Amoebae interact with bacteria in multifaceted ways. Amoeba predation can serve as a selective pressure for the development of bacterial virulence traits. Bacteria may also adapt to life inside amoebae, resulting in symbiotic relationships. Indeed, particular lineages of obligate bacterial endosymbionts have been found in different amoebae. Here, we screened an extensive collection of Dictyostelium discoideum wild isolates for the presence of these bacterial symbionts using endosymbiont specific PCR primers. We find that these symbionts are surprisingly common, identified in 42% of screened isolates (N = 730). Members of the Chlamydiae phylum are particularly prevalent, occurring in 27% of the amoeba isolated. They are novel and phylogenetically distinct from other Chlamydiae. We also found Amoebophilus symbionts in 8% of screened isolates (N = 730). Antibiotic-cured amoebae behave similarly to their Chlamydiae or Amoebophilus-infected counterparts, suggesting that these endosymbionts do not significantly impact host fitness, at least in the laboratory. We found several natural isolates were co-infected with multiple endosymbionts, with no obvious fitness effect of co-infection under laboratory conditions. The high prevalence and novelty of amoeba endosymbiont clades in the model organism D. discoideum opens the door to future research on the significance and mechanisms of amoeba-symbiont interactions.

Although temperature-correlated shifts in the timing of egg-laying have been documented in numerous bird species, the vast majority of species examined to date have been those that breed in Europe and have an animal-based diet during breeding. However, given that the timing of breeding can be driven, either in the proximate or in the ultimate sense, by seasonal fluctuations in food availability, the relationship between temperature and laying may differ with diet. Here, we report on patterns of reproductive timing in House Finches Haemorhous mexicanus, a North American species that breeds on a primarily seed-based diet. Analysing nest records from House Finches in California spanning more than a century, we found that egg-laying occurred significantly earlier in warmer springs. We also found that although the timing of egg-laying does not show long-term changes in most of California, in the hottest region of the state (the southeast desert basin) it has advanced significantly.

Temperature-correlated shifts in reproductive timing are now well documented in numerous bird species. However, whether temperature directly influences reproductive timing or whether its effects are mediated by an intermediate environmental cue, such as plant phenology, remains poorly understood. In this study, we investigated the direct effects of temperature on reproductive timing in house finches (Haemorhous mexicanus), which have a range and breeding diet not well represented in previous studies of temperature and reproductive timing. We conducted experiments with captive male house finches in which temperature was elevated within realistic ranges and the effects on the timing of preparations for reproduction, as well as on the termination of reproduction and the onset of prebasic feather molt, were examined. We found no adjustments in the timing of reproductive preparations of males in direct response to temperature. However, elevated temperature did advance the breeding-molt transition. Our results suggest that elevated temperatures in the range tested here do not directly impact physiological preparations for reproduction in male house finches, but may constrain the timing of the breeding-molt transition in this species.

Symbiotic associations can allow an organism to acquire novel traits by accessing the genetic repertoire of its partner. In the Dictyostelium discoideum farming symbiosis, certain amoebas (termed "farmers") stably associate with bacterial partners. Farmers can suffer a reproductive cost but also gain beneficial capabilities, such as carriage of bacterial food (proto-farming) and defense against competitors. Farming status previously has been attributed to amoeba genotype, but the role of bacterial partners in its induction has not been examined. Here, we explore the role of bacterial associates in the initiation, maintenance, and phenotypic effects of the farming symbiosis. We demonstrate that two clades of farmer-associated Burkholderia isolates colonize D. discoideum nonfarmers and infectiously endow them with farmer-like characteristics, indicating that Burkholderia symbionts are a major driver of the farming phenomenon. Under food-rich conditions, Burkholderia-colonized amoebas produce fewer spores than uncolonized counterparts, with the severity of this reduction being dependent on the Burkholderia colonizer. However, the induction of food carriage by Burkholderia colonization may be considered a conditionally adaptive trait because it can confer an advantage to the amoeba host when grown in food-limiting conditions. We observed Burkholderia inside and outside colonized D. discoideum spores after fruiting body formation; this observation, together with the ability of Burkholderia to colonize new amoebas, suggests a mixed mode of symbiont transmission. These results change our understanding of the D. discoideum farming symbiosis by establishing that the bacterial partner, Burkholderia, is an important causative agent of the farming phenomenon.