In this study, we examined the stable fly, Stomoxys calcitrans (L.) (Diptera: Muscidae), larval developmental habitat within the round hay bale feeding sites on cattle pastures, and we identified three zones with distinct characteristics around two types of hay feeders (ring and cone). The parameters monitored in each zone included stable fly emergence, substrate temperature, moisture, pH, thickness of hay-manure layer, and concentration of fecal coliform bacteria (Escherichia coli and Klebsiella oxytoca) as indicators of fecal material. All measurements were conducted during the period of high stable fly prevalence (HSF) in May-June and low stable fly prevalence (LSF) in July-August to better understand the environmental factors influencing stable fly seasonality. Substrate temperature and fecal coliform concentration were the only two significantly different factors between HSF and LSF. Temperatures ranged from 21 to 25°C during HSF versus 25–30°C in LSF but all were within the range for successful stable fly development. Fecal coliform concentrations ranged from 4.2 × 10³ to 4.1 × 10⁴ colony-forming units (CFU)/g of the substrate during HSF and from undetectable (<10) to 100 CFU/g during LSF. Furthermore, we evaluated the effect of different hay:manure ratios (0:1, 1:1, 2:1, and 5:1) on stable fly development (egg to adult). Temperature was significantly higher and stable fly developmental time significantly shorter in all substrates containing hay when compared with that of manure alone, but no significant differences were detected in stable fly emergence among the substrates. These results strongly indicate that the fecal microbial community plays an important role in stable fly larval development in hay feeding sites and that it is the main factor behind stable fly developmental seasonality on pastures. Our results also demonstrate that animal manure mixed with hay provides conditions for faster stable fly development than manure alone; however, hay does not significantly affect overall stable fly emergence.