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Chironomid substrate—specific associations regarding the nature (organic—inorganic) and stability (stable—unstable) of different habitats were investigated at two low order Patagonian streams, during high and low water periods. Nant y Fall and Glyn rivers were visited twice (October 2007 and March 2008) and seven different habitat types were identified. A total of 60 samples were collected using a Surber sampler (0.09 m -2 and 250 µm) and a set of 23 environmental descriptors including physicochemical parameters and different fractions of particulate organic matter were assessed. 35 Chironomidae taxa were recorded with Orthocladiinae (20), Chironominae (7), and Podonominae (4) being the most well—represented subfamilies. Paratrichocladius sp. 1, Parapsectrocladius sp. 2, Parametriocnemus sp. 1, Pseudochironomus sp., and Rheotanytarsus sp. were the most abundant taxa. According to the relative preference index, at least 14 taxa showed strong affinity for a particular substrate. The structurally complex macrophyte Myriophyllum quitense supported 11 taxa compared with only five taxa found on the less complex Isoetes savatieri. Generally, stable substrates (boulders, cobbles, and rooted plants) supported significantly higher chironomids richness, abundance, and diversity than unstable ones (gravel—sand). Canonical correspondence analysis revealed that detritus (leaves, seeds, and biomass), macrophyte biomass, and secondarily hydraulic variables had high explanatory power on chironomids species composition and structure. This work suggests that more complex substrates showing persistence in the temporal dimension supported a diverse array of chironomids, meaning that the maintenance of natural habitat heterogeneity is essential for the community. Land—use practices having significant effects on ecological stream attributes such as increased turbidity, sediment deposition, and runoff patterns will alter assemblages. Understanding environmental associations of the Chironomidae assemblage at the habitat scale is significant for conservation purposes and for the management of low order streams in Patagonia.