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Four different associations between ciliates of the family Euplotidae and bacteria have been studied. The ectosymbiotic relationship between ciliates of the genus Euplotidium and epixenosomes, (bacteria related to Verrucomicrobia), offers cues for ecological and evolutionary studies. Epixenosomes, although not vital for their host, confer on them a selective advantage, i.e. defense against predation. Epixenosomes have a compartmentalized cell and possess microtubule-like structures. Interestingly, the presence of two tubulin-like genes has been reported in members of the free-living verrucomicrobial genus Prosthecobacter. We obtained evidence that well-known Polynucleobacter bacteria, upon which a group of fresh-water Euplotes species depends, interfere with the glycogen metabolism of their hosts. Euplotes magicirratus is no longer able to digest food-organisms and is destined to die, once deprived of its specific prokaryotic endosymbiont, likely a new species of the alpha-proteobacterial genus Devosia. A further intriguing finding is the regular presence of rickettsiae in the cytoplasm of a marine Diophrys species. This is the first report of these bacteria in protozoa. This short list suggests that specialized mutualism is more widespread in the phylum than so far supposed.

A symbiotic relationship between the marine ciliate Euplotes magnicirratus and an alpha-proteobacterium has recently been described. The bacterial symbiont, identified by molecular analysis as a new species of the genus Devosia, is referred to with the provisional name Candidatus Devosia euplotis'. This association is constant (it was found in different ciliate strains sampled in different geographical regions), and durable (it persisted in laboratory conditions for years). The physiological significance of this association was investigated through antibiotically produced aposymbiotic E. magnicirratus and attempts to grow the endosymbionts outside their host. Aposymbiotic protozoa completely lost the ability to digest the green alga Dunaliella tertiolecta, their usual food in laboratory conditions, and died. An investigation into the different steps of the digestive process showed that acidification of the digestive vacuoles occurred normally in both symbiotic and aposymbiotic ciliates, but that subsequent activation of acid phosphatase did not occur in aposymbiotic cells. Therefore, the ciliate depends on the symbiont for digestion of its green algal food and, as a consequence, for its survival. Attempts to grow bacterial endosymbionts on diluted culture media and on ciliate homogenate were unsuccessful. The results indicate that this represents a new type of symbiosis between prokaryotes and ciliates, which is mutualistic, and most likely obligatory for both partners.