Experiências foram realizadas com bactérias dos gêneros Listeria e Erysipelothrix, em meios líquido e sólido, utilizando o clorêto de 2, 3, 5 - trifeniltetrazólio. A atividade enzimática redutora das listérias para o TTC foi diferente da do E. rhusiopathiae, principalmente em meio líquido e nas horas iniciais de observação. Preparações feitas para microscopia ótica e electrônica dos germes tratados com TTC revelaram a presença de granulações polares, bipolares e centrais dentro do corpo das listérias. A evidenciação de granulações coradas de formazana, intracelulares nas listérias, confirma estudos anteriores quanto à possibilidade da existência de mitocôndrias nas bactérias.

Based on a similarity of cultural and biochemical properties, WILSON and MILLES (6) compared the microorganisms of listeriosis with those of swine erysipelas including them in the genus Erysipelothrix with the species E. monocytogenes and E. rhusiopathiae. However, BARBER (1) and JULIANELLE (4), in a comparative study of both microorganisms, do not admit that possibility. Observing the morphological and citochemical characteristics of L. monocytogenes (N.º 7 973, 5 348, 5 105, 5 214, of the Seeliger collection) and E. ehusiopathiae cultures (N.º 1, 7, 11, 27, 37, of the WIX collection), both in phase "S", we were able to verify their comparative behavior when in contact with 2-3-5-triphenyltetrazolium chloride (TTC), "Synthetical Laboratories, Chicago, U.S.A". 1. To 1,0 ml of cultures of both microorganisms grown in plain broth plus 5% horse serum for 24 hours at 37ºC, 0.1 ml of sterilized 1% aqueous solution of TTC was added. The tubes were left at room temperature (± 25ºC) or in the incubator (37ºC). In the tubes with L. monocytogenes immediately appeared a red coloration (fig. 1) characteristic of formazan production, that became gradually more pronounced, from bottom to top (figs. 2, 3, 4, 5); at the and of 24 hours the color was intense the tube. In the tubes with E. rhusiopathiae (figs. 1, 2, 3, 4), only a faint pink coloration was seen after 2 hours; at the end of 24 hours, the intensity of color was not equal to that observed in tubes with L. monocytogenes. 2. Petri dishes with 15 ml of plain agar with 5% horse serum, containing 0.1 ml of 1% sterilized aqueous solution of TTC, were inoculated with L. monocytogenes and E. rhusiopathiae, and incubated for 24 hours at 37ºC. Colonies of L. monocytogenes that developed presented a red color, indicative of formazan, as previously described by GRAY et al. (3), while the E. rhusiopathiae colonies did not show any coloration. 3. Disks of filter paper, previously soaked with TTC and dried were placed over the growth of L. monocytogenes in Petri dishes with plain agar plus 5% horse serum form 24 hours. No change in the color of the colonies was observed. The same negative result was obtained when the porous clay cylinder technique for measuring antibiotic activity was employed, With E. rhusiopathiae also negative results were obtained with the disk technique. 4. Wet preparations of the cultures after 1 hour of contact with TTC in the same conditions of item 1, were observed at 900X magnification. L. monocytogenes presented normal shape and dimensions, but one to three intracellular polar, bipolar and or central, red-purple granules were noted. E. rhusiopathiae bacilli were morphologically normal…

Zonas citoplasmáticas de atividade redutora de TTC foram evidenciadas em células de Proteus vulgaris examinadas ao microscópio eletrõnico. As experiências foram realizadas em vários intervalos de tempo, sendo a reação, em alguns casos, estabilizada por meio de formol. Verificou-se que as granulações de formazana podiam ser removidas no interior do corpo bacteriano por meio de tratamento com acetona. O bombardeamento pelos eléctrons, quando os germes não tinham sido fixados com formol, determinou a saída das granulações do interior dos bacilos, deixando vestígios de sua presença (rompimento das membranas celulares nos pontos onde estavam localizados). A fixação com formol evitava a saída dos grânulos. O acúmulo dos cristais de formazana, internamente, nos pólos ou ao longo do corpo bacteriano, provoca sérias alterações morfológicas.

Cells of P. vulgaris incubated in plain broth (4 hours at 37 C) were treated with TCC in final concentration of 0,1 per cent at 37 C. They presented sites of TTC reduction as granules with volumes proportional to the time of contact with TTC. In microorganisms not fixed with formaldhyde, most of the granules during the electron microscopy ruptured the bacterial membranes leaving remains of these, like "ghosts", in the sites where the granule had been located: in rare cases large granules still were seen deforming the bacilli, whit a crystalline contour. When the cells were fixed with formaldehyde the granules remained in the microorganims. They could be removed from the cells by treatment with acetone leaving empty collapsed membranes.