It is apparent by the confirmation of viremia in 2 out of 126 sparrows (Mitamura) that birds get infected with Japanese B encephalitis (JBE) virus in the form of inapparent infection. This can also be presumed by the results of positive neutralization test against JBE virus in some birds. Hammon considered, through neutralization test, that St. Louis encephalitis (SLE) might be widely spread in nature. Smith stated that SLE virus is found among chicken mites (Dermanyssus gallinae) . Encephalitis virus is recognized to spread among birds without mosquitoes. In other words, birds being the original virus reservoir, man and animal get occasional infection with encephalitis virus in the mode of bird-mosquito-human cycle transmission, and birds are playing an important role as a source of infection. Regarding Russian spring-summer encephalitis (RSSE), Solobiev discovered that siskin and finch are susceptible to it. Concerning the epidemiology of encephalitis, Mitamura suspected the existence of some relationship between JBE and birds, and further advanced into setting up a hypothesis that JBE might be tfansmitted from one area to another through migratory birds. Experimental viremia on wild birds has also been carried out by Kobayashi and Kawashima in Japan. Still, the relationship between birds and encephalitis necessitates further investigations. Experiments were carried out from three angles.
(1) Onset and duration of viremia when birds are experimentally infected.
(2) Whether birds caught in fields develop viremia or preserve virus in brain spontaneously or not.
(3) Frequency and extent of the presence of neutralizing and complement fixing antibodies in the blood of birds in situ.

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There are numerous reports on the studies of the flocculation between tetanus toxin and antitoxin, but only a few of them are successful, because this flocculation is disturbed by many other antigen-antibody reactions unlike the case of diphtheria. Recently, Goldie concluded that “by selecting, diluting, blending and heating certain lots of refined tetanus antitoxin, it is possible to prepare a standard tetanus antitoxin giving only a single and specific flocculation zone”. Moloney reported that it is also possible to obtain a standard antitoxin giving single zone by absorbing the peptic digested tetanus antitoxin with both heat-stable and heat-labile non-toxic antigens.
According to Mueller and Miller's report, if a highly potent toxin is used as antigen, the initial flocculation zone will show true toxin-antitoxin reaction. They also suggested that it would be possible to titrate the antitoxin during the immunization process of horse for antitoxin production. In authours' experience, it was also possible to titrate tetanus antitoxins and toxoids with the reference standard antitoxin which was absorbed by Moloney's method. Though the titration of antitoxins by flocculation was rather difficult, even when highly potent antigen was used, it was possible to obtain good results by selecting suitable antigen.
The purpose of this paper is to titrate tetanus antitoxin by direct flocculation method, and to study the relationship between animal test and flocculation test for practical purpose.

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As reported in the first report of the present series, α-glucosidase of Aerobacter cloacae etc participates in the fermentation of sucrose. In the absence of α-glucosidase, however, if fructosidase is present, sucrose will be hydrolyzed into fructose and glucose. Emil Fischer et al proved the above facts by using yeast and such fermentation is called as indirect fermentation. While, it has been insisted that the disaccharide such as sucrose or maltose can be fermented without the process of hydrolysis and numerous studies based on different standpoints have been reported on this problem.
Regarding the sucrose fermentation of living A. cloacae and Esch. coli var. communior etc., as reported before, the oxygen bridge at the α-glucoside conjunction of sucrose is hydrolyzed by α-glucosidase in the former, while, in the latter, as it has no α-glucosidase, the fermentation either by fructosidase or of some other type must take place. It is considered, therefore, that the enzyme participating in the fermentation of sucrose is entirely different between A. cloacae and Esch. coli var. communior. Experiments were carried out in order to know the difference of sucrose fermentation between those two strains, and some results, which indicated the presence of direct fermentation in the sucrose fermentation of Esch. coli communior, have been obtained.

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