Three years earlier, in 1887, Ward's attention had been drawn by a happy accident to the physiological aspect of symbiosis, and it never ceased to occupy his mind. It was well known that ginger-beer was made in villages in stone bottles. The fermentation was effected by the so-called "ginger-beer plant" which was passed on from family to family, but nothing was known as to how or where it originated. It seemed to have some analogy with the Kephir of the Caucasus. A specimen was sent to me from the Eastern Counties, and it stood for some time in the sun in my study. I noticed the vigorous growth accompanied by a copious evolution of gas. Ward coming to see me one day, I handed it over to him as a problem worth his attention. At the same time Prof. Bayley Balfour had examined it and concluded that it was a mixture of a yeast and a bacterium. Its study involved Ward in a very laborious research which occupied him for some years, and of which the results were published in the Phil. Trans. in 1892. It proved to be a mixture of very various organisms, every one of which Ward exhaustively studied. This required not less than 2000 separate cultures. The essential components proved to be, as Balfour had suggested, a yeast derived from the sugar and a bacterium from the ginger. Both were anaërobic; the yeast fermented cane-sugar with the copious production of carbon dioxide but little alcohol; the bacterium also produced carbon dioxide, even in a vacuum tube.
The action of the two components studied separately proved to be not the same as when they worked in concert. This was conspicuously the case with the evolution of carbon dioxide, which proceeded with such violence as to make the research attended with considerable danger. It is known that the action of ferments may be checked by the inhibition of the products formed. Ward pointed out that while the use of these might be advantageous to the bacterium, their consequent removal might be equally so to the yeast. This established the important principle of symbiotic fermentation and gave it a rational explanation. On the morphological side Ward showed that the ginger-beer plant is comparable to a gelatinous lichen, and, having resolved it into its constituents, successfully reconstituted it.
The new conception threw a flood of light on many obscure points in fermentation generally, and it is not surprising that Ward's work at once attracted the attention of the brewing industry. It led him to an even more fertile suggestion, that of metabiosis. It was known that the finest wine is sometimes produced from mouldy grapes. He regarded this as a case of one organism preparing the way for another. He returned to the subject in a lecture given at the British Association at Dover in 1899 and pointed out that in the Japanese manufacture of Saké, an Aspergillus prepares the way for the yeast. He also showed that metabiosis played an important part in nitrification.
Fungi cannot draw their nutriment from solid materials without first profoundly modifying them. They accomplish a large part of their digestion, so to speak, externally to themselves. This constantly occupied Ward's mind. He insisted on the part played in the process by ferments. The hyphae of Stereum (Phil. Trans. 1898) delignify the walls of the wood elements of Aesculus layer by layer, and then consume the swollen cellulose. He failed, however, to isolate the ferment which does the work. Nor was he more fortunate with the little known fungus Onygena, which grows on horn, hoofs and hair, setting free ammonia as a final product (Phil. Trans. 1899). That there must be some hydrolysis of keratin can hardly be doubted, for Ward established the remarkable fact that the walls of the hyphae contain no cellulose, but are composed of chitin. Onygena has, in fact, abandoned a plant for an animal nutrition. This would place the germination of the species at a great disadvantage. But he found that this difficulty was overcome by the spores which had been licked from the skin germinating in the gastric juice of the animal's stomach, and, when voided in the excreta, infecting a new host by accidental contact. In the case of both Stereum and Onygena he accomplished for the first time the difficult task of tracing their life-history from spore to fructification.
Ward had prepared himself for the study of bacteria, and in the nineties he undertook, with Prof. Percy Frankland, a prolonged research on behalf of the Royal Society as to the conditions of their occurrence in potable water. The reports of the results fill a thick volume, and the amount of work involved is almost incredible. The bacteriology was entirely due to Ward.
That bacteria are not an inevitable element in potable water is proved by their absence from that of deep springs. They are arrested by filtration through the earth's crust. In any river system they are comparatively fewer towards the watershed, and more frequent towards the mouth. The obvious conclusion is that they are derived from the drainage of the land. As it is known that the bacteria of cholera and typhoid are water-borne, it becomes a problem of vital importance to ascertain if river water is a possible means of distributing these diseases. Ward set to work to ascertain: (i) What was the actual bacterial flora of Thames water; (ii) if this included any pathogenic organisms; (iii) if not, what became of them? The labour required by the first two branches of the enquiry was enormous; he identified and cultivated some eighty species; the resulting answer to the second was happily in the negative.
As to the third, two facts were known. First, that river water, if stored, largely cleared itself of bacteria by mere subsidence; secondly, that Downes and Blunt, in a classical paper communicated to the Royal Society in 1877, had shown that exposure to direct sunlight is fatal to bacteria in a fluid medium. Ward showed that subsidence could not be entirely relied on, as the sediment might easily become the source of re-infection. The effect of sunlight required more critical examination.
It was known that the spores of anthrax were liable to be washed into rivers. Ward determined to study this as the most extreme type of pathogenic infection. As it is undoubtedly the most deadly micro-organism known, and Ward proposed to deal with it on a large scale, it implied no small degree of courage. He found that the spores of anthrax were effectually killed by a few hours' exposure to even the reflected light of a low winter sun. It was clear that this was due to the direct action of the light and not to any heating effect, apart from the fact that they will tolerate boiling for a few minutes. It was further shown that there was no foundation for the theory of Roux and Duclaux that their death was due to poisoning by products of oxidation of the food-medium. Proof of this, indeed, was hardly required, for Pasteur had shown that the bacteria floating in the atmosphere are mostly dead. Were it not so, no surgical operation would be possible. To the bactericidal effect of sunlight is equally to be attributed the absence of bacteria from the High Alps.
The next point was to ascertain to what rays the effect was due. The spores of anthrax are so minute that, when mixed in large numbers with gelatine, they do not affect its transparency, A plate of glass coated with the mixture is at first clear, but ceases to be so if kept in the dark, owing to the germination of the spores. Ward found, in fact, that a photograph could be printed with it, the darkening being the reverse of that of a silver plate. After experiments with coloured screens he completely solved the problem in 1893, with the aid of apparatus supplied by Sir Oliver Lodge and some advice from Sir Gabriel Stokes, by photographing the spectrum on such a plate. It was at once seen that the destructive effect was due to rays of high refrangibility, and, what was extremely important, extended to, and found its maximum in, the ultra-violet. The same results were obtained with the typhoid bacillus. He made the suggestion that the arc light might be used for the disinfection of hospitals and railway carriages.