bear favorable comparison with the work of engravers in Europe, who are habitually engaged in this kind of employment. lam under great obligation to the Surveyor General and also to the Assistant Surveyor General, Captain W. G. Murray, under whose immediate superintendence the work has been done. INVESTIGATION INTO THE MODE OF ORIGIN AND SPREAD OF CHOLERA. PART I. CONCERNING THE THEORY OP THE FUNGOID ORTGIN OF CHOLERA AND THE MICROSCOPIC OBJECTS FOUND IN CHOLERAIC EVACUATIONS. The theory of the fungoid origin of cholera is based upon result of certain experiments instituted by Professor Hallier, with the view of ascertaining whether any special organisms could, by means of artificial cultivations, be obtained from choleraic discharges. These experiments have been repeated many times in Calcutta, but as the daily notes [of each cultivation would occupy so much space, I propose giving a short summary of a few of the cultivations, illustrated by some of the camera lucida drawings which have been accumulated during the investigation. It may, however, previous to doing so, be well to state, in as few words as possible, Avhat the theory really is. As the Professor has published the result of his labours, a short epitome of his brochure, weeded of as many technicalities as such a subject will permit ; together with a selection of the leading figures in the plate, attached to the book, will, it is thought, best serve to convey his meaning. Some choleraic discharges ivere sent to the Professor ¦ o at Jena, obtained from a patient at Epitome of Hauler's cuiti- Berlin during the epidemic of 1866, tTeatw CC ° mpiled tr ° m Ms and another specimen from a patient at Elberfeld during the epidemic in * " Das Cholera Coutayiuw." You Dr. Ernst Hallier. Leipzig, 1867. 2 A SUMMARY OF HALLIETt S VIEWS 1807. These were examined microscopically and found to contain : — 1. Cysts of a yellow or brownish colour, which he for some time believed to be the fruc- Discovery of cysts in cho- tificatioil of UVOCyst'lS / SOUIC of these lera discharges, which seem- j | c irregular OutlillO, UJld at fid at first sight to present . «' S? no organised structure. first Sight Seemed to pOSSCSS 110 Organic structure, caution being necessary not to confound them with masses of fat ; application of pressure was, however, found sufficient to discriminate between them. A drawing is given of some of these in a swelled, broken up condition (Fig. i, 1). 2. Here and there a few other cysts were seen, con- sidered to be of the same kind as the Other cysts more distinctly forgoing . feW WCI'C spherical 01' OVal organised. o © ' i 7 t X cysts varying considerably in size, enclosing a number of yellowish shining spores ; the spores also varying in size, as may be seen by a reference to the accompanying figures (Pig. i, 2). 3. Groups of swollen gelatinous spores surrounded finely molecular matter (Fig. i, 3). spores of foregoing in a others 'appear granular, and some swelled condition, y l ° n . show indication of fission. 4. MicrococoMS. — The molecular matter just alluded to, supposed to have originated from "^rooootm." down **** the breaking up of the plasma in the " spore," a little heap often being observed, corresponding to the previously existing spore, called a " micrococcus colony" ( Pig. i, 4 a), which at b is still further broken up; at c a group of "colonies" is seen corresponding to the mass of spores previously contained in a cyst whose walls have disappeared. The minute protoplasmic molecules constituting these colonies were seen to adhere to various objects in the fluid, and espe- «££Sta£ micrococcus on dally to the particles of epithelium, in fact feeding upon them ; this being invariably the way in which vegetable parasites first attack animal tissues. In the midst of these molecules larger ones were observed (Fig. i, 5), which have of micrococ- j^ figured mft stiU mQpe advanced stage as torula-like formations at 6. This condition being, according to Professor Hallier, the transition stage to the development of the higher forms of fungi. A series of cultivations was carried out in order to prove 3 S TO Tllljl* Xi A^XoTllj^l \j Vj \Jj* -A. C IJ-OJjlii XV .A. J\j N (jr LJ o« that these bodies were organically related to each other, namely, that the irregularly defined cultivations instituted to cysts were advanced stages of the cyst rr e ;rtnfSr gg O oTnT tiOnbe " with sharp contour and well marked spore contents; that the circular gelatinous-looking bodies were originally contained in capsules; that the capsules had been borne on a filament; that the filament had originated in a " micrococcus" cell ; and that the " micrococcus" had been derived from the disintegration of these gelatinous spores. Portions of the discharges in question were isolated, and placed upon various substrata, beet*, starch -paste, slices of lemon, &c, so as to supply the "micrococcus" with other nourishment than the epithelium of the intestinal canal, the disorganization of which substance is, according to Professor Ilallier, the prime cause of cholera. The results of these cultivations may be thus briefly described. During the first two or three Besult of the cultivations. i jt • • n i days the micrococcus rapidly increased in amount, and developed into nucleated cells, which arranged themselves into chains, as already observed to exist to a slighter extent in the original evacuation. In some cases a thin pellicle formed {mycoderma), which, on being lifted, fre- quently broke down into round balls Germination of micrococ like the ' IIIicrOCOCCUS Colonies ;' the 0 11 M. torula cells about the fourth day were seen to germinate, the ends of the filaments having a linked appearance (6), which continuing to grow, presented the appearance usually seen in oidium lactis. In the course of a week the filaments assumed a Development of filaments branched and saeculatcd appearance with formation of macroco- /t:v • /-? r\\ j_i i • • j nidia . (-tig- !> 7, 9), these saccules or joints (termed " macroconidia") being capa- ble of germinating like the spores. The spores were on several occasions seen to produce a peculiar form of fructification, considered by Dr. Ilallier to be degenerated Tillelia caries (smut), (Pig. i, 9), and on one occasion a spore somewhat like that of smut was detected (sp.) ; a few S^SSSiJSSS 1 abOTtive attempts at the formation of spore-containing-cysts were also seen. In a few instances, however (about the 9th day), the filaments were observed to bear unmistakable .cysts, some with the contained spores very evident (Fig. i, 8), and others in which this condition was less clear. 4 SECTION I. — " CYSTS." The nearest approach to the development of the cysts, corresponding to those in the The extent of cyst deve- di sc h ar cr es wliich reminded the Prolopment. . . __ __ ___ _ ? . fessor so strongly of Urocystis, is figured (Fig. i, 10), and the germination of the same at Eig. i, 11. The inferences drawn by Dr. Hallier from these experiments in a few words are, that cholera is produced by a species of fungus belonging to the us- Importance attached by UlaghieS Or SHlut GTOlip. TMs flingUS Professor Hallier to these • i j. • •¦• 1 rt ± j.x__l cultivations. 1S a polycystis, similar to that attack- ing the rye only in Europe, but which the Professor believes attacks the rice plant in India ; grounding this belief on the fact that, in the tissue of growing-rice plants watered with choleraic discharges, bodies were detected which he con- ing^wSS pTan t r lat " sidered identical with the cysts found in the evacuations, thus accounting for the belief frequently expressed by the older writers, that cholera was generated by the consumption of rice in a diseased condition. The author has since modified his views as to the species of fungus in question, but retains the opinion that, whatever the fungus may be called, it closely corresponds with the fungus observed to develop in soil contaminated with choleraic discharges. It will now be seen that Professor Hallier believes that he has established an organic connection between the two kinds of " cysts," " spores" and " micrococcus." The questions naturally arise — (1) Are there such bodies in the choleraic discharge examined in India ? (2) What are they ? and (3), are they found under similar circumstances elsewhere ? SECTION I.—" CYSTS." Dr. Hallier appears to have derived the first idea of cholera cysts from the engravings of the " cholera bodies" of Drs. Swayne, Erittan, and Budd, in the year 1849, as reproduced in M. Hobin's work on Vegetable Parasites.* For, after stating that they are un- Tiie •• cholera cysts" flgnr- doubtedly of the same nature, indeed in M. Robin's work on • n n -. . *> ,-i "Vegetable Parasites" iden- m S "'Om the drawings, of those tilled by Haiiier. seen by him. a severe renroof is seen by him, a severe reproof is administered to the Erenck author * Hlstoire Naturelle dcs Vegetaux Parasites. Atlas, PI. XII., Figs. 4-5. 5 CHOLERA BODIES OF DRS. BUDD, BRITTAN, AND SWAYNE. for the summary way in which he disposed of the " cysts" of the Bristol Doctors. As these " cysts" have been the subject of discussion for more than twenty years, without any definite conclusion as to their real nature having been attained, a few observations History of these bodies. coucemhlg them may not fee Ullinteresting. In September 1849, Dr. Brittan published a description of the bodies observed by him, termed " annular bodies," in the London Medical Gazette; this term comprisin 1 bodies varying considerably in size and appearanc — large masses corresponding to Hallier's cyst, and smaller bodies which probably correspond to Hallier's spores. Mr. Brittan did not attempt any cultivation so as to connect the one class with the other, but inferred that they were the same in different stages of development, because he had observed something like a connection between the size with the severity and duration of the disease. The late Professor Quekett, of the Royal College of Surgeons, coincided with him in the belief that they were different stages of the same body, and of a fungoid nature. Mr. Swayne also announced that he had discovered certain cyst-like bodies which were named " cholera-cells, " drawings and descriptions of which he published in the Lancet about the same time as Mr. Brittan. He also believed that the larger and smaller bodies figured were mere stages in the development of the same thing. Dr. Budd believed that he found similar bodies in the water of tainted districts, and designated them " cholera fungi." These announcements caused considerable excitement at the time, which was somewhat lessened when .^SS^SEJ&iS! 0 Mr. Busk announced that the bodies in the sample received by him were a species of uredo ( TJredo segetum), the bunt of wheat, illustrating his statement by the removal of bodies like the one in question from a loaf of ordinary brown bread. The College of Physicians appointed a Committee of Inquiry, and Drs. Baly and Gull drew up a report, in which the small bodies are said to be either carbonate of lime (probably from the aromatic of Drs. Baly and Gull. confection mixture taken), disintegrated blood-cells, or starch particles ; the larger ones figured by Dr. Budd to be probably accumulations of starch cells with disintegrated particles of vegetable tissue, and those of Drs. Brittan and Swayne to be some species of bunt, as identified by Mr. Busk. The 6 THE VARIOUS CYST-LIKE BODIES Reverend M. J. Berkeley, the greatest authority on fungi we have, on being referred to, declared bSLi^. Eeverend M< J< that the specimens he received were not fungi at all, so that evidently the propounders experienced some difficulty in recognizing their own " bodies," otherwise such microscopical experts as Mr. Berkeley and Mr. Busk would not have been supplied with such cntirelv different substances. Here the matter rested until Professor Ilallicr observed a resemblance between the cysts in Hn°-n,H n°- n , i or reproducine the choleraic discharges examined by M. Eobm's figures. © . . y him, and those figures in M. Robin's book, which figures are here reproduced (Fig. ii), as being the only criterion we possess of what ilallier really means when he speaks of cysts ; the only drawing published by him of the mature cyst being that of a ruptured one (i, 1). In the examination of cholera dejecta which I have made in Calcutta and in the North - Cyst-like bodies in the Western Provinces, mailV " CYSts" elioleraic dejecta observed -> -i -i ,-1 " in India. were observed, and these in many cases closely resembled the ones eJ figured in M. Robin's work, but were not of such universa occurrence as the attention they have obtained woulc have led one to expect; indeed, frequently absent altogether. The "cysts" figured by Drs. Brittan and Swayne (the greater part of which are here reproduced from the drawings accompanying the original articles of these gentlemen) arc certainly the kinds most frequently present in evacuations, as the fact that the following observations concerning them were completed before either the original figures or copies of them had been seen, would tend to show. They may be divided into two classes. The principal figures in Dr. Brittan's drawing will two distinctly different se rve as an illustration of one kind kinds of "cysts" figured by /n^. ... _ N , , , ? „ Brittan and Swayne. (ilg*. HI, 1), and tllC leading figures m Dr. Swayne's of the other (Pig. iv, 1-4). As the two classes arc copied in M. Robin's work, and Dr. Ilallier does not intimate his belief that they are not of the same nature, it will perhaps be best to allude to the two, so as to leave no stone unturned in the matter. That they vary much in their nature will be manifest from the following observations :—: — 1. The dejecta of a patient who had been suffering OBSERVED IN CHOLERAIC DISCHARGES. 7 from cholera about 12 hours, and who died on the second day, presented an enormous quantity Irregular dark-yellow () [ o>l o blllar masses of a dark-yellow "cysts" corresponding to .° , , " , Brittan's "annular bodies." COIoUT, CXCept at tllC CCntrC, where the colour was much lighter, and the mass was much more transparent than at the side (Fig. v, 1), which is not unlike the ones figured Action of liquor potass*; fey gtpong Hquor potass£B being added, one of the " cysts " burst, as at 2, and gradually broke up, as at 3, 4. The semi-fibro-gelatinous mass in which the " cysts" were involved was entirely dissolved. Another slide was taken, and two cysts selected, a large one and a smaller one (Pig. vi, 1, 2) ; strong acetic acid being added, no result followed for some time ; of acetic acid; pressure was applied, and the large one broke up into air-globules and granular matter (3, 4). Pressure was again applied, and the small one also broke up, as shown at 5. Another case may be quoted as illustrative of this kind of cyst. The evacuation was passed six hours after attack, and two hours before death ; it contained numerous cyst-like bodies, some entire, others more or less broken up, and in many cases seemed to contain partitions (Pig. vii). These bodies of ether. withstood the action of rectified ether until the fibro-albuminous matter surrounding them had been, removed by the application of potash. 2. In the same evacuation other globular cyst-like bodies were found of a yellowish-green tint, wen defined, round or ovai having a more defined outline, and greenish-yellow cysts un- mQre even ly diffused COlltcilts (Pig. affected by ether and liquor ... . " ml „ \ .° potasasß, resembling Swayne's VJII, 1, 4). TIICSC Were Ulianected by cholera-cells. +1^ oflim> nnrl vnrnninrvl lmnffpofnrl lw the ether, and remained unaffected by liquor potassse for three days. These cysts occurred in nearly all the evacuations examined, but their precise nature was for a considerable time unexplained. They were sometimes round, but generally oval, and in some cases formed about a fourth of the entire sediment. This was particularly observed in some dejecta with which I was favoured from the Medical College Hospital, obtained from a native who was admitted with all the symptoms of cholera, but eventually recovered. They were, as in other cases, of a greenish-yellow* tinge, with colourless hyaline capsules, for the most part oval (x) — sometimes round, and varying considerably in size, as seen in the figure. One of these cysts was 8 THE VARIOUS CYST-LIKE BODIES selected for special observation*, the one represented at Pig 1 , ix, 1 ; ether being added, the contents cleared up a little, bui nothing further ; this was followed by strong liquor potassae, which caused it to become dotted and streaky, the yellow tinge, however, remaining (2 ) ; gra- pettedT ° f re ' agents re " dually changing to the appearance depicted at 3, the centre becoming more transparent than the circumference, which still furthei extended, as at 4. The transparency of the central portioi diminished in the course of a few minutes (5), in which con dition the object was left under the microscope until the nexi morning, when it was found to have retained its form, but hac acquired a dark colour. Another cyst was selected with a dark-yellow granular centre, and hyaline capsule (Fig. x, 1) Ether was added; scarcely any change; Sv?d° material change ob " merely clearing up the centre a little. It was then rolled over, and the granular contents spread throughout the entire cell (2). Firm pressure was applied, the eye being kept steadily at the microscope, when suddenly numerous minute molecules es- caped (3), and the capsule became partly emptied of its contents (4). Resnlt of pressure. Liquor potassec was added to a portion of the evacuation and allowed to stand all night. The cysts on examination next morning appeared unaffected. To another test tube sulphuric acid was added. The cysts after remaining several hours in the acid were not much altered, but presented a globular outline with a hyaline capsule surrounding a greenish-yellow molecular mass (Eig. xi, 1, 2). On rolling them over io^T^i^™ they became oval, but soon regained the circular form. On the addition of a strong solution of iodine, the contents became dark-brown, and on subsequently adding absolute alcohol, fat-like globules made their appearance, which, by manipulation, could be made to move within the cell ; the capsule being unaffected (Eig. xi, 3). Alcohol being added to another cyst without the iodine, the contents assumed a lumpy appearance with a clear space in the centre (Eig. xi, 4) . Several very small embryos of round-worms having been observed in the evacuation in active motion (Eig. viii, 8), diligent search was made as to their ciat^ththec^tT 8 aSS °' ™g™> which resulted in the explana- tion of the nature of the cysts also. OBSERVED IN CHOLERAIC DISCHARGES. 9 The latter were frequently observed to give evidence of some kind of systematic arrangement of their contents, as shown in the figure (viii, 5, 6), and eventually a cyst was observed to contain something which rolled within it ; this, after prolonged watching, Contents of the cysts. was seen to present the exact form and size of the worm-like body just alluded to. It was coiled up on itself within the capsule (Eig. viii, 7), and continually altered its position. This corresponds pretty accurately with the drawing of the ovum of dscaris my s tax in Dr. Cobbold's work on Entozoa. It is, I think, pretty much the same as the cholera-cell of Mr. Swayne. In many cases the contents of these ova are also shrunken, occupying a part only of the enclosing membrane, as insisted on by this gentleman as a means of diagnosis. The effects of re-agents also, as above given, correspond very closely with the description given by him. 3. There is another cyst not very uncommon in choleraic dejecta, having a more delicate, but very resistent capsule (Eig. xii). Its nature may be inferred from the following statement : On two or three occasions, Acan, and their ova in scm i_di sm tegrated acari were obcholera stools. , . ? o , . , ?, served in the stools examined, which had, in all probability, been swallowed with the food, in bread perhaps, and passed through the intestinal canal without being very much broken up, as may be seen from the figure (xiii). It did not, however, occur to me to connect the existence of the thin capsuled cysts with these acari, until one day two were seen rapidly depositing their eggs among some fungi under cultivation, which were being microscopically examined. These eggs corresponded precisely with the just described cysts. 4. Mr. Brittan figures some oblong bodies (Fig. iii, 2), which are not reproduced in M. Robin's plates, but were probably also considered to have some connection with cholera by the author of the article in the Medical Gazette. These are exceedingly common, and are accurately drawn in Pig. xiv, where one is seen entire, and another ruptured, to- gether with one of Mr. Swayne's bodies mOßrittiXdmwTm O BrittiXdmwTn U g nd ' WOrm in a m P turec [ condition ; both required the application of considerable pres- sure before the capsule gave way. The first described elongated body is, I believe, the ovum of another round-worm, the Tricocephaliis (dispar?). As to the cysts with distinct spore contents, which Hallier has figured (Eig. i) as being a mature v 10 THE VARIOUS CYST-LIKE BODIES condition of the cysts comparable to the drawings in Robin's work, I have not met with any which were unmistakably the same in fresh dejecta, but have developed them repeatedly ; the particulars will be given further on. Other cyst-like bodies are occasionally found, but as they do not in any way correspond to those of the author of the theory under consideration, a description of them is reserved for another occasion ; the principal ones, however, are those summary of the principal already described, namely, (1) comcyst-like bodies observed in i j •*v dn. ± a cholera. pound cysts, consisting oi fragments of various tissues and fat surrounded by a semi-organized fibro-albuminous layer, and (2) ova of various kinds, none of which are peculiar to cholera. As, however, the ultimate elements of other cysts than these might exist in the dejecta, every known method was resorted to for the purpose of developing them, a few illustrations of which I give in a condensed form. Illustration I : — Small portions of the dejecta which contained such numbers of the cysts, alluded to in page 7 and represented at Fig. v, were placed in three perfectly clean watch-glasses with the following substances :—: — I. — Cholera evacuation 3 drachms, and 2 drops of acetic acid, so as to neutralize it. II. — Cholera evacuation 3 drachms, phosphate of ammonia 3 grains, grape-sugar 3 grains. 111. — Distilled water 3 drachms, phosphate of ammonia 3 grains, grape-sugar 3 grains. To receive these, a small wire stand had been placed in a shallow dish containing a strong solution of permanganate of potash, and the stand and watch-glasses covered in by a bell-glass (carefully cleaned, and subsequently rinsed with alcohol) which stood in the fluid. This was set aside in an average temperature of 82° Fahr. On the third day small white specks were seen on the surface of No. I, which had returned to its alkaline condition, one of which was picked out as rapid- Progress of the cultivation. , b j Ma * f r n ru +T _ /ii ly as possible from beneath the bell- glass and placed on the stage of the microscope. It consisted of an aggregation of minute molecules held together by a slimy substance, from which filaments of fungi escaped (Fig. xv). Thus matters stood until the fifth day, when from No. II being picked out, presented numerous spores 11 OBSERVED IN CHOLERAIC DISCHARGES: CULTIVATION. (Kg. xvii, 1), many of them germinating very actively (2), and the filaments here and there were swollen out into macroconidia (3, m) 9 some of these dilatations being transparent, others ffranular ; frequently the filaments were seen to terminate in a bulb (4), and in one case a filament was tipped by a cyst in which the contents One cyst developed. were granular and had contracted from the capsules (5). Pre„ - i i i>i i j • _ 1} 1 ' TVT^v cisely similar filaments and dilatations were found in No. I, but a distinct cyst (or sporangium) could not be seen. This condition lasted until the seventh day, After the seventh day asper- wnen the mycelium gradually degegillus illus appeared in all. & appeared on all three, of various colours, but principally of the dark varieties. Illustration II: — ¦ A portion of the fluid contents of the small intestine from a patient who had died within six hours of attack was ¦L /* -i -i i nil •_ 1 J carefully transferred to a vial, and allowed to settle for an hour. In the Growing solution. meantime a " growing" solution was made, consisting of grapesugar 3 grains, phosphate of ammonia 10 grains, glycerine 1 drachm, and distilled water 1 ounce. A drop of this was placed on three glass slides ; to these were added :—: — No. I. — A minute quantity of the upper layer of intestinal contents. No. II. — A minute quantity of the sediment chiefly. No. 111. — A minute quantity of diabetic urine containing " yeast cells." These were placed as before under a bell-glass placed in Condy's fluid ; on the third day specks appeared on the prepara- tion in each slide, which proved Bpores germinating in each t ft Q fa spores an d mycelium preparation on the third day. . . , f ?, .. (Fig. xvi), the three slides presenting similar appearances under the microscope. On the fourth day No. I presented an excellent forest of penicillium, and No. II a similar crop of aspergillus, of the B e°Su!™ e " ™ d '" W a(!k and PMjto coloured variety, while No. 11l produced both penicil- lium and aspergillus. These were systematically examined for eight days, no other fungus making its appearance. The aspergillus crop in No. II presented tufts of different colours ; specks were observed in the other two preparations ; a speck of yellow and brownish-purple being the most abundant. 12 CULTIVATION OF CHOLERAIC DISCHARGES To experiments conducted in this manner, there is the serious objection that each time the preparation is examined, no matter how carefully, the possibility exists of foreign matter getting into the preparation. With the intention of obviating this source of fallacy as cribed" tinS apparatus dcs " much as possible, an aspirator was em- ployed to supply the preparation with purified air, at least as pure as passing it through concentrated sulphuric acid Avill allow. By referring to the accompanying sketch, it will be readily seen how this was effected (Fig. xviii). A small funnel (1) with a pledget of clean cotton wool inserted into its neck was attached to a piece of bent glass-tubing ; this tubing passed through a perforation in the cork of a flask (2) containing concentrated sulphuric acid ; from the neck of this flask another piece of glass-tubing emerged which connected it with a perforated bell-glass, standing in a shallow dish containing Condy's fluid ; (3) another piece of tubing connected this with the aspirator (4) filled with water. All the connections were carefully luted, so that the only air which could have got at the preparation on the stand within the bell-glass (of course minus the air which previously existed therein) must have passed through the sulphuric acid. Illustration 111: — A perfectly fresh choleraic evacuation having been obtained two hours before death (in a rapidly fatal case lasting only seven hours), three watch-glasses were placed in the isolating apparatus with the following ingredients :—: — No. I. — A slice of the interior of a plantain weighing quarter of an ounce was scooped out, J£S££Z££? " thC and six dr °P s of tlie sediment from the evacuation was placed in the little cavity thus made. No. II. — A few drops of the evacuation-sediment only. No. 111. — A slice of the same plantain as in No. I. The apparatus had been made as clean as possible previous to this, rinsed out with spirit immediately before depositing these glasses on the stand beneath the bell-glass, and the greatest care taken to avoid foreign matter getting at the preparations before placing them there. The air within was renewed morning and evening ; the weather was warm the whole time, the average day temperature of the room being about 90° Fah\ FIG XX MYCELIUM WITH MACROCONIDIA BUT WITHOUT SPORANGIA \ On the fourth day a mould was seen to appear on the two slices of fruit, quite as marked allowed to remam m the Q foe clean plantain as on the other, apparatus for three weeks. * , . * but no change was visible in the watch-glass containing the evacuation only. This condition lasted a fortnight, the crop of fungus gradually increasing in the two former, and no change could be observed in the latter. During the third week the fungus not having made any progress, and the liquid in the watch-glass No. II becoming rather less, from evaporation, the apparatus was opened on the twenty-fourth day, and the result carefully examined forthwith. The two pieces of fruit were covered with a thick coating of a black and yellow coloured fungus, re^vai. ion ° f the *"" ° n both colours appearing in the two preparations; the yellow prevailing in the tainted slice, and the black on the other ; the difference being merely in the proportion, for tufts of each colour appeared here and there over the surface. These were found under the microscope to be aspergillus (Pig. xix, 1) and penicillium (xix, 2). Precisely the same fungus and the same species grew on glycerine, on starch-paste, and on pieces of dirty cork in various parts of the room. In the other watchglass, however, containing the evacuation only, a very different appearance was observed. The preparation had become partly dry, and presented a filmy appearance. On placing the watch-glass on the stage of the microscope, a great quantity of spherical bodies were seen The fungus developed in w ith granular contents, the average the watch-glass with evacua- • -i • -i . ,t , n 1 >j_ I•% i lion only. size being about that oi a white blood- corpuscle, but the size varied consider- ably, among which long delicate mycelical filaments ramified (Fig. xix, 3); from this network thin fertile threads arose, tipped in most instances with exceedingly delicate vesicles (xix, 4), which appearance at first was taken for the dew-drop aspect so common to mycelium ; others were seen of a much larger size. On watching them closely, all the bodies were seen to roll round and round, like a volvox. Elongated (spore-like) bodies were distinctly visible within each delicate _ capsule, unless very small (xix, 5), Description of the delicate j t , " • ±- n cysts. and seemed to move irrespective of the capsule (or sporangium) : of this, however, lam not certain. They appeared white by reflected light, and yellowish- green by transmitted light. The 13 14 NATURE OJ? THE rUNGI DEVELOPED. movement appeared to me to be due to currents of air in the joom, each little sphere twirling round rapidly in one direction for ten or twenty turns, then as rapidly twirling in the opposite way. The course of the spinning vesicle was not always horizontal, but varied until it was nearly vertical to the filament on which it was perched, but never quite vertical. It seemed analogous to the spinning of a plate or ball nicely pivoted on a juggler's stick, which may be seen to revolve in every direction but the vertical, the analogy being complete, except that the organic connection between the sporangium and the stalk rendered reverse turns necessary. On touching this with water, the capsule appeared to become instantaneously dissolved, no trace being left : the spores had fallen down, and the filament looked perfectly bare. Some parts of the mycelium were dilated into saccules (or macroconidia) (Fig. xix, 6), but no evidence of spore contents Avas distinguishable. Illustration IV: Being desirous of ascertaining whether from the rice- water stools in epidemic cholera I cultivation of a cholera C0ll \ ( \ produce capsules more unmisstool obtained during an epi- i i ti ,i ,» -it -n n demic or the disease. takably like those figured by rroiessor Hallier than I had succeeded in doing from discharges obtained in an endemic locality, such as Calcutta is, a sample was brought from Lucknow, carefully secured in a clean vial, which was obtained during my visit to the North- Western Provinces during the epidemic of cholera which occurred there in September last. A drachm of the sediment was poured into a perfectly clean watch-glass, and placed on the stage in the isolating apparatus in the manner described in the last illustration. In the course of a week a film was seen to have formed, which continued to increase in density for another week, but no trace of any mould could be observed in it through the bell-glass. At the end of three weeks the preparation was taken out and microscopically examined, but no cysts had formed, as in the former preparation treated in exactly the same way, but there was a great quantity of mycelium, in the No cysts were developed, rnpshpfi nf wTu'pTi TininorniKS oirnnlar but a quantity oi' segmented mL » 111^ Oi WUIOII numCIOUS CllOUldl mycelium. bodies were embedded (Fig. xx) ; the latter seemed to be the result of segmentation of the former, judging from the similarity FIGS: XXI XXIII. DEVELOPED IN ORDINARY EXCRETA between the free cells and the imperfectly detached segments of mycelium. The watch-glass was replaced in the apparatus for a fortnight, but no change took place. Prom these illustrations it will be seen that whereas cysts, distinctly resembling those described by Professor Hallier, may, by cultivation, be observed to develop in choleraic discharges, yet they are by no means constantly obtainable, for out of more than a hundred cultivations, made with the express object of developing these cysts, only three times was I able to produce any fungi bearing such tokens of fructification. Is it possible to develop fungi in other than cholera dejections bearing fruit resembling the " cholera cyst ?" The answer must be "Yes," as the following experiment will show :—: — Illustration V : — About half an ounce of fteces, obtained from a perfectly healthy person, was placed on a Cultivations of ordinary sma H on] ass plate, and carefully transhealthy stool, one isolated „ i • j ' n in -i n n and the other exposed. ieiTed into the DCll-gIaSS 01 tlie isolating apparatus in connection with the aspirator, as already described, the greatest possible care having been taken to prevent foreign matter coming into contact with it before dejiositing it on the stage in the apparatus. A small portion of the same substance was placed on a glass slide, without any special precautionary measures being taken to prevent access of foreign matter, so as to be able to examine it from day to day for comparison with the preparation in the bell-glass, which it was not intended to disturb. On the second day a few small white spots were observed on both preparations, one of which was picked out with a needle from the Jroaressofexposedprepar- non . isolated ma§S} and placed Qn fa e stage of the microscope. It consisted entirely of minute molecules, round and elongated (Pig. xxi, 1), embedded in a white shining substance (2), in connection with which were circular and oval cells of a greenish tint (3) ; frequently two or more were seen strung together (4) ; clear spaces were seen in them all nearly. On the fourth day the mass in the apparatus was completely coated by this white humus, except that some of the earlier observed spots had acquired a yellowish-brown colour. The exposed slide presented a somewhat similar 15 16 THE CHOLERA FUNGUS OF TnOME* AND OF HALLIER appearance. The cells had become nearly everywhere strung together, and long filaments of oidium Th?m.." cholera funsus " ° f lacfis (p( pi n- xxiii), corresponding ex- actly to the figures given by Thome of the cholera fungus discovered by him, to which rather a long name was given at the time, viz., " Cylindroicenium C holer cc Asiatics." This condition lasted till the sixth day, when a crop of a white mould was perceptible in the isolated preparation, and a plentiful crop of penlcillium and aspergillua appeared on the other cultivation (Fig. xxii). This slide having become rather dry, a few drops of distilled water were after this occasionally added. On the eighth day long delicate li la- ments were seen growing out of the Condition of the prepara- white lmmUS-lookillg SUbstaHCC ill tion under bell-glass after ,-, , in , i ¦, first week. the apparatus, and on the tenth day other filaments were observed, which seemed to be tipped with various coloured heads, apparently of the same kind as on the other slide, those of a bluish and yellowish -brown tint prevailing; but by the eighteenth day the long delicate filaments had grown over them, the whole surface of the preparation presenting a woolly appearance. After this no further change could be seen to take place in either cultivation, The apparatus opened on i fi j. .j. i> i. i ej i the twenty-first day. ana on me wenty-HISI (Liy <>l UK experiment the bell-glass was opened, and the glass plate placed on the stage of the microscope. Precisely the same species of aspergillus and penicillium were found as existed in the non-isolated cultivations, with the addition that great numbers of the filaments forming the white flocculent tuft bore at their terminations cysts or sporangia filled with distinct spores (Pig. xxiv, 1-4), which, I think, correspond exactly to the Cy »\ ob Itained1 tained ?» ct i y lik ! cysts figured by Professor Hallier the "cholera-cysts" figured J ? , O • , , , by Haiiier. of the immature cholera-cysts, whose drawing has already been given and may be compared with this.* Aspergillus tufts were present in great numbers : nearly all of them had fallen off from a germinating aspergillus filaments among the mycelium ; tuft simulating a " cholera ~ n 1 cyst" in the same condition: a few, IIOWCVCr, WCl'e periect, COII- sequently easily recognised. Some * I have obtained excellent examples of this fungus (MucorJ on the InteitlnaJ mucous membrane of the pig also, whilst subjecting strips of the intestine to OOntluuoiU übaervation. CYSTS &.. SIMULATING HALLIERS CHOLERA FUNGUS 1,1 ,1 L.OPI 1) IN ORDINARY EVACUATION THE CHOLERA FUNGUS OF TITOME* ANT) OF HALLIER 16 appearance. The cells had become nearly everywhere strung together, and long filaments of oidium Thom<." cholera fungus " of lactia (Fig. xxiii), corresponding ex- actly to the figures given by Thome of the cholera fungus discovered by him, to which rather a long name was given at the time, viz., " Cylindroicenium Cholera Asiatics" This condition lasted till the sixth day, when a crop of a white mould was perceptible in the isolated preparation, and a plentiful crop of penicillium and aspergillus appeared on the other cultivation (Fig. xxii). This slide having become rather dry, a few drops of distilled water were after this occasionally added. On the eighth day long delicate filaments were seen growing out of the condition of the prepara- white humus-looking substance in tion under bell-glass after ,-• , -iji jjii first week. the apparatus, and on the tenth day other filaments were observed, which seemed to be tipped with various coloured heads, apparently of the same kind as on the other slide, those of a bluish and yellowish-brown tint prevailing; but by the eighteenth day the long delicate filaments had grown over them, the whole surface of the preparation presenting a woolly appearance. After this no further change could be seen to take place in either cultivation, The apparatus opened on i j.i p j. ± fl j. ] f xi the twenty-first day. ail(i 011 Tlie tWCnTy-niht Clay 01 tlie experiment the bell-glass was opened, and the glass plate placed on the stage of the microscope. Precisely the same species of aspcrr/Ulus and penicillium were found as existed in the non-isolated cultivations, with the addition that great numbers of the filaments forming the white flocculent tuft bore at their terminations cysts or sporangia filled with distinct spores (Fig. xxiv, 1-4), which, I think, correspond exactly to the Cy »\ ob i tained ?» ct i y lik I cysts figured by Professor Hallier the "cholera-cysts" figured v. i • j i i by Haiiier. oi the immature cholera-cysts, whose drawing has already been given and may be compared with this.* Aspergillus tufts were present in great numbers : nearly all of them had fallen off from a germinating aspergillus tl j filaments among the mycelium; tuft simulating a "cholera o J cyst" in the same condition: a ICW, IIOWeVCr, Were perfect, COU- sequently easily recognised. Some * I liave obtained excellent examples of this fungus (MucorJ on the intestinal mucous membrane of the pig also, whilst subjecting strips of the intestine to continuous observation. 17 DEVELOPED IN ORDINARY STOOL. of these fallen masses were germinating (Fig. xxvii, 2), and presented, as nearly as anything possibly could, the appearance of the mass of spores figured by Professor Ilallier as a "cholera-cyst" in process of germination (Fig. i, 11). As the preparation was dry, a few of the cysts were transferred to another slide and water added, upon which many of the capsules of the sporangia gradually ruptured, and the spores escaped (Fig. xxv), a bare columella and the ruin of the capsule alone remaining. As it was not advisable to expose the preparation during the experiment, the various stages in the development of these cysts were not followed, in order to ascertain which some spores and cysts were sown on the juice of various fruits, boiled and unboiled, and on pieces of cheese. They rapidly germinated, and in those preparations which were sown in cells on the slide without a covering glass, produced precisely similar cysts to those sown; when, however, covering glasses were used, the fructification was not so perfect. For example, a glass slide was taken, and two semi-circles of asphalt varnish were brushed on it, one being rather larger than the other, so that the ends of one half-circle might overlap the other, but not so closely as not to permit the entrance and exit of air, as may be learnt from the Figure (xxix). When nearly dry, a minute quantity of growing fluid, consisting of a solution of grapesugar and phosphate of ammonia, was placed in the centre, upon which a few spores were sown, The cysts and spores sown a t i lin covermi? glass being placed on a slide, and the develop- . -i? -i 1 ment described step by step. OVCr it, wllicll adhered to tllO SCmi- dried varnish. The slide was placed under a bell-glass, kept damp by being lined with some moist blotting-paper, at an average temperature of 90° Fahr. In the course of six hours a clear oil-like spot appeared in the spores, and on the second day they were germinating rapidly (Fig. xxviii, 1). On the third day tho field was crowded with mycelial filaments (Fig. xxviii, 2), and on the seventh day a filament which had crept beyond the droplet of fluid into the free space between it and the varnish bore a distinct sporangium (Fig. xxviii, 3). Separate spores, however, were not distinguishable in this cyst. These illustrations will, I think, be sufficient evidence to show — (1) that the cholera-cysts figured by Professor Ilallier are not always Deductions. obtainable from choleraic discharges, (2) not confined to 18 CORPUSCLES TIESEMBLIiVG SPORES. cholera, (3) nor even to diseased conditions of the intestine, but (1) may be cultivated from the stool of perfectly healthy persons. The experiments instituted to test t\id observation as to the inoculability of rice plants Inoou lability ol" rice plants. -. , -, " ? „ x have as yet not been satisfactory , conti — fj * — sequently no conclusions have been arrived at on the matter. SECTION II.—" SPORES." It is by no menus so easy to explain what the yellowish more or less oval hyaline bodies are which Professor Hallier calls "spores" (vide Fig. i, 3) ; such bodies are exceedingly common in choleraic discharges, and I believe are very different in their nature ; but whether corpuscles simulating the any of them arc " spores" will, I byHam e r en ° f " SPOVCH " think, be satisfactorily explained in the sequel. The objects I have met with in cholera discharges more or less resembling these bodies may be arranged into four classes : — (1). — Globules of a fatty nature ; (2). — Altered blood-cells ; (3). — Corpuscles embedded in the tenacious substance coin- posing the "flakes;" and, (4). — Globular conditions of certain infusoria. 1. Persons accustomed to microscopic work must have found that to distinguish fat or oil globules from other bodies very different in their nature, is not always so easy a matter as is commonly stated in text books on the subject. It lias frequently occurred during this investigation that, in spite of the addition of heat, absolute The frequent difficulty of alcohol, rectified ether, potash, iodine 2SSXSS ££?&*?*£ ™ d °®r re-agents, not overlooking buies. the prolonged application of carmine, I have failed in distinguishing with certainty fat globules from pellets of slimy substances endowed with life, when both were known to be present. Indeed, I have frequently mixed fat with gum water and other substances for the purpose of testing the value of the re-agents which had been applied to ¦kxlies under examination, and have found that, in a great number of instances, the results are fallacious ; either the globules remain unaltered, or both FIG. XXVIII. FUNGUS VERY LIKE HALLI ERS CHOLERA FU NGU ntvnnfFn in thf r,pnwi Nn- si lOF XXIX. FROM ORDINA kinds are destroyed, or they are acted upon indiscriminately. A fair sample of this difficulty is carefully delineated at Figure xxx, representing objects very like delicate " cysts " and " spores," which being watched for eight hours remained unaltered, resisting pressure, &c, but broke down in twentyfour hours into unmistakable globules of oil. Having experienced very great difficulty in this matter, I propose giving one more example of a condition which is a particularly prominent feature in the early stools of a cholera patient ; indeed, for a long time I was unable to persuade myself that it was not a condition of some low form of life, especially when the globules were highly coloured, or when the homogeneous contents of the pellicle shifted its position. A sailor was admitted into hospital with all the symptoms of cholera, and, at the time this evacuation was obtained, suffered from severe cramps. The stool was examined three minutes after being voided, was found to be alkaline and of a muddy colour. The sediment consisted almost entirely of greenish-yellow corpuscles, varying considerably in size, the larger ones being flattened out under the covering glass (Fig. xxxi) ; many of these having the contents contracted, the contour of a delicate, filmy capsule being evident at the spot Jo^To^ZtynZ:* by where shrinking seemed to have taken place (Fig. xxxii). They were gene- rally spherical (1), but many were oval (2), and a few were seen presenting several hyaline projections whilst rolling in the fluid on the slide (3). In some cases they retained their form and appearance for a long time, but the greater number lasted only for a few hours. They were frequently observed to vanish suddenly like a distended blood- cell, leaving only a ring behind (Fig. xxxiii, 1), previous to which, in a few instances, a slightly granular appearance was presented (2J,aiid the ring was often seen particularly granular (3, 4, 5), as if all the contained granules had adhered to it. The globule in the centre of the figure, with the contents separated from its enclosing pellicle (6), was watched for a long time, but no alteration in its appearance occurred. Other similar bodies were watched continuously for three hours with the same result, save that they gradually became excessively transparent, visible only by careful adjustment of the mirror. In the course of four or five hours the entire field presented the appearance delineated in the figure last alluded to. At Figure xxxiv a regular colony is seen 19 20 DISTENSION OF SI'OIIES BY FLUID. of those globules surrounding 1 a crystal. They also disappeared, in the course of a few hours. Rectified ether caused the pellicle to present a minute granular appearance, and those which had the contents puckered became Bii'cct of re-agents. symmetrical. Boiling in ether seemed to thicken the pellicle. A portion of this was set aside until the next day, and was found to have retained its condition, whereas the globules in the evacuation set aside in the vial had disappeared. Absolute alcohol subsequently added to the boiled portion seemed rather to diminish their number. In some cases one globule was observed to " melt " into the other, so as to form one globule ; otherwise no change was observable. Solution of chloride of zinc and iodine, — some became shrunken and irregular, others continued spherical, but with a finely granular pellicle. Solution of iodine only caused several of them to become very transparent — scarcely visible, were it not for the slight tint communicated to them. Liquor potasxee causes them to lose their yellow colour ; they become perfectly transparent, except that a few molecules which existed within are brought to view. A few of the globules Avithstand the re-agent for some time. Acetic acid seemed to coagulate the pellicle, as it became finely granular : very much the same appearance as followed the addition of alcohol. Dilute sulphuric acid caused the contents to contract, but the colour was retained, or it became slightly brown. Dilute nitric and hydrochloric acids acted in the same way. I have made many attempts artificially to produce globules of this kind, the nearest Their artificial production. *"* -. , .' p ? , approach being a mixture ol melted butter, albumen, and gum water well shaken together, and at the time of examination adding a little thick syrup so as to cause the puckering to take place between the pellicle and the contained fat. The action of re-agents, however, on this pellicle was slightly different to the foregoing. Bpoi*es immersed in fluids of varying density become greatly altered in their appearance; frequently the outer layer becomes so attenuated, and perhaps stained, that it is a matter of great difficulty to state of spores by positively that the cell pellicle sur- rounding the protoplasm of a spore differs from the clearly defined outline of a globule of oil, in spite of a knowledge of the action of re-agents, and of the varying powers of refraction which liquids manifest. Hence it is not impossible, nor inexcusable, that Professor Hallier in some instances might have been deceived by these ajypcarances, especially as it is evident from the conclusions he draws concerning the importance of some of the " cysts" in M. Robin's plate (which are undoubtedly fat), that he had not made prolonged microscopic examinations of ordinary excreta : the Professor, however, had more spore-like objects to deal with than fat, such as the ones described in the next and following paragraphs. 2. Almost invariably circular cells are observed in choleraic dejections of a errccmsh- Altered blood-corpuscles. -.-, ,° . , .. . D . yellow or brownish tint; contents generally homogeneous, and the capsules very delicate. The microscopic appearance of one of these capsules is here represented at different distances from the objectglass, the size selected being about the average (Fig. xxxv). The appearance of the capsule a little before the focus is attained is shown at 1, a clear spot shading off into a dark ring. On bringing the object-glass nearer to it, the defined outline of a spherical body is £ZL£5 -ESSS" seen with slight opacity in tho centre (2) ; and on attaining the exact focus, a greenish-yellow perfectly hyaline sphere is brought to view (3) . On going beyond this, a dark spot is seen in the centre, gradually shading off towards the periphery (4) ; when the light is shut off almost entirely, a slightly irregular space is seen presenting a very slight pink tint (5) ; this particular cell was constantly watched for three hours, when suddenly it became transparent, and required most careful illumination and focusing to make it visible, a delicate ring of a slightly diminished diameter being all that remained (6). These, however are not always spherical ; frequently a very filmy tongue-like projection is observed (Pig. xxxvi, 1), sometimes more than one (2) ; it is projected exceedingly slowly, and then retracted amceba- One or more vesicle-like i'i r/ , a i?j., v ,- _ x'^^ j_i • j.* protrusions. llke - Alter a time this action ceases, the projected vesicle-like tongue is either permanently retracted, or is left out rolling about with the corpuscle in the fluid (3). These arc doubtless distended 22 EMBRYOS OF A ROUND- WORM IN " CIIYLOUS URINE." blood-cells, a great number of which may exist without yielding the slightest trace of colour to a rice-water evacuation. Whilst following the changes taking place in these particular corpuscles in various fluids, I had opportunities of making an examination of the urine of a patient in the General Hospital under the care of Dr. Lyons, who had been suffering from the condition Precisely similar appear- kllOWn as " CkljloilS IM'tiie" for about ances observed in a case of ,¦¦ , ,-. ? 1 .... « chgiom urine." a month, together with pam m the right testicle, and great emaciation, in spite of good food and a good appetite. As the colour so closely resembled many rice-water stools, I carefully examined it, and was repaid in a way I had not anticipated. It was albuminous to the extent of about one-fourth of its bulk, slightly acid, with a specific gravity of T015 ; ether caused a separation into two layers, a clear urine-like fluid containing oil molecules, and a white homogeneous mass consisting of minutely molecular debris. Before the addition of re-agents the fluid under the microscope so closely resembled the condition of a cholera stool just described, as not to be distinguishable from it ; yellowish-green cells, some hyaline, some granular, some protruding a tongue-like prominence?, and others with the contained plasma puckered in various ways (i"ig. xxxvii). A few of the larger corpuscles were seen to shift themselves (like an amoeba) a distance fully their own diameter, the shape altering at the same time. At first I doubted that they really were blood-cells, as the extent of variation in size was considerable, as shown by reference to the figure, which is carefully drawn to scale. The fluid very quickly gelatinised in the test tube ; indeed it frequently does so in the Gelatinisation of the urine. j • j_ > i i i t • • • j. i patient's bladder, giving rise to stop- pages during micturition. I have not seen cholera discharges spontaneously gelatinise, although such a condition is said to occur. A portion of the coagulated mass (which when stirred closely resembled a lump of moist gluten) was teased on a slide with needles and examined. It consisted of fibrillse studded Avith blood; granular cells, scarcely differing from those seen in cholera discharge Hakes, Appearance of coagulum. , , 1 except, perhaps, in being more uni- versally granular. They seemed to present more of the character of pus-cells. In the midst of this fibro-albuminous matter several embt'l/oi> of a lloaud-toona were discovered every time the urine FIGS. XXXV XXXIX THE CORPUSCLES IMBEDDED EN THE FLOCCULI. 23 was examined, one of which is seen coiled up in the drawing (Fig. xxxviii). A careful sketch of a larger one, after the addition of acetic acid, is given at The embryos of a round- XXxix. In tllC COUrSC of a few worm detected. p , ? .. . , - minutes, when the sketch was nearly completed, a caudal-bursa became visible under the influence of the acid, and is delineated at No. 2. When first seen, I thought they were some detached filaments of a fungus, judging from the hyaline, structureless appearance presented ; after a time, however, a few of them were observed to move very Approximation of diameter. nature was at an end. It will not be surprising that the existence of these was not suspected, when we consider that fully two hundred of the larger size figured could pass abreast through a very small pin-hole, an orifice not exceeding the fiftieth of an inch in diameter, as may be verified by a simple calculation. Perhaps this fact may help to throw some light on a very obscure disease, of which little is known beyond the symptoms, although frequently met with in some parts of the world ; and, indeed, may perhaps account for its localisation to such places as the West Coast of Africa, where I am told it is by no means a rare malady. As the mature worm still retains a hold on its victim, being perhaps safely lodged in the kidney, and not having seen an embryo of this kind before, nor yet a drawing, I must leave to a more experienced helminthologist to decide to what species of nematode it belongs.* 3. In examination of this class of corpuscle, namely, those intimately associated with the The corpuscles associated well-known flakes in cholera deiccwith the "flakes" change in , . ? . « ? , , . . ° appearance very quickly. tions, it is of the greatest importance that the evacuation should be a recent one, because its character may be entirely changed in the course of an hour or two. Sometimes, however, the change is not so rapid, depending on the chemical nature of the fluid, especially od the extent of its alkalinity — cholera stools being almost invariably alkaline. The method adopted in these examinations is to pour the discharge into a conical * While tills report was passing through the press, the " chylous" condition which this urine had presented for more than two months gradually disappeared, and so did all traces of albumen, and of the embryo-worms. 24 THE CORPUSCLES IMBEDDED IN THE FLOCCULI. l vessel, set it aside for a short time, and, when the sediment is seen to have been deposited, a pipette is introduced in order to transfer a portion of it to the slide. Frequently the sediment is seen to he of a very slimy nature, requiring some tact in bringing it into the pipette. Illustration I: — The evacuation was from a man suffering for eight hours from a severe form of cholera, who died on the second day. It was of a pale straw colour, with a muco-flocculent deposit. In the upper liquid por- case exemplifying these tion nothing special was visible, but corpuscles in an early condi- cxammation of Rediment ft was found to consist of flakes of a gelatinous semi-fibrous texture, studded with globules, circular and oval, with a pale yellow tint, and of a homogeneous nature, a very correct representation of which is given at Figure xl. In some of these bodies a clear space is observed, but nothing further could be made of their nature. lodine stained some of a brownish-red, and others of a deep yellow. Liquor potassce seemed to make the corpuscles more distinct at first, and to isolate the contained granules and molecules, Effect of re-agents. giving the contents a distinctly dotted appearance. The fibrillatcd substance became slightly granular, then it gradually faded, and so did the corpuscles, which in the course of half an hour entirely disappeared, except here and there a little cluster of molecules, five or six, with a clear space in the centre, all trace of the fibrillated texture having disappeared. Acetic acid increases the stringy appearance at first, making each little fibril appear dotted, like a very fine bead of granules, or minute molecules; eventually the fibrillated appearance is obliterated altogether, a diffused, finely granular substance being universal. The corpuscles maintain a sharply defined outline ; the continuity of the outline, however, seems frequently somewhat broken in one or two places, as if the circle were formed of two or three short vibriones imperfectly united at their ends. The next day the sediment was still slimy, and could not be taken up by means of a delicately pointed pipette. It still consisted of a FIGS. XL XLV FLOCCULI &c. IN CHOLERA STOOL streaky, semi-membranaccous substance, but the imbedded cells bad cither become transparent, The appearance on the ()1> presented a granular or minutely second *. ,_ /ti' ?i.-\ , *j i. molecular appearance (Fig. xli), with no distinct cell wall. Solution of chloride of gold picked I them out very distinctly. On the third day the flakes had lost their membranaoeous character altogether, but many of the granular corpuscles remained. On the and succeeding days. fourth day a few animalcule were seen, which enormously increased by the fifth (Pig. xlii). On three occasions only have I observed the appearance of this protozoon in choleraic discharges. Several evacuations from the same patient were subsequently examined; the flakes, however, did not present the fat globule-like appearance again, but molecular, as shown in { the previous Figure (xli). II : — Another case, the third liquid stool, presented a yellow colour, about one-sixth of which was Case in which the corpus- COITipOSCd of a whitish floCClllont SC(lik cles were granular when the niCnt, presenting precisely tllC SaiUC tIST IZZ^T^t microscopical character as the second iments. stage of the last ¦ described ; a semi( membranaceous substance, dotted With irregularly defined cells (Pig. xliii), very like what is seen in exudations effused in catarrh. On very careful watching they arc seen to protrude excessively delicate processes of an amoeboid character (Eig. xliv), just as the white bloodcorpuscles do. liquor potasscp- caused the membranaceous appearance to vanish after a time, reducing the cells to an aggregation of granular or Effect of re-agents. tolecular particles, ffiher docs not destroy them, nor does etic acid, but it seemed to make manifest a delicate cell '-wall ; and iodine superadded enhanced this appearance, in Jlnany cases causing the contents to collect at one part of the cell (Fig. xlv). The membranaceous appearance had disappeared in the fluid on the fourth day, but the granular cells remained visible Jbr nearly a week. n 25 26 THE " STILL" CONDITION OF ANIMALCULE. \ Illustration III ;- The fifth evacuation of a patient suffering from the cold stage of cholera was examined half an Case in which hyaline and hour alter it Was passed. It was SSSSL bodi6S ar6 S6en colourless, with a few shreddy flocculi floating in it. It was slightly alkaline. The flakes presented the same membranaceous appearance as in the foregoing example (Mg. xliii), with numerous corpuscles, more or less intimately held in the meshes of this texture, a great number, however, being dispersed in the fluid ; some were oil-like and some granular, examples of both kinds being spherical and oval, and the gradations from the merest particle of slimy or oily matter to the complete corpuscle were so fine, that it was impossible to point out any salient distinguishing character about them. When free, the hyaline and granular corpuscles were more or less round, but when contained in the meshes of this fibrillated texture, were generally elongated, as shown in the drawing (Fig. xlvi). lodine solution being added to the slide, it was observed that whereas some of them were coloured brownish-red, the greater Effect of iodine, portion became merely stained by the ordinary tint of the iodine (Kg. xlvii) ; all, however, in the course of the day becoming granular, but the distinction of brown-red and mere yellow remained. In the course of an hour other slides were prepared, but the microscopic appearance had become totally different. The oil-like bodies, of whatever shape, had become granular, and the iield presented exactly the same appearance as presented in Figures xli and xliii, while the addition of reagents produced the same results. On the fourth day all traces of corpuscles had passed away, merely broken doAvn molecular matter remaining. 4. Intermixed with the corpuscles already described are others to which I wish to allude The "still" circular condi- '>¦% ±r\ x i- i.' ~i~? xi tion of animalcule with the greatest caution. Frequently a globule has been observed for some time, and finally disposed of as being merely an oil one, when suddenly it is seen to protrude a portion of its substance; retract it, and while so doing another protrusion FIGS. XLVI LI. VARIOUS OBJECTS IN CHOLERA STOOL 27 ACTIVE CONDITION OF THE ANIMALCULE. becomes visible at some other portion of the little mass, and then, perhaps, it will shift its position, exactly after the manner of an amoeba. These are frequently hyaline in a fresh stool, but generally granular ; no trace of nucleus or contractile vesicle can be observed ; sometimes they are very numerous, but when there are other corpuscles in the field which act in a some- what similar manner, it is impossible B e A »rr»n y Tr»ui« yi "'"' bUt to «V . to class they belong, unless, indeed, they move across the 7 ¦> fj field like an ordinary amoeba, and not merely content themselves with protruding portions of their substance into the surrounding fluid, as was stated the corpuscles in the last described kind did. I am not in a position to state that these are the "still" and amoeboid conditions of more than one kind of animalcuke ; probably they are, but that they are so of one kind, I think I may state pretty definitely ; and, as they are sometimes distinguishable in the still globular condition for a considerable time, they really may have been the bodies seen by Professor Hallier, and mistaken -r. "ut* *^ ¦ by him for swollen spores; most Possibility of their having J - J- ' , been mistaken for swollen frequently, however, they are oi short spores. dnvn+irvn duration. The cause of this variableness I am not in a position to state. These bodies were noticed very early in the course of the inquiry, and every particular concerning them noted ; but I have to confess that not a few links are wanting in the "life history" of these animalcules, which the following illustrations will but too plainly demonstrate. Illustration I: — A pale, straw-coloured, perfectly liquid stool, in which the sediment was very scanty, was obtained from a patient in the cold stage of cholera. The dejections being passed involuntarily, numerous little heaps of sarcince were present (Pig. xlviii), as indeed exist to a Presence of sarcinae almost „„ i p „ ]„ p Y f Pll f jv, riP'll'lv nil tllP universal in cholera stools. gieatoi oi less extent m ne.u ly an me cholera evacuations examined, with numerous masses of a granular or jelly-like substance, in which yellow translucent lumps arc imbedded, probably of a fatty nature (Pig. xlix, 1) ; together with masses of a someAvhat similar outline observed to alter in form very slowly, as at 2. In some cases a pellicle becomes evident, when the contained TRANSITION FROM THE ACTIVE TO THE " STILL" CONDITION. 28 jelly-like protoplasm contracts, as at 3, the various forms assumed by which arc represented at Alterations in the form of Figure 1, with a great number of more the spherical bodies very gpherical bodies VCTV like oil gradual. 1 ?. ?\ J globules (Figure li) ; some arc seen to be flattened out (1), others protruding a vesicle exceedingly slow iy ; the body at No, 2 becoming in the course of five minutes to the condition delineated at 3, 4, 5; whilst great numbers of a minute animalcule were Presence of exceedingly geen ac tively HlOviag amOlli* 1 them all ; •live animalculsc ; ? * n H-. . ° sometimes one flagellum is seen v pos- terior one, at others an anterior one also, both being retractile at will, and another may be darted forth out of any portion of its body. No organized structure can be seen, neither mouth nor eye spot, nor any trace of contractile vesicle, merely a spindle-shaped speck of jelly enclosed structureless; perpetually j^ delicate elastic SaC, endowed with (.¦hanging in lorni. ? . , the power of rapidly altering its shape and position (Fig. In). So capable are they of adapting themselves to circumstances as to be able to insinuate themselves with the fluid through the meshes of fine blottingpaper. All these were present to a greater or less extent for a week. A drop of the fluid was placed on a hermetically sealed slide, and the little bodies remained active until the fourth day, when they gradually ceased to present any kind of motion, but settled down into irre- ooTdS:n a ° tive Sul" little masses of jelly-like appear- ance, to which condition also the cor- puscular bodies had been reduced (Fig. liii). On several occasions, however, the animalcuke were seen to become more than usually active for a short time, before ceasing altogether : to push out processes in all directions, and as quickly taking them in again, finally settling down as shapeless little pellets. Some of the various forms assumed by one of these at this stage are sketched in Figure liv. Illustration II: — A condition precisely similar to the foregoing was observed in the evacuation of another man a i'ow hours before death, as well as in the contents of the large and small intestine at the post-mortem examination. The action of re-agents is much the same as on any other hyaline protein "lobule. FIGS. LI! LVII. STAGES IN THE EXISTENCE OF THE AN IMALCU LE AND THE REV D MRM R BERKELEY'S GROWING CELL They remained unaffected by strong acetic acid for ten minutes ; gradually, however, the contents contracted more or less regu- Action of re-agents. larly, thus allowing of a delicate capsule being brought to view (Fig. lv, 1). After a time the contents vanished, merely a finely granular ring being left (2) ; absolute alcohol made the contents appear granular, as at 3, whilst some appeared but little affected ; ether subsequently being added caused them to shrink considerably, but did not dissolve them (10. lodine stains them a brownish-red and makes them appear somewhat granular (5). In order to test whether some of them might not be " spores," a series of observations was commenced, some of which have already been described in the chapter on " Cysts" (page 11). A growing-cell was prepared on the Hevd. Mr. Berkeley's plan, by drawing a ring of varnish on the glass slide, allow- ing it to become nearly dry, cleansing Jte Berkeley's growing. ag the covering glass, tho- roughly with spirit and distilled water. A droplet of the evacuation was then transferred to the centre of the cell (Pig. lvi), care having been taken that no part of the sides was touched by the fluid when the covering glass was applied. It was afterwards hermetically sealed, sufficient air being already enclosed to allow at least of germination. The limited area of the preparation enabled the geography of various objects to be easily remembered, and tended very materially to precise observation. On the second day the corpuscular bodies appeared to be more granular or less like oil globules, frequently with one or more indistinctly visible vacuola) (Eig. lvii, 1) ; many are elongated and presenting very slight movements (lvii, 2). A few animalcules were still present; germinating spores were also visible. On the third day the circular and oval bodies had almost entirely disappeared, but on approach- Transition of the "still" to ing the margin of the fluid immense the active condition on the -t t> ,-> • i i ± j.i third day. numbers ot the animalcule, to the extent of half the field of the micros- cope, were seen moving about with great rapidity and perpetually altering their form, a clear space being observed iti some of them (Eig. lvii, 3). 29 BLOOD-CELLS MIXED WITH AM(EBOID CORPUSCLES. 30 On the fourth day the activity of the little animalculse had diminished, many were gradually re-assuming the circular condition. Thinking that this was Return to the "still" con. &n i nc li ca tioil for a fl'Csll supply of ail 1 , dition. ? . , , i t n the varnish was scratched away from a small portion of the side with a needle, watching the effect under the microscope while doing so. They did not appear to be particularly affected by this proceeding, for in the course of an hour they had all become circular, and almost motionless ; many attempts were made to get at a more complete life history than this, but hitherto without success. The duration of the corpuscles and of the active ani- malculse is very variable, sometimes After becoming dried may eas ily rCCOJmizcd in stools which be revived by the addition of , •', , ° , ? ? ? fiuid# have been kept lor a month ; on other occasions disappear in a few hours. They have frequently been seen after having been thoroughly dried to re-assume active movements on the addition of fluid ; but exposure to the sun at a temperature of 120° Fahr. stops all movements, no matter in what fluid they are placed, becoming sometimes completely disintegrated, but they will re-appear in such a fluid after a time under favourable circumstances — probably new ones being developed. These bodies are not confined to any particular stage of cholera, as the following will prove. Illustration 111: — The dejection of a person, shortly after the first symptoms of cholera set in, was obtained for examination. It was about the third liquid stool, of a pale yellow colour, slightly alkaline to test-paper, with the average amount of sediment. This consisted almost circular corpuscles present- entirely of the bodies sketched in %£E2LSS*££ lfi S urcs Ivfli to lx. First, a number cells and animalculce. of large granular cells, very delicate filmy spheres, rolling about under the covering glass (Fig. lviii) frequently, as if undergoing the process of division; secondly, corpuscles of the same granular appearance, but generally somewhat smaller, from which filmy vesicle-like projections were seen to proceed very, very slowly, and as slowly retracted, followed by a similar protrusion from another portion of its substance (Fig. lix, 1) } or two or more may be seen at FIGS.LVIII LX. EXAMINATION OF CHOLERA STOOL LXI „ HEALTHY BLOOD IN DITTO LXII LXIII „ „ ORDINARY STOOL the same time (2). The granular and minutely molecular matter did not enter into these saccules, and I am not certain whether an inner or an outer wall exists, but sometimes it seemed very like as if the outer gave way for a filmy inner lining to come forth (3), at other times it seemed quite the reverse (4). After a short time the projections in many cases appeared no longer to be retracted, and were seen to curve upon the cell as the evaporating fluid bore it along (5). Some, however, are seen to be of larger size (G). Thirdly, blood-cells which have assumed very peculiar outlines, the result of diosmosis (7) ; and fourthly, innumerable animalculse, of the kind already alluded to, exhibiting great activity amongst the various cells in the field (8). The patient died on the next day, but a stool was exa- mined a few minutes before death ; s ti^fo C ;tod-c a e C iS nStliem - was highly coloured with blood and contained a great number of animalculae (Pig. lx). Some of these were tugging at the blood-cells and altering their form, distinctly pulling the pellicle or cell envelope away from the enclosed plasma.. No. 1 was altered to 2, 3, and 4 in the manner described and shown in the figure. The animalculse presented an unusual appearance ; either a large clear space existed in most of them, of the same size as the blood-cells, or one or two blood-cells had become engulphed in their homogeneous substance. In some cases they were distinctly seen to bo merely adherent, the little creature rushing along as if it had no burden. The next day the blood-cells had become granular, but the animalcula) were as plentiful as ever, and continued so for a fortnight, everything else having broken down. Having now given a brief account of these few classes of corpuscular bodies, and shown that Are any of these corpus- none o f them were seen to germinate cular bodies peculiar to cho- ?, ? „„ . . ° lerap like the spores of fungi, the question naturally arises — Are any of them peculiar to cholera ? The first class, namely, those of a fatty nature, need not be considered, for no one will The first and second classes of suppose them to be peculiar to the COipilSCieS. Hicoocn • ¦fno como mov \\n cciirl drv-n noim disease ; the same may be said concerning the presence of blood, and as to the shape assumed by its corpuscles, the figure already given in connection with " chylous urine" will show that there is nothing peculiar about it, nor yet about the amoebalike movements" of the blood-corpuscles, as the following easily repeated little experiment will show. A small portion of a slightly alkaline cholera evacuation was iiltcred off into a test tube, and having pricked my finger, a few drops of blood was allowed to fall into the fluid, with which it was immediately mixed, and a drop of the mixture transferred on a slide to the microscope ; nearly all the red cells were seen to present a stellate or ecchinnulate appear- ance (Fig. lxi, 1) ; only a few white Changes occurring in fresh- COrDUSCIcS WCI'C Visible, and tIICSO ly drawn healthy blood when i j 1 i placed in fluid. presented a granular, more or less circular outline (2). Some, however, were spread out like an amoeba (3), but no movements were seen. In the course of two hours the stellate form of the red cells had disappeared, and presented the various forms commonly seen in evacuations (4). Having been unable to sec any of the white cells protrude portions of their "substance, it occurred to me that, perhaps, the temperature of the fluid being only 80° was the cause; consequently another portion of the fluid was filtered and carefully warmed up to ] 10°, when a drop or two of freshly drawn blood was introduced. This time a very slightly granular white cell was seen to alter its form and protrude Warming the fluid favor- 0110 01' two Vesicles from its Substance able to the exhibition of amoa- (k\ «,-,,] /l vnw tliom in no>nin whi'nh bold movements in the blood- \°)> a ™ 1 iilCi } V tJieill HI again, WHICH it continued to do for a few minutes, then ceased, becoming more granular cells. than it was before. Others were observed to act in the same way ; one pale white cell was seen to possess a very delicate filmy capsule, extending some distance beyond the contents (0); it suddenly vanished altogether, leaving a merely irregular granular heap to mark its position. The four tli class (it will be more convenient to consider the third afterwards), namely, the The fourth class of corpus- var ious stages of the animalcule, was Cl©S« n • i 111* . -a for a considerable time the subject of much curiosity, especially the kind described as presenting such activity. The fact of their being almost universally present in choleraic dejecta, and yet never, as far as I know, 32 alluded to, except indeed that Thiersch of Erlangen could have seen one of these on the point of passing into the " still" condition, during which stage pseudopoda are incessantly projected in all directions, when he Aetmophrys-like stage. t n i i i , • speaks of having observed actmophrys-like bodies in some choleraic dejecta which he had examined, and wondered what they were.* There was some difficulty in tracing this body to any of the described sjoecies of animalcule©. Its minute and rapid motion added to the difficulty, as well as the variableness of its shape, because although generally spindle-shaped, it may become round, triangular, or stellate in less than a second ; frequently a succession of pseudopoda arc seen projected in a wave-like manner, as if lashing the fluid when about to pass out of the active state. It is generally hyaline, but may be granular; sometimes a vacuole is observed, but a contractile one never. There is always a very delicate posterior filament, at first continuous with the sarcode, and a still more delicate anterior one, both retractile. In some respects it agrees with the description of the Monad Bodo t but as Cienkowski, in his celebrated article in Solmlze's " Archiv," distinctly states that in the amcebiform stage of all the true monads the pseudopoda are pointed, whilst in the amoebiform stage of this animalcule the projections are, I think, invariably rounded, so that for this and other reasons, which need not be entered into here, room may probably bo found for them among probable family to which the Astasisea or Euglenaea family, so the animalcule belongs, and • , \ m i its connection with water. common in our tanks. Theassocia- tion of a cholera entozoon with the c " one species of which, when in its mature condition, causes the red colour observed in so many pools, and which Ehrenberg thought was the means by which the miracle was brought about of turning the waters of Egypt into blood, — the finding of precisely similar animalcule in drains, gave rise, as may be supposed, to not a few very pretty theories, which, I regret to say, like many others, had to be abandoned altogether. A gentleman, with whose personal habits I am well acquainted, suddenly felt some griping pains with inclination to go to stool, but was otherwise perfectly healthy. The * The animalcules alluded to in this Report do not in any wny resemble the figures of the actinophrys-like protozoa accompanying Dr. Sanderson's account of his celebrated experiments published in Mr. Simon's Ninth Report. a 33 34 THE ANIMALCUL.E FOUND IN ORDINARY EVACUATIONS. motion was very scanty and very diluted, but was followed immediate relief. It occurred to iJaTooi? other - than cho " me to subject the stool to a micros- copic examination, and, to my sur- prise, these animalcule, both in the active and " still" stages, were present in the most perfect condition, together with numerous globules of a fatty nature, exactly similar to those already alluded to. A comparison of the figures here given (Figs, lxii — iv) with ones previously described will, I think, be sufficient without repeating that description. The next stool passed by this individual was also a relaxed one, and microscopically of the same character, after which the motions were perfectly natural; but, in proportion as the motions became more solid, the ease with which these animalcuke could be found diminished. Many other ordinary evacuations were examined, and in fully half, after more or less careful search, they were discovered. After a brisk purgative they are frequently seen in great perfection. In alluding to the nature of the third class of bodies, namely, those found in the meshes The third class of corpuscles. „,, **.-,-. ?, . -¦ ¦¦ . oi the nbrillated substance composing the flakes in cholera evacuation, I wish to premise that the remarks are reservedly made, as the subject belongs more directly to the pathological anatomy of cholera, which subject forms a later part of the programme drawn out for guidance in connection with this inquiry. It will, of course, be understood that the corpuscles of the former three classes arc also found with the corpuscles forming this division; indeed, it is frequently impossible to Difficulty of distinguish, separate them, especially from those XfoyLSoXTe^e amcebiform conditions of animalcule which are seen so frequently in evac- uations. This is probably the reason why so many different descriptions exist of their appearance and of the action of re-agents. Now, the chief statement I have to make concerning the corpuscles of this class is, that they exhibit movements someivhat Mice the movements associated luith the amoeba. This fact may, by very careful these corpuscles exhibit some- cxammfl tj on W ith a i?OOd \of an what similar movements. . . , , S 8 wx ctxJ - inch object-glass, be verified by any A "BSE^i OE OF lijxlL 11 1^1 j 1 \J IVI I^J IvlCE™ vv .A-TEX^ SPOOLS • 35 one accustomed to the use of the microscope in most cholera stools when perfectly fresh. A portion of the substance of the corpuscle is seen to creep out insensibly from the mass, and as insensibly return : unless the eye is carefully fixed on the body, and is already a more or less educated eye, the phenomenon is not detected, and the observer enters it as " disintegrated epithelium" in his note-book. It may perhaps be remarked that no draw- Absence of epithelium in j of columnar epithelium, said to cholera stools. , ° . . . r » be so universal in cholera dejecta, appears in this report. The reason is that its presence, to an appreciable extent, has not been observed in the contents of the intestines discharged during life ; indeed, the only occasions on which I have been able to observe it quite distinctly were in discharges voided a few minutes before death, a long interval having elapsed since the occurrence of a previous stool. It was Bochm, I think, who first laid great stress on the fact of the shedding of the epithelium in cholera about 1832, since which shoddmg of columnar epi- period it has been the general opinion by e ßoenm m ' inSlifGadVanCed in Germany, with the exception of Virchow and a few others. In the well known Bavarian report of 1857 I find great prominence given to this view, modified, however, Supported by many German | )y 1C remar k that, as a rule, Only tllC writers I*l i »j-it n ntn t 1/ ,—.__-____, broken down epithelium, or rather freed nuclei of such, arc seen. Dr. Beale also lays great stress on the diseased condition of the epithe- and by Beale and Macna- an( J latest authority Oil mara, ? _ ___v. , t^_. t, r _ _ »_Si__ the subject, Dr. Macnamara, follows Dr. Beale ; indeed, it is evident that Dr. Macnamara's explanation of many of the phenomena observed in this disease is based upon a conviction of the correctness of the views advanced by these writers. It is of the utmost importance in matters of this kind, as was pointed out by Professor Parkes in 1848, not to confound the microscopical appearance of the rice-water stools but contradicted by Parkes, passed during life with that of the conwho maintains that it is only T , i? x i • j. j • 1 x • l z>j. present after death. tents of the intestine obtained after death. In a brochure which was pub- lished by him on this subject at the time I find stated: — "With regard even to the separation of the epithelium, although from the facility with which this structure is shed, even during ordinary healthy processes, it does appear probable, a priori, 36 DR. PARKES ON THE NATURE OF THE CORPUSCLES, that it would be largely thrown off in cholera, there is absolutely no proof that it is so thrown off until after the death of the patient. The stools contain none, or a quantity not more considerable than is present in common diarrhoea."* Judging from the cholera stools which have come under my observation in Calcutta, — several hundred specimens, — I believe that not more than two out of twenty slides will contain distinct traces of columnar epithelium. That these corpuscles are the remains of diseased epithelium may, I think, be disproved without any reference to post-mortem appearances, which I wish at present to avoid ; first, by the fact that, under favourable circumstances, they exhibit movements exactly analogous to those seen in the blood, pus, lymph, chyle, and the so-called "mucus" corpuscles. Secondly, cell formations and minute Reasons for believing that flocculi, microscopically identical with these corpuscles are not bro- t] frequently be observed ken down epithelium nor J l «/ in their freed nuclei. under other conditions, and irom sources where it would be difficult to account for their presence were they epithelium fragments, such as in the fluid obtained by pricking a blistered surface. Thirdly, that even where portions of columnar epithelium are seen they will, I believe, almost invariably exhibit, no matter how much broken down the cell appears, the delicate rim or basement membrane lining the free end of the cell, believed by some to be pores communicating with the cell. The presence of epithelial fragments, when not excessive, may be readily accounted for by the process of renewal which takes place in all cells. Dr. Sharpey writes :—": — " The particles of columnar epithelium are undoubtedly subject to shedding and renovation. According to Donders and Kolliker, the columnar cells on the villi appear occasionally to cast off parts from their upper ends, with subsequent reparation of the loss ; that is, a cell enlarging, and a second nucleus appearing ; the upper and broader part with its nucleus and much of the cell contents separates, and the lower remaining portion with its nucleus grows again to the natural size." And., fourthly, the epithelium thus discovered in the dejecta will remain for weeks unchanged in the fluid in which it was found, showing that the action of the liquid portion of the stool is not so destructive to it as would be inferred if the numberless corpuscles seen were the result of the dis- * The italics are mine. integration of epithelium which had been shed. I think there is no doubt but that these are the " peculiar corpus- cles" first described by Dr. Parkes, Probable identity of the with probably tllC circular, " still " corpuscles with those dcs- « mif liHon of flip nnimnlfnl r p nllmlorl eribed by Parkes as "pecu- COnCllllOn OI 1110 «HUmdIGUUIC aiIUCICCI liar bodies." to in tlllS report, tllC IIUCrOSCOpiC appearance and the action of re- agents coincide so entirely with the minute description given of them in the author's work. I am as yet not in a position to verify the author's belief that they arc confined to any particular stage of the disease. I hope, however, to obtain more exact data on the subject in my next report. With respect to the nature or origin of these corpuscles and the fibrillated substance in which they arc imbedded, I have not been able to disprove, nor in any way to modify, the views expressed by the writer at Their probable nature, a ii fi mf » wTiati lio rlvpw flffpn+irm in modification of fibrine. 1110 111110 WllOn 11G UIOW cIILOIIUOn IO them in the following extract, which may appropriately serve as the concluding sentence of this paragraph : — " It is in the highest degree probable that they owe their origin to effused blood-plasma, which assumes, with great rapidity a low, ill-defined, and non-progressive organisation." SECTION III.—" MICROCOCCUS." The term " micrococcus" {milcros small, and kohhos kernel) is now pretty generally adopt- th^t^mSotTs?^ 0 ed on the continent by the class of writers who advocate the pre-existence of a germ, in some shape or other, to every living thing, this germ, which may be infinitely minute, being called its " micrococcus ;" whereas another class of writers, very numerous now in England as well as on the continent, maintain that the pre-existence of a germ is not necessary to the development of living objects, providing certain atmospheric, chemical, physical, and other agencies are present; the nature of the object developed depending on the relative proportion of these agencies or "forces." In short, that life is a creature of circumstances, those circumstances being of an entirely physical nature. The question of the existence or non-existence of a "germ" being of such great importance in connection with epidemics and infectious diseases generally, and its investigation 37 38 THE MINUTE FORMS OF LIFE IN CHOLERA STOOLS. associated with so many difficulties, I should have preferred not alluding to the subject of this section at present, not having had time to accumulate sufficient material to enahle me even to obtain a clear idea as to what changes take place, much less to attempt passing any opinion Reasons for not deferring concerning tIIOSC changes. As, llOWthe consideration, of the sub- ? • 1 i 1 11 _?_i,i, i-1,,,i »,„ ject of this section. ever, it might be thought that no attention had been given to this por- tion of Ilallier's theory, — in some respects the most important, and certainly the most difficult to disprove, — a few illustrations will be given of what has been done in the matter. As already explained, the micrococcus, or germ of cholera, is, in the opinion of Hallier, the disin- Hallier maintains that the tcgrated SpOl'CS of a Special fungUS, Sra^p^aSeS:; which escaping into water may bo fungus. swallowed, or after being wafted by the air, adding a trifle to the "dust," according to Professor Tyndall, so prevalent therein, reach the interior of the human body, there to develop at the expense of the nitrogenous material, notably the epithelium of the intestinal canal. It will,, of course, be evident that the attempts, already described, to produce a peculiar fungus by cultivation of choleraic discharges in which bodies somewhat resembling "cysts " and " spores " existed, equally favourable conditions were at hand for the development of their ultimate elements ; — seeing, however, that the fungi which then appeared possessed no peculiarity, one may conclude that either the attempts to cause the development of the particular micrococcus of cholera were failures, or that no cholera " micrococcus" existed, at least not as the germs of a fungus. During the earlier part of the inquiry it was thought that a greater number of minute The statement that monads l)O(liosl )O( li os o f nil ovo-nrnV nntnvo p-snVorl and vibriones are more plenti- | JUUI^ vi aD. Ol^dlllC llcltLllC LXISLUI ful in choleraic than in other 111 cholera stools than Were f 01111(1 discharges nIiaAOTiiAVA • +n fliic immioccinn flio min^l elsewhere ; to this impression the mind was evidently, though unconsciously, predisposed, from the fact that the fermentation theories of cholera necessitated, to a more or less extent, the supposition that *monads, bacteria, and vibriones (Fig. lxiv) flourish to a * Irrespective of any theory as to the nature or mode of formation of these minute bodies, I have followed the example of Professor Hughes Bennett in adopting the terms " monads " when simple molecules are meant (Fig. lxiv — 1) ; " bacteria " when the bodies are slightly elongated (2) j "vibriones" when still more so; ami "leptothrix" when presenting a linked appearance (4). FIGS.LXIV LXIX DEVELOPMENTS IN ORGANIC SOLUTIONS greater extent in this than in other diseases. Thus far I have not found this to be the case; indeed, the discharges of cholera patients, if examined immediately, do not contain such quantities of these minute bodies, has not been supported by ospGC i a U y if the stools have been voided these observations. . -t •/ in rapid succession, consequently have not been loni? detained in the intestinal canal. Neither have I been able, after repeated observations, to find that, during the decomposition of a cholera discharge, a greater number of the minute bodies associated with putrefaction were developed in it than were developed The minute organisms as- under similar circumstances, such as Bociated with decomposition amoimt o f fo^ anc l mo isture, in of cholera stools not exces- . ' S ive ; ordinary alvme discharges ; nor have I been able to find that any peculiar growth, animal or vegetable, will proceed from the one which does not proceed from the other. On this point, however, the number of observations have been far nor microscopically peculiar. 00 f ew fllC SOIirCCS of fallacy being 2e^dt? ement " madG re " so many— to enable one to speak with confidence, but I trust in the next re- port to be able to furnish more minute data concerning this matter. On an average, out of a dozen experiments undertaken, not more than one is brought to a satisfactory conclusion, which is not to be wondered at, when it is considered that the quantity of matter experimented upon docs not exceed onefourth the size of a drop of water ; that this requires the free admission of atmospheric air, and that it has to be examined at least daily, for a month or more, Difficulties in ascertaining often for hours together. Either the nature of minute bodies ? n .-, -i-ii • ,1 are not only manipulative, the fluid Suddenly CVapOl'atcS, 01' tllO lens touches the covering glass, thus disturbing the geography of the preparation ; or, which is the most frequent accident of all, and one of the most untoward, a minute spore of some fungus falls from the air upon the moist slide — germinates ; the filament insinuates itself through the little air-orifice which had been made in the walls of the growing cells, and reaches the preparation, where it not only obscures the field, but alters the chemical and other forces taking place in that droplet, and the forms of life which had developed therein — I do not say spontaneously — become altered also. I have frequently observed that a slight disturbance affects the development of these minute 39 organisms ; either the forms of life previously present cease to grow, being replaced by others, or but dependent to a great 1C yjtal process becomes changed, extent on their susceptibility -in •f fisiYln lvnf nnflnno 1 /ImTrOrmnrl also set aside, but nothing developed in it. Illustration II: — About a drachm of ordinary fceecs was dissolved in am ounce of distilled water and filtered, Developments in more com- a portion of which Was placed ill a m\atter, ol vi^ n normai or a s ivi Ine1 ne watch-glass and boiled thoroughly ; a discharges. dror> of this was after wards placed in drop of this was afterwards placed in an ordinary animalcule growing slide, both being set aside under a bell-glass. On the second day monads and vibriones were present in great numbers in both prcpara- Young paramecia develop, tions, but on the third day they had ed on the third day in watch- „ 11 ¦?..,-.. ", • n glass, which had been pre- greatly diminished m number m the watch-glass, in which, however, during the night several young animalcule viously boiled ; belonging to the Kolpoda family (as figured at lxxx) had made their appearance. FIGS. LXXVI LXXXI. DEVELOPED IN SOLUTIONS OF ORGANIC MATTER LOOQ^ofan Inch, , « 300 In the live-box, however, vibriones only were present as before, some of them being very long, but no Kolpodse. On the fourth day great numbers of amoeboid bodies, varying considerably in size (Fig. lxxvi, 1), multiplying very rapidly, sometimes by leaving E amoeboid bodies ap- small fragments of their substance Lon the fourth day in I™!^/] /O\ . fhpnnvttnn PSPTnrn P 1 in me solution placed in a DCnmCL {*) , Tlie pOIUOn escaping m- clean animalcules cage. variably from a part near the con- tractile vesicle, which vesicle re- mained bright for fifteen seconds, became puffed out suddenly, as though it had been a taper, and remained extinguished for the same period, then gradually shone again. The detached portion (3) seemed not to be merely disgorged food, for it crept about the field like its parent ; it also divided into two, pretty symmetrical, halves. structure of the amoeba?, p or some time after the commcnceand their mode of multiply- , „-,... ,-, ? n ? mg. ment of division, the "nucleus is only seen in one half (4) after consider- able tugging, then coming together, then separating again, each time getting a little more detached, until in the course of about two minutes the separation is complete. Frequently a mass of granules is seen to intervene, probably indigestible particles, which may adhere to either half (5), but is soon cast off, and gradually a contractile vacuole is seen to appear in the second half, which creeping along the field draws particles into its substance, and acts in every way like its parent (6). On the fifth day the fluid in the slide having somewhat evaporated, a little distilled water was wat U e r r. OUS effect ° f addine added > when suddenly the hitherto more or less oval amoebae (Fig. lxxvii, 1) commenced protruding and retracting exceedingly long processes (2), which action lasted three quarters of an hour. They then became circular and still, except that the vacuole contracted (3). In another half -hour some of them commenced to creep along the field, disgorge themselves, leaving a string of granules to mark their path (Fig. *Z^^£^£T 1) ; others were observed in the course of another half-hour to become circular, with a clear halo-like ring surrounding them (2), their contents being in very active motion, reminding one exceedingly of corn in a miller's hopper. This lasted for twenty minutes, when suddenly all movements ceased; the halo and 45 vacuole disappeared, its outline became Method of disappearance of irregular an d undefined (2, 3) ; finally, the amoebae. ?,° , ? ' , , i although the eye was constantly observing it, all trace disappeared, and no distinction could be observed between other molecules in the field. The remaining amoebae seem to have undergone the same change, for when the eye was removed from the particular one described, none could be found, except a few empty-looking ones. I have frequently observed exactly similar phenomena occur in the so-called salivary corpuscles. Similar phenomena ob- ]$ 0 further change OCCUITed in the served in salivary corpus- ft former condition during the succeed- ing week. In the watch-glass the animalculse continued to increase and multiply, but other kinds did not Watc a n c . g O i f asl! esOlUtioninthe appear. The glass was held over a spirit-lamp and the liquid boiled, in order to see if out of their dead bodies others of the same or of another kind would appear ; but none did, and at the end of a fortnight the experiment was brought to a close. Illustration 111 : — The ordinary stool, to which allusion was made at page 33 as containing such quantities of Cultivation of the ordinary tlie an i ma l C uke in the " still" and stool which contained the , . ?. , . _ z . active condition, was kept under observation for six weeks. animalculse. A slide was taken and two minute portions were placed side by side, a distance of Two preparations on one a | JOut half ftn {nch ii n t erve ning, and circular covering glasses applied of the same diameter. During the first, second, and third days the changes which occurred were alike in the two preparations. The oil globules gradually disappeared, the circular, " still" condition of the animalculse became at first granular, ceased presenting the amoeboid projections, the latter being frequently not retracted, but trailed underwent similar changes i ji ni -iji i un tii along as they rolled under the glass ; the general appearance of the altered slide being represented at Figure lxiii, the earlier condition having already been described, and is figured at lxii. 46 tThe movements of the active little entozoon became more d more sluggish ; at the same time it became granular d circular, and finally disappeared altogether, probablypassing into the "still" condition, which also graduallydisappeared. The two preparations now assumed different appearances. (a). On the fifth day some fungi were seen to develop in one of the preparations, which may be designated — a; long filaments of the fifth day. oidium lactis, as figured at xxiii, commenced spreading over the entire preparation, and in the ap F pea^d n ino°ne? diUmlaCtiS midst of the molecules (which had also undergone various stages, as already described in the first illustration) little " heaps" were forming of precisely the same microscopical characters as are given at page 40 and other places. together with the little On the sixth day a few molecules in " heaps," in the midst of 1 1 • i , n n i 1 i • which y%ast ceiis appeared. the midst of the heap had increas- Ed in size, and on the eighth day nearly every heap was covered with yeast cells, in conjunction with very minute anguillulse (?) (Pig. lxxix). The oidium lactis disappeared entirely in the course of a few days, but no other changes took place for a month, except that No further changes. the yeast cells degenerated also. (bj. The portion under the other covering glass showed no evidence of fungal de- x2:^Z^r^ con ' velopment, nor yet yeast cells or anguillulce. On the sixth day ac- cumulations of perfectly motionless molecules had formed, especially near the edge of the glass, each heap possessing, as usual, a kind of central kernel with a more or less protoplasmic appearance; the molecules forming the peripheral part of the heap being quite as c^rTTsi^ HT iOn °°~ active as the molecules elsewhere. On the seventh day these heaps were crowded with cells of all sizes. Some of the molecules were larger than formerly; the greater number of the cells, however, were from about the size of a aS P rare C d\ C c° xx r t P Say? larbodies red blood-corpusclo to four times that size; the contents of the larger ones being more distinctly molecular than that of the smaller, otherwise no difference could be established between them. It is, however, particularly to be noted that the steps from 47 48 YOUNG PAHAMECIA DEVELOPED the minute molecules to the smaller sized corpuscles were by no means so gradual; it did not appear as if a sufficient number of molecules of the interven- a gap seems to exist be- ing grades existed t"o enable one to tween the diamater of mole- fhnf +Yip Invcrp nnvrvii /w+fm + i'tulipiWl hxr fhf» a white margin by the fifth nma > l 0 UIG GXtent indicated Dy 1 110 dark outline of the preparation in the day figure of the Maddox slide at lxxv. 51 52 DEVELOPMENTS IN SOLUTIONS OF In the midst of these molecules little heaps were seen to form, in which no motion was evident, nor yet any definite structure, but amorphous granules, around which, and above and below, myriads of monads and short vibriones played. Some slight distance from mSSr ° f tMs wMte lie margin an opaque line, consist- ing of unusually active monads, was seen separating the creamy ring into an inner and outer portion, but no distinction could be observed between the appearance of the molecules of one side from that of the other, except that in the outer the heaps were more plentiful, The monads, &c, in the central clear space had become perfectly still, and no heaps had formed amongst them ; towards evening the line seemed to have spread on either side, as the whole creamy ring became as thickly studded with molecules — consequently opaque — as the narrow line was in the morning. The circular cells seem to have disappeared altogether. On the sixth day not a single molecule quivered. The crcamincss of the margin had slightly diminished, but the "heaps" were still present, rather more slimy-looking, not so regularly circular, but frequently Some molecules near the i j. i i x. i • mi l • elongated and straggling. The dia- " heaps" enlarged, meter of some of the molecules in contact with the mass had. increased. A mycelial filament was now seen to insinuate itself from without into the preparation, having crept up the " chimney" of the growing slide, and but a fungus crept in and j_i n n ¦> -, , . -, , spoilt the preparation. the iurthcr development m or about the heaps came to a stand-still, although watched for a month. The mycelium spread in every direction, and gave rise to yeast cells. (bj. A similar slide containing a drop of this stool, to which a little of the growing solution JSSETrtS&S ££ of grape-sugar and phosphates was developed penieiiiium. added, went through the same stages as the foregoing, and developed into penicilliwn, as in Figure xix, 2. (c). A portion of the same preparation without a cover. . ? ing glass was preserved in a moist A similar preparation with- i 1 r\ Ti n • i i i• , out a covering glass chamber. On the third day a white speck was seen in the surface consist- ing of innumerable "yeast" cells (Pig. lxxxvi), with some filaments branching in all directions. On the fourth day ORGANIC MATTEIt: — BLOOD, SERTJM, &C. 53 tufts of penicillium had developed — two varieties (Fig. lxxxviii) — P. Glaucum (1), and P. Viride (2). This continued until the ninth day, when a few of the filaments springing up in the midst of the penicillium were tipped with a dewdroplike dilatation excessively delicate — a mere distended pellicle. In some cases they seemed to be derived from the same filament as others bearing the ordinary branching spores of penicillium, but of this I could not produced penicillium asso- \) G positive. TlllS kind of fructlficaciated with, a mucor-like form ? x . -i . -,-. , ? of fructification. tion increased rapidly, and on the fourteenth day spores had undoubtedly developed within the pellicle (Pig. lxxxvii), just as had been observed in a previous cultivation (page 13), precisely similar revolving movements being also manifested. The reaction of the liquid portion in the cell was Acid reaction of preparation. 1 • i_xi • j i l ¦• slightly acid, and became very much more so in the course of a month. No further change took place, except that the capsules became rather thicker, but never so resistent as to withstand the action of a drop of water, spores being instantaneously set free by it. In not a few cases a chain of spores, or sometimes delicate filaments, seemed to escape from these cysts, as if the spores within had germinated ; which indeed must have been the case, unless they had fallen from a tuft of e£S£X B S£r° nOf penicillium and adhered tp the capsule. In other cases dilatations (macro- conidia) appeared in the filaments, and even from these a chain of spores was occasionally seen (Fig. lxxxix).* fd.J A small portion of the evacuation was placed on an ordinary slide with a covering glass. It went through the same process as was described in connection with Maddox's slide (a), and eventually yeast cells were produced as at Figure lxxxvi, but nothing further. (c .) A similar slide placed in the same moist chamber presented similar changes as the fore- A preparation like the fore. going for the first four days. It Was going placed apparently j. i ±1 nn±_-i i j i under the same conditions not examined on the fifth, but when placed under the miscroscope on the sixth day, representatives of the Jwlpoda family, both active * In connection with the appearance of this mucor-like fructification in such intimate connection with penicillium on this and on other occasions, although merely an approach to the " cholera fungus" of Hallier — a fructification resemiJBEZJgZS? ™™z » ™ eh , more , cl T, Iy ' if . not id r tical with {t > haTin « been obtained under like circumstances from ordinary excreta — it must be allowed that it speaks very strongly in favour of the view so firmly advocated by this niycologist of a generic connection between penicillium and mucor. 54 SUMMARY AND and encysted, had made their appoarprodueed young paramecia. . " , ¦• ¦, j.i_ • ance in great abundance ; the various stages in their subsequent development corresponding precisely with what lias already been described in connection with experiments on ordinary excreta. Serous fluid, blood, and urine, from persons affected with cholera, as well as from other Experiments conducted persons } iaye |3 een i n H^q manner with, other substances. **?¦, , ? t subjected to systematic and con- tinuous observations, the air in some of the experiments having been made to pass through a red-hot tube before its entrance into the chamber in in some cases the admitted wni ch substances under examination air had first to pass through -i-it t i lj-iiT) a, heated tube. had been placed, as adopted by .Pro- fessor Tyndall, in order to destroy the minute atonies of organic matter which, according to this gentleman's researches, will pass through sulphuric acid or caustic potash undestroyed. The particulars of these observations are reserved for the present, the results being such that no benefit could be at- statement of results post- joined by giving them in detail. It is nevertheless hoped that the foregoing illustrations will sufficiently explain the methods adopted in investigating the subject of this section. The description of the changes which occurred during the cultivations has been condensed as much as possible ; more so than would be allowable, were they intended to establish any particular fact. A not unimportant lesson is, however, convoyed by even the comparatively few experi- JttSfJmSZSS. ments wllich J . IOTe been conducted, namely, that, in spite of more than ordinary care, very different forms of life will make their appearance in substances which are derived from the same source under conditions which seem to be identical, and that too in very simple mixtures. Consequently, the greatest caution must be exercised in estimating the importance or otherwise of any peculiar manifestations of vitality which may be observed in substances associated with disease. The results of the investigations Summary of conclusions drawn from aii the observa- referred to in this report may be tions. ii „ „„ __«_ J »J . thus summarised : — 1. That no "cysts" exist in choleraic discharges which are not found under other "Cysts." conditions ; 55 CONCLUSION OF PART T. 2. That cysts or -sporangia" of fungi arc but very rarely found under any circumstances in alvine discharges ; Sporangia. 3. That no special fungus has heen developed in cholera stools, the fungus dcs- Special fungus. opeuwi mngus. Mil tt it i j_ • 1 enbed by lialhcr being certainly not confined to such stools ; 4. That the still and active conditions of the observed animalculoo are not peculiar to this Animalculse. disease, but may be developed in nitrogenous material even outside the body ; 5. That the flakes and corpuscles in rice-water stools do not consist of epithelium, nor of its debris, but that their formation appears to depend upon the effusion of bloodplasma; and that the "peculiar bodies" of Parkcs found therewith correspond very closely The noecuient deposit, and in their microscopic and chemical Parkes' " peculiar corpus- r .l invnf ,j- OV a n i 1 1 with worst period in the and in Lower Bengal generally, is, according to Dr. Pettenkofer. due to the fact that in the former Upper Provinces. place the ground for the greater part of the year is too dry, there being no rain, and the water being commonly many feet from the surface ; whereas in Lower Bengal when the rains set in cholera ceases — the ground becomes too wet. Cholera is worst in the latter when the water-level is at its lowest, namely, about April ; whereas in the former cholera is at its worst when the water-level is rising or about subsiding (August and September), so that, I presume, the Munich Professor would explain the reason why cholera is endemic in Bengal, and only epidemic in the Upper Provinces by the fact that the wet season is much shorter in its duration than During the rains it is too the dry, consequently the conditions wet for the development of necessary for the development of the the poison in Bengal, and at . " i i • i j • i other times it is too dry in pOISOII OCCUr Ollly during sllOl't periods, the upper Provinces. whereas the same conditions affect Lower Bengal in a different way, giving rise to short periods of exemption, instead of the short periods of attack, consequently the inhabitants of the "Bitch" are more exempt from cholera when it overflows with water. It must also be borne in mind that a local fall of rain is not in all cases the only cause of variation in the height of subsoil water, for an adjoining river may rise or fall, irrespective of local conditions, and perhaps give rise to an alteration in the amount of moisture present in the soil. In a great number of instances, however, the level of the water in a well adjoining a river is considerably above that of the river, as a non-porous, clayey layer may dip towards it, thus confining the water to its bed. Local rainfall not always a Again, lieaVV falls of mill Oil distant true criterion of the amount ,• V , p,, " , ? , , „ n . of soil moisture. hills may afreet the level of the water in the plains, should an impermeable stratum extend from tlie one to the other, over which water might flow. These and many other such facts connected with the geology and the topography of a place must be carefully considered before any opinion can be formed of the 59 THE POISON IS DEVELOPED IN THE SOIL. correctness of the views advanced by this distinguished Professor. It will be seen from the foregoing that the poison is considered not to develop in water, The choiera-germ does not which is contrary to the commonly develop in ¦water, nor in the ¦% • • -i •, ?¦ , human body, received opinion, nor docs it multiply to an appreciable extent in the intes- tinal canal, the human body being merely the stage upon which this actor plays its part. The poison requires a special nidus in which to multiply and to develop into infecting matter. This Pettcnkofcr traces to the soil, especially to alluvial soil, but in the soil. which, being so exceedingly porous, allows free interchange between the air in its interstices and the air above, as well as being subject to a great variation in the amount of water which it contains. Whilst marking out on a map the places suffering from cholera, he was particularly struck Tendency of cholera to at- w ith the predisposition it seemed to tack places situated along .„ . \, nl,,n 1 ,, . ? . , rivers. manliest for following the natural water-courses of the country, rather than the usual routes of traffic. In the former, the places of attack were pretty regularly situated, whilst along the roads for intercommunication, the affected places show great irregularity, cholera spreading only in those parts in which the soil was of an alluvial nature, although quite as many opportunities existed for the dissemination of the poison by means of intercourse in the places never attacked as in the less fortunate localities. The cholera-germ, as described by Pettcnkofer, may be defined as a specific leaven, requiring The choiera-germ supposed earth, consisting of organic matter to be ol' the nature of a fer- , ? ... ° ?° in ment> and salts, with a certain amount of w r ater for its development to infec- tious matter, just as other ferments require certain special substrata and moisture before it manifests its action. If ordinary leaven be added to sand no action takes place ; if it be added to dry flour, it does not spread beyond the immediate vicinity in which it was placed; but if the flour be moistened " the little leaven leaveneth the whole." The question naturally occurs — By what means does it get into the human body after being uSedin°twowrvi?- Semi - thus developed in the earth ? To this Pettcnkofer replies : There are two 60 HOW THE POISON IS CONVEYED FROM THE GROUND. ways by which substances may arise from the ground, even from a great depth :—: — means of (1) the water, and water and air. by means of (2) the air contained in its interstices. Numerous illustrations may be produced of the possi- bility of substances, perceptible to Possibility of the air be- ie olfactory nerve, making their way coming contaminated by sub- ii? • i in ixi stances rising from beiow. upwards from considerable depths, such as when a sewer bursts, or an escape occurs in a gas-pipe. Frequently this fact is not observed where the mischief has taken place, but in a house, perhaps, some distance from it, the warmth of which, should it not stand on an impervious layer, attracts the disengaged matter like a chimney, and the house acts as an escape-pipe for a noxious gas. Were it ever conclusively shown that cholera depended upon some fermenting process taking place in the ground, which had been originated by some of the poisonous material being placed in a soil adapted for its development, it would follow that in India large substantial buildings would be safest by day, being cooler than it is without; but, unless the Tendency of foul air to flooring were made impervious to enter heated rooms with per- . . . ° i-ii ,1 , n -i -, meabie floors. air, it would be the most unfavourable at night, being warmer, consequently the native's hut approximating more closely to the temperature of the air ought to be more exempt from cholera. Although cholera is not considered to acquire its pro- perty of infection by being developed Water, a vehicle for the • W ofp V still wntov n«s wpII i« hit dissemination of cholera. m ™ atei > Sim V\ aiCl as Weil as air may act as a vehicle conveying the infectious matter from the ground, consequently this theory in no way affects the importance to be attached to the value of obtaining water from a pure source; indeed it speaks very strongly in favour of obtaining it from places as far removed as possible from human habitations. Particular attention has been drawn to this subject very lately* by Dr. Buchanan, one of Buchanan's explanation of i.T,« op V prnl rlkHno-nkhprl Snnitnvv Pettenkofer's theory. lllL s 6Veiai dlStingUlSHCd Oanitaiy Officers whom Mr. Simon has gather- Ed around him at the Public Health Department of the Privy Council. While allowing that there is a connection between the disease and the level of the water in the wells, * Medical Times and Gazette, 1870. BUCHANAN ON PETTENKOFER's THEORY. 61 Dr. Buchanan maintains that the mischief is in the well itself; because "it is precisely at the period when soil water is sinking that wells sunk in porous soil must, if ever, furnish impure supplies. A well in porous soil gets its water in two ways ; water stands in it up to the level of the soil, and also drains into it from every source (from rain, from slops, from cess-pools) on a higher level than that of the water of the soil for many yards around. In other words, besides receiving water from the The deeper the well, the general Watci'floW through the SOll, greater the area of surface -j. voflf >j vps t ] lo ] aO n] c O il woW «sOilcpollution which is likely to 1U IW'LIVLS HIO lOCdl bOII AWllLl, SOdK- ing from a cone of ground of which the surface of water in the well is get into it. the apex. Let the level of water in the soil be high, and the base of this cone is small ; let the level of the soil water be low, and the base of this cone (at the surface of the ground) is large. In either case the saturated soil is comparatively impervious to more water, and approaches the condition of a non-porous stratum. When the soil water is at its highest therefore, impure slops and excrement that may be on or in the ground tend to run horizontally away. When the soil water on the contrary is low, such matters tend to soak downwards." It will be observed that Dr. Buchanan testifies to the matter-of-fact portion of Pcttcnkofer's statement, namely, the connection of certain diseases with the level of the soil water, but explains this connection in a different way. Buchanan produces very remarkable illustrations in proof of his statements, which will certainly be borne in mind whilst investigating this subject in India. The possibility of the foregoing being the true explanation of the connection between cholera and the level of the soil water had not wens'aTiow. °* ***" when escaped Pettcnkofor, as he states that examinations have been made of the quality of water which is sinking, and the results were by no means unfavourable — in some cases the water was even found to be more pure. As far as the tanks in Calcutta are concerned, I cannot bear testimony to the observations of Pettenkofer in this matter, because the percentage of organic matter has been greater when the tanks were low than when in the contrary condition ; concerning the quality of water in deep wells at various heights, I possess no data. The foregoing remarks will, I trust, be found to present a tolerably clear exposition of the theory concerning the relation 62 ITS RELATION TO THE lUVETIS. said to exist between the spread of cholera and the state of the ground water. I now proceed, in as few words as possible, to give an account of my visit to the places affected with cholera in the North-Western Provinces during the severe epidemic of last autumn. ALLAHABAD. I arrived at Allahabad towards the end of August, in accordance with the instructions I had received, in order to accompany Arrival at Allahabad. the Sanitary Commissioner with the Government of India in his tour through the cholera affected districts, and thus be able to avail myself of his advice and direction. Seeing that our visits to the various places, to be hereafter alluded to, were necessarily of short duration, it was impossible for me to obtain more than a very superficial knowledge of the geography of a place extending over such a wide area as Allahabad docs. Situated in the angle formed by the junction of two rivers, the Ganges and Jumna, it Relation of Aiiahabad to wag thougllt not i mpro bable that the high or low condition of these rivers might materially affect the level of water in the wells, seeing that many acres of land are swamped during the rains, the station being almost surrounded by water, as a glance at the accompanying map will show. This, however, was ascertained by Dr. Bow not to 1/ — — be the case, at least as far as the The level of the subsoil Jumna was concerned, the water in water nearly 30 feet below ? 11 i • ir»rvnj-ii that of the jumna. the wells being nearly 30 ieet below the level of the surface of the Jumna. The average depth of the wells from the surface, as examined by Dr. Chalmers and myself, was found to be from 50 to (35 feet. The average variation in the level of the water between the dry and the wet season is about ? , 10 feet, whereas the Jumna varies to Extent of variation of level. ? iionn.Pi t the extent of 30 feet or more under ordinary circumstances ; nor does the alteration in the water-level of the one correspond with the variation in the other, and a consecutive fall of rain of 12 inches will not raise the level of the water to the extent of more than one or two; a great portion, doubtless, finding its way into the river before getting into wells, especially after the first > falls of ram, when the ground does not permit of such free percolation. It cannot, therefore, be said that the amount of subsoil water at Allahabad is materially affected by the rivers which bound it on either side ; t^LZT not affe ° ted by nor will it be safe to judge of the extent of moisture present in the subsoil by the registration of the well water alone; the rainfall must also be taken into account, as the latter at Allahabad appears to influence a ffe:S; n th O e f rSaif iefly the condition of the soil more than the permanent subsoil water. The soil here is of a sandy, clayey nature, intermixed with layers of kunkur. In the hot weather extensive fissures are to be observed everywhere in the ground extend- Porous nature of the soil. . •; , , ? ° , ? , ing to great depths, and exceedingly permeable to water. On subsequent examination, it was found, when dried in the sun, to be solid to the extent of one-half, the other half being interstices filled with air. In order to have a more precise knowledge of the extent of the porosity of the soil upon which the various barracks have been built, and which are said to vary in the degree of their liability to cholera, although in other respects apparently as like one another as it is possible for buildings to be, and the sojourners therein Cholera said to affect some enhippf \r\ thwhsipl v \\\o enmn infln barracks more than others. SUDjeCTJ 10 piCCISeiV UIG Same lllllU- ences as regards food, clothing, and water, it was thought that perhaps some clue could be obtained by ascertaining the extent to which the soil beneath the buildings was permeable to the air below. General Tr avers, v.c, immediately permitted samples of this soil to be obtained, which on my return to Calcutta were subjected to the following treatment : — A little of it was reduced to moderately fine powder in a mortar and placed in the sun until thoroughly dry. In the meanwhile, two burettes were fixed on to a stand, the lower portion or point of one being connected to that of the other by means of a piece of India-rubber tubing supplied with po^os?tfofsoirs! rtaininethe a C 11 P> S0 aS t0 b ° al)le at Will to interrupt the connection between the two tubes. A given quantity of soil (100 cubic centimeters) was carefully placed in one burette, and a similar amount of water in the other. The latter was allowed to How into the former, which, as it ascended in the tube 63 64 CHEMICAL EXAMINATION OF THE SOIL. containing the soil, was seen to drive out the air existing in the interstices, the amount of air displaced correspond ing to the amount of water which entered. When the watei came up to the upper edge of the soil in the tube, the connection between the tubes was interrupted, and the amount of water used read off. As a few of the particulars of these observations may be useful for comparison when more exact data shall have been obtained Result of experiments. of the relative liability of the barracks in question to cholera, I append them in a tabulated form : — Permanganate solution Soil at a depth of 4 feet from f. " /,;,'.,' solution of loz. soil, n iuu puti loozs. water requiring by measure. 4decema o f the sam? solution. f Clydesdale Lines, No. 8, south end . . . 50' 5 decems „ „ „ 8, north „ ... 46*4 4 do. „ „ „ 3, south „ ... 533 8 do. 'I „ „ „ 3, north „ ... 50' ..... 'rt •{ Chatham Lines, No. 8 ... ... 50' 5 decems ' Artillery Lines, No. 2 ... ... 50* 4 do. Wellington Lines ... ... 4G'4 6 do. New Cantonment Barrack, No. 3 ... 4G"4 4 do. I Jail... ... ... ... 533 5 do. Six of the specimens were subjected to chemical examination with the view of ascertaining th C e h s e oTi! Cal examination of whether the soil near the barracks, at or about three feet from the surface, contained an unusual amount of organic matter or not. One ounce of soil was taken and allowed to stand for twelve hours in pure water, shaking it a few times during this interval ; it was then filtered, and the clear solution examined in the manner usually adopted for the examination of water. The results were pretty much the same in all cases ; except in those where the soil had been a " made" one, 65 MICROSCOPICAL ¦ EXAMINATION OF THE SOIL. the amount of lime-salts varied, but I was surprised to find that the soluble organic matter, as estimated by a standard solution of permanganate of potash, Amount of organic matter r ]-,] ynnnh ovporvl flip nmmmf in the soil was not excessive. aKI n^.^lUCll CXCCLd me aniOUllt present in the ordinary drinking water of Calcutta when estimated by the same solution. (The exact relative amount of organic matter present may be ascertained by reference to the foregoing table). From these observations, therefore, I infer that in the ground beneath and about the barracks at Allahabad, both in the old and new cantonments, the amount of oxidisable matter was not in excess, at a comparatively short distance from i he surface, at the Inference. time when cholera visited that station; consequently the epidemic could not have been owing to putrefying matter in the soil of the cantonments, unless such matter had been washed into the wells by the rain, and thus infected nearly a hundred and fifty persons belonging to the European troops , stationed there. To have produced this, the amount of surface pollution present before the rains set in must, I should imagine, have been very extensive indeed. It was also thought desirable that a few samples of the earth should be taken and moist- Microscopic examination of cnC( | witll water J n ord(jr to ascer t a in whether any special form of life, animal or vegetable, would make its appearance. I select two examples. A small portion of dry-earth from the new cantonment was placed in a test tube, to which a little water was added, sufficient to cover it. During the first and second days no particular forms of life were observed, but on • the third and succeeding days several on the third day ordinary mmute infusoria had become revived, mlusona appeared. , . ' and presented exceedingly active movements (Fig. xc). A similar portion of soil from the Clydesdale Lines was treated in the same way. In it also no particular objects were manifest for the first two or three days, but towards the end of a week, in addition to the objects delineated in the last figure, bodies in the circular, still, and active condition — not in any way distinguishable from the animaleulsß in another sample the ant- already described as occurring in malculoe found in evacua- i -i • i j.i i ¦ i tions were plentiful. choleraic and other discharges — were seen to have developed in great 66 VISIT TO CAWNPORE. numbers, some freely moving in the fluid, and others imbedded in granular matter (Fig. xci). Nothing further was observed in any of the samples, nor could I detect any evidence of the existence of the ultimate elements of fungi. I also accompanied the Sanitary Commissioner to the "cholera-camps" occupied by the 58th Visited the cholera-camps, -n • , 1 , n /•! ' 1 n Regiment, about nity miles irom Allahabad, on the Jubbulpore road; and Dr. Chalmers, the Deputy Inspector General of Hospitals, very kindly undertook to show me nearly every part of the the native portion of the city -j_ i po^f™™™* T o t) r Twino* and surrounding villages. City dJIU CUlloniriClll. J.O Ul. ±lYing also I am indebted for similar help. These excursions were undertaken more with the intention of getting a fair insight into the geography of the place, than of ascertaining what the exact sanitary arrangements were, — to report upon which not being the object of my visit. Careful notes, however, have been taken of what was seen and heard concerning the outbreak of the epidemic, but their narration would unnecessarily prolong this report and answer no good purpose. I hope, however, on a future occasion to turn what I then learnt to account. CAWNPORE. On the way to Lucknow, a few days were spent at v, H ittocawnporo. Cawnporc. Compared to Allahabad, the troops m this station had suffered very little. Dr. Bryden states that the admissions were 27 and the deaths 17. The soil at Cawnporc is very General nature of the son, like what it is at Allahabad, hut contains less kunkur. Near the artillery barrack there was more clay than elsewhere, but, as a rule, the ground is very permeable to water. Many of the wells are very much nearer the surface, water being found at five or ten feet, in- Wolls, and tlioir relation to i- \ v{? ex. • ±_ jaui the river. stoacl of fifty or sixty as at Allaha- bad. Nevertheless, some of the wells examined were thirty feet below the surface. Such a variation I did not observe at Allahabad. The ground slopes towards the Ganges : I could not ascertain whether the rise or fall in the river affected the level of the water in the station ; but when the river rises, it swamps a largo portion of the country along Kb banks. FIGS. XC I. DEVELOPED IN MOISTENED SOIL FROM ALLAHABAD XCII 111. LUCKNOW XCIV FYZABAD XCV MEERUT The cholera-camp of the 14th Regiment was pitched at Bhowpore. The ground about this camp was more sandy even than at Cholera-camp. Cawnpore, and more permeable to water and air. The relative degrees of permeability of this soil and of the soil near the barracks occupied by artillery, cavalry, and infantry regiments are given below : — Permanganate solution Amount of air squired to give a per,w !. i , n nianent tint to a solu- Soil at a depth of 4 feet from mrts v . sure iUm of Iil > 10ozs 10ozsof sol water ntptaag 4de-01 bOU< cemßof tlio same solution. f Cholera-camp, Bhowporc ... ... ... ... 533 6 decoms g „ ltesideney ... ... ... ... 4 gnme solution, I „___ ' No. 1 Barrack, occupied by Royal Artillery ... 3 50 1 0 docoma ?2 „ „ „ „ ... 3 533 10 „ .. 2 „ „ „ „ ... 6 533 10 „ Hospital „ „ „ ... 3 533 8 „ ? ... 0 53-3 No. 12 Barrack, occupied by 62nd ?. ... 3 533 10 ? i „12 „ „ „ ... ... 6 60' M- „ 2 „ „ „ ... ... 3 80" 12 „ „ 2 „ „ by 102 nd ... ... 3 633 12 „ „ 1 m » », ... ... 3 50- 6 „ „ 0 „ „ „ ... ... 3 60" 14 „ ? 3, Married Quarters, sth Lancers ... 3 50 50-„ 2 „ „ „ ... ... 3 lfiO 0 „ Jail, No. 7 building ?, ... ... 3 50 1 5 n „ „ „ ... ... ... 0 50- Several specimens of soil were examined microscopically, but nothing could be detected Microscopical examination. . V . . , ° ? _, ? _, . m the moistened soil for the first two or three days, presenting unmistakable evidence of vitality. Infusorial animalcules of many kinds gradually appeared, but I could not state that any marked diiFcrenccs existed in the various specimens observed. A figure of those which revived in some soil, from No. 2 married quarters of the sth Lancers, will serve to illustrate what these were (Pig. xcii). In one sample, however, some very interesting low forms of life appeared, about which Mr. Huxley and Hackel have lately written so much. The test tube in which this particular sample (from a depth of six feet in No. 2 barrack, occupied by lloyal Artillery) was seen was fortunately a very thin one, and permitted the use of a high power when placed on the stage of the microscope. The bodies observed consisted of minute masses of translucent, colourless jelly, without nucleus or contractile vesicle; a low form of life, corres- i n short, not the slightest evidence S&SJX."""" d6S " of structure existed. Their move- ments were very slow, slower than ordinary amoebae, and being translucent, it was only by careful illumination that they could be watched. Two of them are sketched at Figure xciii, in the act of protruding long processes of their substance among some animalcule which have become encysted on the walls of the tube. This moving substance presents precisely the same microscopical appearances as the hyaline, glary matter surrounding the encysted bodies. The little colony depicted of the latter was watched for several weeks, but no changes took place, consequently the nature of the encysted bodies could not be made out. It is very remarkable that such bodies retain their vitality so long, as they must have imbedded in this dry soil for several years. FYZABAD. There were a few cases of cholera in this station also, but fortunately only two deaths oc- site of cantonment at curre( j amon g the European soldiers. Fyzabad. __ . ° , . ? , _? ?, The cantonment is situated on slightly elevated ground on the banks of the Gogra, but no part of it is swamped by this river, nor Relation of the river to the j t believed tlmt the rise and f all of the river affects the condition of the wells. The soil is sandy everywhere, except near the bed of the river, where there are more traces of clay. Here and there a layer of kunkur is interposed between the upper more clayey layer and the lower one. A few samples of the soil were preserved for subsequent examination, the result of Examination of the soil. i • i i . ? , ¦. -, which may be seen in the table below. On being microscopically examined, nothing which could possibly be construed as having the most remote connection with cholera could be seen. No spores of fungi could be identified, and the infusoria which became revived in the course of a few days were of the ordinary kind (Fig. xciv). 70 71 Permanganate solution re- Amonnt of air quired to give a pcrmn„.., , ? „. „ , r contained in neiit lint to loz. soil, Soil at a depth of 4 feet from 100 parts by lOozs. water requiring measure. 4 deeems of same solution. ¦ C No. 13 Barrack (11th Regiment) ... 5O 5 deeems Fyzabad.-j No. 17 Barrack (11th Regiment) ... 60" L Hospital (11th Regiment) ... 533 AGRA. This station escaped with one death from cholera among the European troops, but the EropZs^AgraTMsVea" natl ™ population in tllO ]afl U Well as in the bazaars suffered considerably. Dr. Christison very kindly showed me over the whole station, so that, in spite of the short- Filthy state of ravines. ' ? ... x ™ . . , , ness of the visit, a iair idea was obtained of its physical geography. As at Lucknow, ravines intersect it in every direction, and for the most part contained filth. In connection with this subject, there is a popular belief amongst the more intelligent native community that, when the river Jumna flows on the city side of a sand embankment which has formed in its bed, cholera does not prevail at Agra, as the river popular belief concerning carries all the filth away, but w r hen the influence of a sand-bank •, fl nwq on f} lp o fV Sl - f ] o J-U p f li<,p nqp i H in the bed of the Jumna op- ir nows on lLlt oir hlu^> tile UISCdSC IS posite the fort. more liable to make its appearance. There may or there may not be something in this ; there is, however, a serious objection to the river flowing on the city side, on account of its tendency to undermine the fortress. The wells are very deep, fifty to sixty feet, and the water brackish, but whether the Wate P r! h * weUs ' brackiSh depth of the wells is governed by the amount of water in the river, I was unable to ascertain. This information, however, will shortly be obtained in connection with the registration of the waterlevel established here as elsewhere. MORAR. When this station and the adjoining fortress of Gwalior were visited, nearly a hundred deaths from cholera had occurred among the European troops, aw a °Sort at M ° rar and although it was formerly considered one of the places exempted from epidemics of cholera. They had suffered severely in April, and still more so in August, sunstroke having been exceedingly prevalent among the native population as well as cholera. Every effort was made to get all the information possible concerning the epidemic, more especially relating to those points which seemed to bear upon the question as to whether or not the origin and spread of the disease had any connection with the ground upon which the people stood. The cantonment is situnte on a low-lying plain, sur- rounded by numerous hills on three to^iveli 011 ° f cantonment sides, with the river Morar on the other. Some of the barracks are situ- ated below the level of the river, so that the drains have to be taken in another direction. Other barracks, such as the ones allotted to the Artillery, are about seventeen feet above the level of the Morar. An embankment has been erected across the bed of the river, so as to provide the station with a sheet of "orna- mental water/ about a quarter of a mile at its widest part, and increas- The artificial lake ing the depth of the river for about two miles above the dam. It is not improbable that Dr. Pcttcnkofer, had he been here, would have made minute inquiries as to the extent of moisture supplied to the neighbouring subsoil by this artificial lake. The wells arc from twenty to forty feet deep; the variation is said to be about five. JJopth and variation of The water ig consiclored to be good. Dr. Whitwell has examined it very lately, and has kindly favoured me with the particulars of the analysis of an average sample ; in this there is not a large amount of Quality of water. organic matter, and no excess of deleterious salts. There arc two kinds of soil at Morar, the red and black soil ; both contain pcrsalts of iron, with lime and magnesia, but no nitrates nor nitrites, as one woyjd have expected to find, had the ground Black and red soil been tainted to any great extent by the ordure of other days ; nor was the amount of oxidisable matter, as ascertained by the permanganate of potash solution, by any means 72 THE ROCK AT GWALIOTI. 73 ¦ssive, indicating that the barracks and their surroundhad not recently been subjected to contamination. ¦ The " black soil" was not universally distributed over the surface. Many yards of excavanot regularly distributed ; ? .*,«'. , . , , tions were examined m which not a trace of this kind of soil existed ; in others, again, a stratum of it was seen extending for long distances ; at one end the layer might be ten feet, or more, in thickness, gradually diminishing until it was finally lost in the red ; below those, a gritty sandy layer exists in which water is found. The foundation of several blocks of buildings, which were about being erected, were seen to present this uneven distribution of black and red soil, consequently the floorings of such buildings will vary hence a barrack may stand • ru nvfpnf tn wlllVll tIIPV IVP on soils of varying porosity. in UIC OXlOnt 10 WHICH UlCy aiC permeable to gases, &c, from below ; because the porosity of the red earth is considerably greater than that of the black. If Pettenkofer's theory be true, a building placed on this black clayey soil ought to be in a better sanitary condition than those built on the red — other things being equal. The relative porosity and amount of organic matter may be ascertained by reference to the table at the end of this paragraph. The samples enumerated arc only a few of the ones examined, General Vaughan having most kindly procured specimens from every portion of the cantonment. The cholera-camp was four or five miles out of the station, near the summit of two or Cholera-camps. three little rocky hills, the hospital apparently having a little hill for itself. The For lr ess of Gwalior is about six miles to the west of the cantonment of Morar. It stands on a rock whose summit is about If mile in length and about \ mile across in its widest part, and Fortress of Gwalior. from 300 to -100 feet high, the ascent to which is very steep. Immense fissures may be observed in the rock whilst ascending the steep towards the gate at the entrance of the fort, these being for the most part filled with earth. On entering the fortress, nothing is seen but huge blocks of buildings standing on a barren rock strewn with a few half-withered trees, or rather shrubs. The surface of the rock is naturally very uneven, stone forming the found- A barren- fissured rock ? * „ i p 1 *i i* i-ij ation of one end ol a building, whilst frequently " made" soil, to the depth of twenty or thirty feet, K. 74 VISIT TO MEEIIUT. forms the foundation of the other. The rock itself is a sandstone; splits to any extent, and very easily worked when wet, but excessively hard when dry. It is porous to the extent of one-third of its bulk, consequently able to retain a great amount of any sewage that may be thrown upon it. of sandstone. The heat on this rock is very great ; it is much com- plained of, especially as it continues Temperature. during nearly the whole night, be- cause by the time that it begins to cool, the rays of the sun arc directed towards it again. The samples of soil obtained consisted entirely of rub- bish ; there does not seem to be an Wo original soil. j\o original son. n • ,1 i t i. i i i inch in the place undisturbed by man until the bare rock is attained. Table showing relative porosity and organic matter in the soil. Permanganate solution re- Amount of quired to give air in 100 a permanent Boil iron) Depth, parts of tint to loz. soil, soil by lOozs. water measure requiring 4 docoins of same solution. ( (lied.) No. 4 Barrack, occupied by B. A. ... ... ... ... 3 16 # O 8 deccms Ditto ditto ... ... ... ... (i 406 9 „ No. 9 Barrack, occupied by 103 rd ... ... ... ... 3 66 1 6 „ >^ No. 2 iJarrack, ditto ... ... ... ... 3 ... 5 „ | •{ No. -1 liarrack, occupied by R. A. (Yellow clay) ... ... 3 46' 4 „ <( (Black.) No. 2 Barrack, occupied by B. A. ... ... ... ... 3 31- 5 „ JMtto ditto ... ... ... ... 6 86 1 4 „ L No. 6 Barrack, occupied by Md. Qrs., B. A. ... ... ... 33 (i „ MEERUT. Cholera visited this station in September and the beginning of October, having been pre- P^codeXTafn. at Meemt ceded by a heavy fall of rain. Nine- teen cases occurred, with fourteen deaths, among the European soldiers, whilst about a hundred cases occurred among the natives of the bazaar. The cantonment is situated on a large plain, with scarcely any fall, consequently not admitting of good natural drainage. There is a deep ravine separating the European and the Native lines, on either side of which for a short distance good clay is found ; otherwise it is rather Nature of soil. sandy everywhere, quicksand being frequently met with in digging the foundation of a building. FIG XCVI DEVELOPED IN MOISTENED SOIL FROM MEERUT FIGS. XCVII Cl. .. ? PESHAWUR attained at about 10 to 12 feet from the surface, the extreme variation in which is, according to Dr. Berkeley, about five feet. Rain rapidly affects the level of the water in the wells, the amount of rise of the latter being almost equal to the fall of the former. This is the reverse of what occurs at Allahabad, where a great portion of the rain- fall either drains to the river or is intimate connection be- evaporated before reaching the pertween surface drainage and * . , -, , m , . P . .. x , the weiis. mancnt water-level. This intimate connection between the wells and the surface at Meerut is of great sanitary importance. Seeing the ease with which any sewage may get into the wells, and as the condition of the ground does not permit of free natural drainage, it is self-evident that the greatest attention should be paid to remedying this defect by artificial means. In the more minute examination of this soil, subsequently undertaken, no evidence existed of the ground in the vicinity of the barracks being in Relative amount of organic a polluted COllditioil, and On tllC wliolo matter and porosity of the x ? , ? ? .. S oii. was rather less porous than the soils already alluded to, with the exception of the black soil at Morar. Permanganate solution Amount of air "''l"' l^' Jotf™ " nnntnlnniUn permanent tlllt to a Soil at a depth of t feet from 100 ,',' • s V, wlutlon of loz. soil ; measure 10 ozs - water re 'l - meaßnre. ta«f4deoemß of lame solution. f Between Nos. 44 & 46 (105 th Regiment). 45" 5 decems „ 34 &39 „ „ ... 50' 6 „ 13 & IS „ „ ... 50' 5 Meerut...-j „ „ 1& 2 (4th Hussars) ... 55 - 5 „ „ 13&14 „ .. 46-3 5 Married Quarters, No. 16, R. A 50- G „ This soil was examined microscopically in the same manner as the others were, with somewhat similar results. During the first few days its solution contained no infusoria, at least not in motion, but subsequently they made their appearance in great numbers. These in one sample, namely, 78 76 PESnAWUR SOIL. in the soil from between Nos. 1 and 2 blocks, occupied by the 4th Hussars, consisted almost Microscopical examination entiroly of various p l iases in t \ 1G exist _ encc of monas lens (Figs, xcv & xcvi). These alter their form very rapidly, frequently protruding an amoeba-like vesicle, as seen at Nos. 1 to 5, Figure xcv, which represents one animalcule assuming different forms. There are also great numbers of very minute amoebae (0) which seem to be an earlier stage of this animalcule, and when it gets older it becomes elongated (7-8), sometimes acquiring two filaments. They are frequently seen to multiply by division, as seen in Figure xcvi, where No. 3 runs through the stages delineated at 4 to 7 in the course of ilvc minutes, the two at 7 becoming as perfect in all points as the original one. The green bodies in the figures, which rolled about the field, are algcc. PESIIAWUR. The Sanitary Commissioner having subsequently visited Peshawur (where over 350 cases of cholera were reported as having oc- Soil from Peshawur. curred during the month of September among the European troops alone), favoured me with two samples of soil, one sample from a depth of three feet, and the other from a depth of six. It was in hard lumps, of low specific gravity, owing to its spongy nature, exceedingly like a piece of pumice stone, and when Spongy nature of soil. applied to the lips, so freely could air be made to pass through, that a feather placed on one end of a table could be readily blown to the other. Its solution was slightly alkaline, and contained rather more organic matter than the average, Its relative porosity and 1 n j-i l • • l as may be seen from the subjoined organic contents. table. Permanganate) ' solution required (o Soil from a dunth of Amount oi air in li'O parl s ffivo a permanent tint to a solution ol 1 by measure loz. soil, looses, water requiring t doccms of same solution, ?( X oi>- in doceras "I ( i FIGS. Cll CVI DEVELOPED in MOISTENED SOIL FROM PESHAWAR VARIOUS FORMS ASSUMED BY ONE IN TWO MINUTES FIG. CVII FOREGOING BECOME SPHERICAL & STILL It was subjected to a prolonged microscopic examination. During the first three days a number Microscopic examination. „ H -, -, -, , " . ? n'i of molecules developed in the fluid containing the three feet soil ; then an abundance of animalcules like the ones alluded to in connection with the Mccrut soil (Fig. xcvii, 2-3). At Nos. 5, 6, and 7 various forms are depicted, assumed by one in two minutes, which was also occasionally seen to jerk suddenly in the same manner as 2 and 3. The reddish body at lis a spore, probably belonging to the Dematiei family — a very common fungus. The test tube containing the other sample of soil from a depth of six feet having been left Another of Hiickel's Moners (?) -,. x . . , „ , ° undisturbed lor a week, was, on examination, found to contain several examples of slimy bodies of a lower organization than the amoeba, there being no contractile vesicle, although generally one or more vacuoles were seen (c). Nearly all of them contain molecular matter, which flows towards the portion of substance in the act of being projected. Figures cii to cvi illustrate the various forms assumed by one of these in the course of twenty minutes. They were not seen to divide, nor did the protruded processes become amalgamated when they crossed each other. A great number of vibrion.es developed in this solution, more so than I had observed v™° neS unusually pre " in any of the other specimens of soil examined, and were vcrv active. These arc figured at ci, amongst which one of the just described moners is seen with extended processes, which were observed to wander throughout the fluid something like the " horns" of a snail. To these processes monads and small vibriones adhered, which were drawn into the substance of the moner as the processes were retracted. Monera become spherical. mi , Jr , ? . Three days afterwards, all the moners had become spherical and perfectly still (cvii). The other animalcules which made their appearance were those commonly met with, and require no special description. They are figured at xcviii and xcix, where the names are also given. Having already alluded to the chief points in connection with these experiments, whilst describing the various places visited, Concluding remarks. it is not considered necessary to refer to them again. The observations concerning the physical geography of the 77 stations are of a more superficial nature than I could have desired, but the time at my disposal was very limited, and correct information on such matters could not be obtained without personal inspection. It will, indeed, be evident that the experiments referred to in the whole of this report are of an elementary nature. This is, in part, owing to the short period which has elapsed since they were commenced, partly also to my having been tempted, by the desire for results, to keep too many irons in the fire. I trust, however, that what has been done will prove to be a foundation whereupon better things may be built. In conclusion, I respectfully tender my most sincere thanks to Dr. Muir, c. 8., Inspector General of Hospitals, British Troops, for the assistance which he has so gladly rendered on every possible occasion to. further this inquiry, and for the personal interest he has taken, in the details thereof; also to Dr. Cuningham, the Sanitary Commissioner with the Government of India, for similar aid, not lesa cheerfully given. 78