The Greatest Missed Microscopical Opportunity? Paul Bingham

Introduction In this article I explore the proposition of mine who spent time on the Isle of that the greatest missed optical micro- Wight, where I live. scopical opportunity occurred in 1854, Background when Arthur Hill Hassall (AHH) (1817- In the seventeenth century, microscopy 1894) failed to recognise the or- first revealed the presence of micro-or- ganism in specimens that would have ganisms, and in the early nineteenth cen- been teeming with the bacteria; the same tury the contamination of London’s year in which the scientific community drinking water was made public, includ- did not accept the findings of Filippo Pa- ing through a cartoon showing what could cini (1812-1883), who is now credited with be seen down a microscope. Progress, the discovery ofVibrio cholerae. Had, in however, faltered as bacteria are hard to 1854, a majority of the ‘establishment’ ac- visualise in aqueous fluids, being mainly cepted that cholera was caused by an or- water themselves. They generally require ganism, mainly transmitted in water staining or a technique like phase con- rather than ‘bad air’, many lives might trast to make them ‘visible’. In addition, have been spared. (The cholera vibrio was in the 1850s there was a strong belief in brought to general acceptance in 1884 by an alternative theory of disease transmis- (1843-1910)). As with many sion (the miasmic theory of ‘bad air’), that historical retrospectives, the ‘facts’ are favoured anticontagionism and the avoid- contested. To make this article ‘stream- ance of quarantine. The first character- lined’, it is presented initially as a isation of a pathogenic bacterium (an- timeline with a commentary. Seven ‘rows’ thrax), was not complete until the 1860s, from the timeline, termed Exhibits, have and staining of bacteria was not perfected then been selected for more detailed con- until the late 1870s/early 1880s. sideration. The timeline places perhaps undue emphasis on AHH, but he is a hero

The Timeline

Date Event/publication Commentary (Exhibit)

Leeuwenhoek: communicated from First microscopical demonstration of the existence c1670 Holland with the Royal Society of micro-organisms.

Leeuwenhoek: said to be first to use c1673 Extracted dye from the saffron crocus bulb. a stain in microscopy

1828 William Heath: Cartoon ‘Monster Early attempt to make the public aware of what Soup’ - depicting contamination in could be seen down the microscope. (Exh 1) water taken from the Thames

The first book in English on histology. Helped AHH: ‘Microscopic Anatomy of the 1846 establish AHH as one of the leading microscopists Body in Heath and Disease’ of his day.

1 Date Event/publication Commentary (Exhibit) These microscopical findings were discred- William Budd: Letter to The Times 1849 ited by the Royal College of Physicians and other communications; claimed (correctly) and are likely to have made he had seen ‘fungous cells’ in microscopists wary of making subsequent (Exh 2) association with cholera fluids claims of having seen the cholera organism. 1850 AHH: ‘Microscopical examination of Drawings of contamination seen under the the water supplied to the inhabitants microscope in samples of water supplied by (Exh 3) of London etc’ various London water companies. Hand washing has general application to all Ignaz Semmelweiss: Advocated infectious diseases including cholera. Essential 1850 washing hands to prevent the spread for those caring for cholera patients and of childbed fever handling cholera pathology samples! Pollender, Rayer & Davaine: Characterised the anthrax bacillus under the 1850-68 ‘Discovered’ the anthrax bacillus microscope. Snow argued that cholera was spread mainly 1854 : ‘On the Mode of by contaminated water, but also by touch and Communication of Cholera’ (2nd in food. At the time this was not generally (Exh 4) much enlarged edition) accepted: spread by bad air was widely favoured. 1854 Filippo Pacini: ‘Microscopical Described the comma-shaped cholera observations and pathological organism. Published in Italian but noted by (Exh 5) deductions on cholera’ Farr. Recognised in 1965 – see below. Lionel Beale: ‘The Microscope in 1st ed – envisaged only very limited role in 1854 Medicine’ medicine. Bacteria not mentioned.

AHH pioneer of detection, using the AHH: ‘Food its adulterations & 1855 microscope, of what became ‘criminal’ methods for their detection’ adulteration of food and drugs.

Part of a Report to both Houses of Parlia- 1855 AHH: ‘Report on the microscopical ment on the Cholera Epidemic of 1854. examination of Blood and Excretions, Perhaps surprisingly, AHH did not suggest that (Exh 6) &c of Cholera Patients’ the vibriones he saw were the cause of cholera.

1863 AHH: ‘Urine in health and disease’ AHH published widely on microscopy.

William Farr (lead civil servant on By 1866 Farr concluded that cholera was 1866 medical statistics): Report on the transmitted by ‘cholera flux’, mainly in 1866 outbreak of cholera contaminated water, and not bad air.

Farr praised mathematical work Pacini had undertaken on cholera. Farr did not, however, Farr: visit to Italy where he met with 1867 endorse Pacini as the discoverer of the cholera Pacini organism - a further missed microscopical opportunity!

Karl Weigert: alcohol-methylene blue Weigert introduced into bacteriology, staining 1875 staining of bacteria techniques in use for histology.

Robert Koch: performed a series of The first time a microorganism was experiments on anthrax fulfilling what 1876 conclusively linked with a specific disease. First came to be known as Koch’s ‘proof’ of the Germ Theory. postulates

2 Date Event/publication Commentary (Exhibit) Robert Koch: Photograph ofBacillus 1877 First photomicrograph of a bacterium. anthracis Paul Ehrlich: developed an ‘acid-fast In 1885, technique finalised by Ziehl & 1882 stain’. Neelsen. Previously, Jean-Antoine Villemin had shown Robert Koch: announced the discovery of that TB was transmissible. Koch advanced 1882 the main organism causing tuberculosis. several lines of evidence to show his 1905, awarded a Nobel Prize identified organism is the cause of TB. Later refined into ‘Koch’s Postulates’. Hans Christian Gram: published his 1884 Devised Gram stain in 1882. famous stain Again, Koch advanced lines of evidence, but Robert Koch: announced the discovery of at that time, could not show that his cholera 1884 the bacterium . 1965 organism caused disease in any ‘lower organism ‘reassigned’ animal’. Correspondents argued that Pacini should be Correspondence about the discovery of credited with the discovery of the cholera 1884 the cholera vibrio in the Lancet, including organism. AHH maintained he saw the letters from AHH organism before Pacini.

International committee on nomenclature: 1965 adopts Vibrio cholerae Pacini 1854 as the Decision reached largely due to the urging of (Exh 7) correct name of the cholera causing Prof Rudolph Hugh of USA. organism.

Exhibit 1: Cartoon ‘Monster itan Water Supply. The growing popula- Soup’ by William Heath tion and increased use of drains for dis- posing of sewage (rather than cesspits and carting manure to fields), was polluting the River Thames, from which water companies ex- tracted their supply. The cartoon depicts a fashionable lady viewing a sample of drink- ing water through a microscope and dropping her cup of tea in re- sponse to what she saw. The cap- tion at the top of the frame reads: Microcosm, dedicated to the Lon- don Water Companies, Brought Forth All Monstrous, All Prodi- gious Thigs [sic], Hydras, and Gorgons, and Chimeras Dire. Vide Milton [Paradise Lost]. Fig. 1: Cartoon Monster Soup by William Heath Bottom of the frame: In 1828, the caricaturist William Heath Monster Soup Commonly Called published a cartoon lampooning the qual- Thames Water, Being a Correct Rep- ity of water being supplied by London wa- resentation of that Precious Stuff ter companies. This coincided with the Doled Out To Us!!! 1827-28 Royal Commission on Metropol- 3 Exhibit 2: William Budd: described microscopic organisms he claimed to be the cause of cholera In 1849, when cholera was affecting Bris- tol, the local medical society appointed a microscopical committee including Fred- erick Brittan, Joseph Swayne and Wil- liam Budd. Subsequently, they announced widely (example below) they had found the cholera-causing organism. However, these findings were not suppor- ted by the Royal College of Physicians. The organism ‘discovered’ was probably a fungus and definitely not the cholera vi- brio’. London Journal of Medicine, 1849, page 987. Discovery of peculiar living organisms Figure 2: AHH’s depiction of organic matter in the ‘’Rice-water’’ evacuations of concentrated from drinking water supplied by Cholera’. ‘Drs J. G. Swayne and Brit- the New River Company tan, of Bristol, having been for some time engaged, under the direction of the Medico-Chirurgical Society of that their discoverer; most of these ‘eponym- city, in the microscopic investigation ous titles’ have subsequently fallen into of subjects connected with cholera, disuse.) state that they have discovered, in the rice-water discharges, and also in the In 1850, as one of his first publications on atmosphere of infected places, certain ‘adulteration’ (defined in this instance as: living organisms, to which they are ‘making profit by supplying contaminated disposed to attach much importance products, in extreme cases adding in the etiology of cholera. Dr. William cheaper ingredients, toxic colourants etc Budd, of Bristol, in a letter to the to make a product go further or change its Times of September 26, confirms their appearance to make it more saleable’), statements, and mentions that he has AHH published a microscopical review of detected similar organisms in water the piped water supply sold to London in- procured from cholera districts. habitants by various privately owned wa- ter companies. The review revealed Exhibit 3: ‘A microscopical exam- extensive pollution, indicating no im- ination of the water supplied to provement since William Heath’s cartoon the inhabitants of London and ‘Monster Soup’. the suburban districts’ by Dr It is difficult to assess the impact of Hassall AHH’s 1850 publication, which he pre- Arthur Hill Hassall qualified as a medical viewed in the Lancet. In 1851, AHH gave doctor, and early in his career became evidence to a Parliamentary Committee well known as an expert in the applica- of Inquiry on the water supplies of the tion of the microscope to natural history, Metropolis. The 1852 Metropolis Water writing books on freshwater algae, ana- Act resulted, but change was slow to fol- tomy, pathology, and public health, espe- low and, in the end, real improvement de- cially on the detection of the adulteration pended on Bazalgette’s sewage engin- of drugs and food. Early in his career, eering scheme that was opened in 1865. Hassall discovered microscopic features Prior to that, in 1854, AHH undertook a in the thymus gland that are still widely further review of the microscopy of Lon- known as ‘Hassall’s Corpuscles’, rather don’s water supply (see Exhibit 5). than ‘Thymic Corpuscles’. (In the nine- Exhibit 4: ‘On the Mode of teenth century many anatomical and his- tological new findings were named after Communication of Cholera’ by

4 John Snow (2nd enlarged what ever their local reputation may edition) be. 4a) Involvement of Hassall by Snow Page 109: in the Broad Street pump outbreak It must not be disguised, however, Page 52: that medical men are not yet generally convinced that the disease is actually Hassall, who was good enough to ex- communicated from person to person amine some of this water with the mi- by the morbid matter being swallowed croscope, informed me that these in the drinking water, or otherwise. particles had no organised structure, and that he thought they probably res- John Snow is the modern hero of cholera, ulted from decomposition of other and has his own Society (johnsnowsociety matter. He found a great number of .org), because he got it right; cholera is very minute oval animalcules in the mainly spread by contaminated water. He water, which are of no importance, ex- was disregarded in his day because the cept as an additional proof that the medical elite favoured an alternative hy- water contained organic matter on pothesis (cholera transmitted by ‘bad which they lived. The water also con- air’). Snow was not himself a member of tained a large quantity of chlorides, the medical elite, (although he gave chlo- indicating, no doubt, the impure roform to Queen Victoria in labour), and sources from which the spring is sup- his methods and conclusions were cri- plied. Mr. Eley, the percussion-cap tiqued in detail - Snow did make mistakes manufacturer of 37 Broad Street, in- (Parks 1854). formed me that he had long noticed Exhibit 5: ‘Microscopical that the water became offensive, both observations and pathological to the smell and taste, after it had deductions on cholera’ by been kept about two days. This, as I Filippo Pacini noticed before, is a character of water contaminated with sewage. Another Like Hassall, Filippo Pacini qualified as a person had noticed for months that a medical doctor and became skilled in the film formed on the surface of the wa- growing application of the microscope. By ter when it had been kept a few hours. contrast, Pacini was university based, and became Professor of Anatomy at 4b) Snow’s references to ‘morbid Florence in Italy. Early in his career, Pa- matter’ cini described a tiny nerve sensory ending (Snow, along with other practitioners of that is found in skin and other tissues. the time, was a follower of the ‘Zymotic This became widely known, and to this theory’, a term that had been coined by day generally retains its eponymous la- William Farr (1807-1883).) bel, ‘Pacinian Corpuscle’. Page 15: In 1854, when Florence was affected by cholera, Pacini was able to perform only a For the morbid matter of cholera hav- limited number of post-mortems on chol- ing the property of reproducing its era patients, but conducted histological own kind, must necessarily have some examinations of their intestinal mucosa. sort of structure, most likely that of a Pacini published a paper that was sub- cell. It is no objection to this view that sequently key, in 1965, to the decision of the structure of cholera poison cannot the International Committee on Nomen- be recognised by the microscope, for clature. However, at the time an anonym- the matter of smallpox and of chancre ous reviewer, published in The British can only be recognised by their effects and Foreign Medico-Chirurgical Review and not by their physical properties. 16: 144-145, stated: Page 54: The Florentine micrographer prefaces We must conclude from this outbreak his memoir with the statement, that that the quantity of morbid matter the results recorded in it are few and which is sufficient to produce cholera incomplete, owing to the smallness of is inconceivably small, and that the his opportunities for the practical shallow-wells in a town cannot be study of pathological anatomy. looked on with too much suspicion, 5 Four cases of Asiatic cholera were the qualified microscopist, was commissioned bases of these observations. In the only by the government (Board of Health) dur- two cases in which he examined the ing the course of the 1854 cholera epi- vomited matter it contained sarcinae demic to produce two reports, the first on ventriculi; in one he additionally microscopy of the public water supply (to found some vibrios of the genus Bank- an extent replicating work he had under- sium. Only once did he examine the taken in 1850 (exhibit 3)) and the second faeces, and then without particular on the microscopy of rice water diarrhoea, result. In three of the cases he ex- blood, urine etc., from cholera patients. amined the intestinal fluid, and found Only the second of Hassall’s reports is fea- floating in it a large quantity of epi- tured here, and then only the microscopy thelium, and some detached villi. In of rice water evacuations. one of the cases he additionally found Hassall related that he had examined many of the choleraic fungi, and an samples from 25 patients including ma- immense number of vibrios. On ex- terial that was fresh (passed within 2 amining the corresponding mucous hours of examination), and a sample ob- membrane, it appeared anaemic, and tained at post-mortem from the small in- completely stripped of its epithelial testine. Hassall found ‘myriads of covering; this condition, by permitting vibriones’ in all of these samples. Hassall the extraordinary serous extravasa- noted: tion, he regards as the first and prin- cipal pathological condition of • Of these vibriones many formed cholera. To develop this proposition, threads more or less twisted, while our author engages in lengthened ar- others were aggregated into masses, guments, which however ingenious, which under the microscope, presen- have failed to remove from our mind ted a dotted appearance. an impression of regret that before • It thus appears that vibriones are their publication a much larger num- constantly present in the rice-water ber of observations was not instituted, discharge of cholera, and that they and the micrographer's power of are developed in it during life, and judging their value increased by his while still retained in the small in- studying the disease en clinicier. We testines. cannot but think that, in studying cholera, the renowned discoverer ‘’dei • It is possible that they may obtain en- nuovi organi" if he has not actually trance into the stomach and bowels mistaken an effect for a cause, has by means of the atmosphere, and it is raised an incident to the unmerited perfectly certain that they do fre- position of an essential and funda- quently gain admission through some mental feature. Pacini was unable at the time to convince his peers or the au- thorities that he had dis- covered an organism that was associated with, and the cause of, cholera. In 1867, he was un- able to convince William Farr when he visited Florence, in spite of Farr’s recent recogni- tion that cholera was transmit- ted by contaminated water. Exhibit 6: ‘Report on the microscopical examina- tion of Blood and Excre- tions, &c of Cholera Patients’ by Dr Hassall Figure 3: AHH's 1855 depiction of 'Rice water evacuation Arthur Hassall, who had be- of cholera'. (d) Acicular crystals (e) Fragments of muscular come an established, medically fiber (f) Vibriones (in a chain) (g) Masses of vibriones

6 of the impure waters consumed, in suggested there was a primary connection which I have not unfrequently detec- between the vibrions and cholera? ted the presence of vibriones, some- times in considerable numbers. Exhibit 7: International committee on nomenclature: • Once introduced into the alimentary Adopts Vibrio cholerae Pacini canal, they are brought into relation 1854 as the correct name of the with conditions highly favourable to their development and propagation, cholera causing organism. both of which take place with almost In his ‘Request for an opinion’ to the In- inconceivable rapidity. ternational Committee on Nomenclature, Prof Rudolph Hugh referred to the 1854 • I have made two or three examina- paper by Filippo Pacini and concluded: tions of healthy and natural faecal evacuations at the time of their being Pacini saw the curved etiologic agent passed, and in these I have detected of Asiatic cholera in tremendous num- the presence of comparatively a very bers in denuded intestinal epithelium small number of vibriones. and intestinal contents of victims. The pathological changes in the intestine In 1855, AHH concluded, in retrospect and the organism itself were described surprisingly: in such a manner that they may be re- Without, however, at all supposing cognised. that there is any essential or primary The International Committee, with a connexion between these vibriones and hundred years’ experience of the germ cholera, their occurrence in such vast theory, sided with Pacini and the name of numbers in the rice-water discharges the vibrio was changed from Vibrio chol- of that disease is not without interest, erae Koch 1884 toVibrio cholerae Pacini and possibly is of importance. 1854. In 1884, following the announcement of Conclusion Koch’s discovery, Hassall wrote three times to the Lancet and was at pains to Was 1854 a credible opportunity for underline his belief that he saw vibriones the visualisation of the cholera before Pacini. He then included the sen- vibrio and its demonstration to the tence: medical establishment? I would remark, however, that while I Factors mitigating against: recognise the importance of this dis- • William Budd’s 1849 announcement covery I did not go to the length of de- of the discovery of the cholera organ- scribing it as the cause of cholera. ism had been discredited. But that is the point, Pacini consistently • By 1854, no bacterial organism had claimed that what he had seen was the been discovered and no staining of cause of cholera – even though he was not bacteria had taken place. generally believed. • Snow’s contention that cholera was In his autobiography published in 1893, mainly spread by contaminated AHH noted drinking water and caused by ‘mor- … from the characters exhibited and bid matter’, that he believed was an which are fairly represented in the fig- organism, had not been generally ac- ure (reproduced above), and from cepted by the medical establishment their general agreement with Koch’s Factors mitigating for description, there is not the smallest doubt that the cholera bacillus was • AHH (along with others) was an ex- present in the discharges in nearly pert of microscopy whose opinion was every case and was first seen by me likely to have been respected. during the cholera epidemic of 1854, • AHH described and illustrated ‘vibri- now nearly 40 years since. ones’, that in retrospect were the Although AHH reflected on many things cholera vibrio. in his autobiography, he did not reflect on • AHH knew Snow personally, and is the elephant in the room, why had he not likely to have been well acquainted with his ideas, but for some reason he 7 Fig. 4: Hassall’s corpuscle Fig. 5: Pacinian corpuscle

did not integrate Snow’s ideas with his vibriones. Lionel Beale 1854 ‘The Microscope in Arguably Pacini had less opportunity (few Medicine.’ Digital copy accessible free via cases of cholera to study and he was not archive.com government sponsored) than AHH. In the Filippo Pacini 1854 ‘Microscopical obser- view of a small number of contemporary vations and pathological deductions on supporters, and in retrospect, the Inter- cholera.’ Original in Italian. Translation national Committee on Nomenclature, he into English not published. not only described the cholera vibrio but hypothesised it was the cause of cholera. John Snow 1854 ‘On the Mode of Com- At the time, however, he was not gener- munication of Cholera.’ Digital copy ac- ally believed. cessible free via archive.com Should Cholera vibrio have been General Board of Health 1855 ‘Appendix renamed? to report of the committee for scientific inquiries in relation to the cholera- Pacini may have been the first to ‘dis- epidemic of 1854’. Digital copy accessible cover’ the cholera vibrio, but that made no free via archive.com apparent difference. Robert Koch appears initially to have believed that he was first EA Parks 1855 ‘Dr Snow on the Commu- to have discovered the vibrio. Although nication of Cholera.’ British and Foreign change was slow, his ‘discovery’ did make Medico-Chirurgical Review, 15: 449-63. a difference. Digital copy accessible free via National Centre for Biotechnology Information. Are there other contenders for the greatest missed optical microscop- AHH 1893 ‘Narrative of a Busy Life: An ical opportunity? Autobiography’. Digital copy accessible free via archive.com For this question, it is over to readers for as many nominations as possible. If there Rudolph Hugh 1964 ‘The proposed con- were to be enough interest, PMS could servation of the generic name Vibrio Pa- take a vote! cini 1854.’ Digital copy accessible free via microbiologyresearch.org References AHH 1846 ‘Microscopic Anatomy of the John Eyler 2001 ‘The changing assess- Body in Health and Disease.’ Digital copy ments of John Snow’s and William Farr’s accessible free via archive.com cholera stuidies.’ Digital copy accessible free via electronic ‘John Snow Archive AHH 1850 ‘Microscopical examination of Research Companion’. the water supplied to the inhabitants of https://johnsnow.matrix.msu.edu/ London etc.’ Digital copy unfortunately not available

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