The physicochemical parameters of the sulphide spring water from a 500 m deep artesian borehole in Sovra Valley (Slovenia) are stable, with very little variation: temperature, 9.2 ± 0.0 °C; pH 7.70 ± 0.11; specific electrical conductivity, 777 ± 8 μS/cm; and dissolved oxygen, 0.19 ± 0.11 mg/l. The water from the borehole is hard, rich in sulphates, and has a notable concentration of sulphide, while it is low in chlorides. Communities of phototrophs were analysed at the oxygenic ecocline at the outflow from the borehole, and from further downstream. Caloneis tenuis, Frustulla vulgaris, Gomphonema sp., Navicula radiosa, Oscillatoria sp., and Tribonema vulgare successfully thrive exclusively at the outflow from the borehole under sub-optimum conditions for the majority of phototrophs. Downstream from the borehole, where the hydrogen sulphide degasses, autotrophic biofilms were dominated by different diatoms. The Bray–Curtis similarity index confirms the distinctive phototrophic communities along the oxygenic ecocline downstream from the sulphidic spring. The sulphidic ecocline downstream from the spring is indicative of the composition of phototrophic communities, as some species appear only downstream from the spring. An ecocline downstream from the borehole of the sulphidic spring was reflected also in the bacterial indicator groups. This study brings novel insights to the limited knowledge in the field of oxygenic phototrophy related to sulphidic springs.