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Abstract As DNA sequencing technology continues to rapidly improve, studies investigating the microbial communities of host organisms (i.e., microbiota) are becoming not only more popular but also more financially accessible. Across many taxa, microbiomes can have important impacts on organismal health and fitness. To evaluate the microbial community composition of a particular microbiome, microbial DNA must be successfully extracted. Fecal samples are often easy to collect and are a good source of gut microbial DNA. Additionally, interest in the avian preen gland microbiome is rapidly growing, due to the importance of preen oil for many aspects of avian life. Microbial DNA extractions from avian fecal and preen oil samples present multiple challenges, however. Here, we describe a modified PrepMan Ultra Sample Preparation Reagent microbial DNA extraction method that is less expensive than other commonly used methodologies and is highly effective for both fecal and preen oil samples collected from a broad range of avian species. We expect our method will facilitate microbial DNA extractions from multiple avian microbiome reservoirs, which have previously proved difficult and expensive. Our method therefore increases the feasibility of future studies of avian host microbiomes. 

Abstract Understanding how mating cues promote reproductive isolation upon secondary contact is important in describing the speciation process in animals. Divergent chemical cues have been shown to act in reproductive isolation across many animal taxa. However, such cues have been overlooked in avian speciation, particularly in passerines, in favor of more traditional signals such as song and plumage. Here, we aim to test the potential for odor to act as a mate choice cue, and therefore contribute to premating reproductive isolation between the black‐capped (Poecile atricapillus) and Carolina chickadee (P. carolinensis) in eastern Pennsylvania hybrid zone populations. Using gas chromatography–mass spectrometry, we document significant species differences in uropygial gland oil chemistry, especially in the ratio of ester to nonester compounds. We also show significant preferences for conspecific over heterospecific odor cues in wild chickadees using a Y‐maze design. Our results suggest that odor may be an overlooked but important mating cue in these chickadees, potentially promoting premating reproductive isolation. We further discuss several promising avenues for future research in songbird olfactory communication and speciation. 

Olfaction is an important sense that has contributed to reproductive isolation and speciation in many taxa. However, in birds, olfaction and its potential role in communication has historically been neglected. Thus, what role olfaction plays in avian speciation is mostly unknown. Here, we aim to guide future research by highlighting the likely potential for olfaction to contribute to reproductive isolation and speciation in birds. First, we detail the best-understood example found thus far that is consistent with reproductive isolation by olfaction in birds: black-capped and Carolina chickadees. These species show interspecific differences in preen oil chemistry and conspecific odour preferences within their natural hybrid zone. We then suggest a number of promising avenues for future research and the kinds of systems, approaches and evidence that would help to advance this severely understudied area. In sum, the growing body of research into the chemical ecology of birds suggests an important role of olfaction in many areas of avian life. Future research will be necessary to determine to what extent olfaction contributes to the formation and maintenance of species boundaries in birds.