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Symbiotic microbes may mediate songbird chemical signals
Danielle Whittaker
Kevin Theis
Photo by Marine Drouilly
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Symbiotic hypothesis for chemical communication
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Symbiotic microbes in spotted hyena scent pouches are responsible for odors in scent marks
-0.6 -0.5 -0.4 -0.3 -0.2 -0.1 0 0.1 0.2 0.3-0.2
-0.15
-0.1
-0.05
0
0.05
0.1
0.15
0.2
0.25
Structure (Bray-Curtis)
ANOSIM: R = 0.36, P = 0.002
Fig Tree
SouthernComfort
Mara River
Emarti Hill
Bacterial community structure varies with clan membership
(Theis et al. 2012)
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Preen oil is an odor source in birds
Oil secreted from the uropygial gland contains volatile compounds that give birds an odor
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Photo by Danielle Whittaker
Volatile compounds in preen oil vary with:
Species
Sex
Population
Relatedness
Breeding condition
Quality …and could play a role in mate choice.
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Photo by Marine Drouilly
Do avian preen glands harbor odor-producing bacteria?
Do adults transmit these bacteria to their offspring during the nestling phase?
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Dark-eyed junco (Junco hyemalis)Mountain Lake Biological Station, VA
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Dark-eyed Juncos
Seasonal breeders
Socially monogamous
~30% extra-pair fertilization (EPF) rate
Females incubate eggs, both males and females feed nestlings
Nestlings fledge at day 12
Photo by Marine Drouilly
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Photo by Dawn O’Neal
Field Methods
13 nests, 64 juncos
Swab samples from preen glands:• all nestlings (2-4,
mean = 2.9) at age 11-12 days
• both parents at all but 2 nests
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Photo by Marine Drouilly
Extracted DNA from swabs using MO BIO PowerSoil Kits
Illumina MiSeq platform, targeting the V4 region of bacterial 16S rRNA gene
Processed using mothur
Each sample subsampled to 5000 sequences
Bacterial sequences clustered based on 97% nucleotide similarity to define OTUs
Sequencing Methods
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Photo by Flickr user Super Bay
Bacterial communities in junco preen glands have very high levels of diversity
Top 20 OTUs account for ~45% of sequences, with no OTU accounting for more than 5%
In hyena scent glands, the top 20 OTUs account for ~90% of sequences, with the top OTU accounting for ~45%
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Top 20 OTUs in junco preen glands:
Burkholderiaceae, Burkholderia
Burkholderiaceae, Ralstonia
Clostridiaceae, Clostridium
Clostridiales Family XI, Incertae Sedis XI, Anaerococcus
Comamonadaceae, unknown
Enterobacteriaceae, unknown
Enterobacteriaceae, unknown
Enterococcaceae, Enterococcus
Halomonadaceae, Kushneria
Halomonadaceae, Salinicola
Kineosporiaceae, Kineococcus
Micrococcaceae, unknown
Moraxellaceae, Alkanindiges
Moraxellaceae, unknown
Moraxellaceae, unknown
Unknown, unknown
Pseudomonadaceae, Pseudomonas
Rhodobacteraceae, unknown
Sphingomonadaceae, Sphingomonas
Sphingomonadaceae, Sphingomonas
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Other prominent OTUs associated with odor:
Corynebacterium: human axillary odor
Porphyromonas: malodorous breath
Bacteroides, Finegoldia & Fusobacterium: common volatile fatty acid producers associated with many vertebrates
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Bacterial community structure varied by nest
Nests significantly different (70/78 pairwise comparisons)
NPMANOVA, all: F = 3.751, P = 0.0001 (Bray-Curtis)
Nestlings only: F = 6.946, P = 0.0001
Photo by Nicole Gerlach
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all individuals, NPMANOVA, F = 3.751, P = 0.0001
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nestlings only, NPMANOVA, F = 6.946, P = 0.0001
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Nestlings were more similar to mother than father
Wilcoxon’s test, N = 34, W = 564, P < 0.0001
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Junco preen gland bacterial communities
Highly diverse
Cluster by nest
Nestlings closely resemble each other and mother
Reliable transmission across generations via physical contact
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Acknowledgments
Photo by Nicole Gerlach
Tracy Teal
Arvind Venkataraman
Ellen Ketterson
Samuel Slowinski