Cloacal bacterial communities of tree swallows (Tachycineta bicolor): Similarity within a population, but not between pair-bonded social partners.

Host-associated microbial communities can influence the overall health of their animal hosts, and many factors, including behavior and physiology, can impact the formation of these complex communities. Bacteria within these communities can be transmitted socially between individuals via indirect (e.g., shared environments) or direct (e.g., physical contact) pathways. Limited research has been done to investigate how social interactions that occur in the context of mating shape host-associated microbial communities. To gain a better understanding of these interactions and, more specifically, to assess how mating behavior shapes an animal's microbiome, we studied the cloacal bacterial communities of a socially monogamous yet genetically polygynous songbird, the North American tree swallow (Tachycineta bicolor). We address two questions: (1) do the cloacal bacterial communities differ between female and male tree swallows within a population? and (2) do pair-bonded social partners exhibit more similar cloacal bacterial communities than expected by chance? To answer these questions, we sampled the cloacal microbiome of adults during the breeding season and then used culture-independent, 16S rRNA gene amplicon sequencing to assess bacterial communities. Overall, we found that the cloacal bacterial communities of females and males were similar, and that the communities of pair-bonded social partners were not more similar than expected by chance. Our results suggest that social monogamy does not correlate with an increased similarity in cloacal bacterial community diversity or structure. As social partners were not assessed at the same time, it is possible that breeding stage differences masked social effects on bacterial community diversity and structure. Further, given that tree swallows exhibit high variation in rates of extra-pair activity, considering extra-pair activity when assessing cloacal microbial communities may be important for understanding how these bacterial communities are shaped. Further insight into how bacterial communities are shaped will ultimately shed light on potential tradeoffs associated with alternative behavioral strategies and socially-transmitted microbes.

Cloacal microbiomes and ecology of individual barn swallows.

Microbiomes can be considered as 'second genomes' for the host, and can deeply affect its physiology, behaviour and fitness. We investigated the cloacal microbiomes (CMs) of adult and nestling barn swallows (Hirundo rustica), a small insectivorous migratory passerine bird, in order to assess whether CM structure was related to major ecological traits of individuals. Illumina sequencing of the V5-V6 hypervariable regions of the bacterial 16S rRNA gene showed that barn swallow CMs were dominated by Proteobacteria, Firmicutes, Actinobacteria, Tenericutes and Bacteroidetes. Nestling CMs were more similar to one another than adult ones, but showed higher alpha diversity. Sibling nestlings had more similar CMs than non-sibling ones. CMs of adult males also differed from those of adult females, but pair members had more similar CMs than expected by chance. In contrast, CMs did not differ between male and female nestlings. Finally, in adults, CMs strongly different from the 'average' CM were associated to lower survival prospects of the host. The CMs of a bird species in the wild are therefore related to important traits of individuals, such as survival, suggesting that microbiomes should be included among the traits examined in ecological studies.

[Intestinal Microbiota in Migrating Barn Swallows around Osaka].

Migratory birds are considered as vectors of infectious diseases, owing to their potential for transmitting pathogens over large distances. The populations of barn swallow (Hirundo rustica) migrate from Southeast Asia to the Japanese mainland during spring and migrate back to Southeast Asia during autumn. This migratory population is estimated to comprise approximately hundreds to thousands of individuals per year. However, to date, not much is known about the gastrointestinal microbiota of the barn swallow. In this study, we characterized the fecal bacterial community in barn swallow. Using 16S rRNA gene metagenomic sequencing analysis, we examined the presence and composition of potentially pathogenic bacteria in the fecal samples, which were collected during spring season from Osaka. The number (±S.D.) of total bacteria was approximately 2.1(±3.4)×108 per gram of feces. In most samples, the bacterial community composition was dominated by families, such as Enterobacteriaceae, Pseudomonadaceae, Mycoplasmataceae, Enterococcaceae, Streptococcaceae, and Alcaligenaceae. However, no relationship was found between the bacterial community composition and geographical area in the fecal samples. Potentially pathogenic bacteria were detected at the rate of >0.1%, which included Pseudomonas spp., Escherichia/Shigella spp., Enterobacter spp., Yersinia spp., Mycoplasma spp., Enterococcus spp., Achromobacter spp., and Serratia spp. Our results suggested that barn swallow is instrumental in the transmission of these genera over large distances.

Defenses against keratinolytic bacteria in birds living in radioactively contaminated areas.

Microorganisms have shaped the evolution of a variety of defense mechanisms against pathogenic infections. Radioactivity modifies bacterial communities and, therefore, bird hosts breeding in contaminated areas are expected to adapt to the new bacterial environment. We tested this hypothesis in populations of barn swallows (Hirundo rustica) from a gradient of background radiation levels at Chernobyl and uncontaminated controls from Denmark. Investment in defenses against keratinolytic bacteria was measured from feather structure (i.e., susceptibility to degradation) and uropygial secretions. We studied degradability of tail feathers from areas varying in contamination in laboratory experiments using incubation of feathers with a feather-degrading bacterium, Bacillus licheniformis, followed by measurement of the amount of keratin digested. The size of uropygial glands and secretion amounts were quantified, followed by antimicrobial tests against B. licheniformis and quantification of wear of feathers. Feathers of males, but not of females, from highly contaminated areas degraded at a lower rate than those from medium and low contamination areas. However, feathers of both sexes from the Danish populations showed little evidence of degradation. Individual barn swallows from the more contaminated areas of Ukraine produced the largest uropygial secretions with higher antimicrobial activity, although wear of feathers did not differ among males from different populations. In Denmark, swallows produced smaller quantities of uropygial secretion with lower antimicrobial activity, which was similar to swallow populations from uncontaminated areas in Ukraine. Therefore, barn swallows breeding in contaminated areas invested more in all defenses against keratinolytic bacteria than in uncontaminated areas of Ukraine and Denmark, although they had similar levels of feather wear. Strong natural selection exerted by radioactivity may have selected for individuals with higher defense capacity against bacterial infections during the 30 years since the Chernobyl disaster.

Stress response, gut microbial diversity and sexual signals correlate with social interactions.

Theory predicts that social interactions are dynamically linked to phenotype. Yet because social interactions are difficult to quantify, little is known about the precise details on how interactivity is linked to phenotype. Here, we deployed proximity loggers on North American barn swallows (Hirundo rustica erythrogaster) to examine intercorrelations among social interactions, morphology and features of the phenotype that are sensitive to the social context: stress-induced corticosterone (CORT) and gut microbial diversity. We analysed relationships at two spatial scales of interaction: (i) body contact and (ii) social interactions occurring between 0.1 and 5 m. Network analysis revealed that relationships between social interactions, morphology, CORT and gut microbial diversity varied depending on the sexes of the individuals interacting and the spatial scale of interaction proximity. We found evidence that body contact interactions were related to diversity of socially transmitted microbes and that looser social interactions were related to signalling traits and CORT.

Relationships among plumage coloration, blood selenium concentrations and immune responses of adult and nestling tree swallows.

In a number of taxa, males and females both display ornaments that may be associated with individual quality and could be reliable signals to potential mates or rivals. We examined the iridescent blue/green back and white breast of adult tree swallows (Tachycineta bicolor) to determine whether plumage reflectance is related to adult or offspring immune responses. We simultaneously addressed the influence of blood selenium levels and the interaction between blood selenium and plumage coloration on adult and nestling immunity. Selenium is a well-known antioxidant necessary for mounting a robust immune response but its importance in wild birds remains poorly understood. In females, the brightness of white breast coloration was positively associated with bactericidal capacity, but there was no association with blood selenium. In contrast, male bactericidal capacity was associated with an interactive effect between dorsal plumage coloration and blood selenium concentration. Males with bluer hues and greater blue chroma showed increased bactericidal capacity as blood selenium concentrations increased, while bactericidal capacity declined in greener males at higher blood selenium concentrations. In nestlings, bactericidal capacity was positively associated with nestling blood selenium concentrations and white brightness of both social parents. These results suggest that white plumage reflectance is indicative of quality in tree swallows and that greater attention should be paid to the reflectance of large white plumage patches. Additionally, the role of micronutrients, such as selenium, in mediating relationships between physiology and signals of quality, should be explored further.

Eggshell bacterial load is related to antimicrobial properties of feathers lining barn swallow nests.

The use of feathers to line bird's nests has traditionally been interpreted as having a thermoregulatory function. Feather-degrading bacteria growing on feathers lining nests may have antimicrobial properties, which may provide an additional benefit to lining nests with feathers. We test the hypothesis that the production of antimicrobial substances by feather bacteria affects the microbiological environment of the nest, and therefore the bacterial density on eggshells and, indirectly, hatching success. These effects would be expected to differ between nests lined with pigmented and white feathers, because bacteria grow differently on feathers of different colors. We experimentally manipulated the composition of pigmented and unpigmented feathers in nests of the barn swallow (Hirundo rustica) and studied the antimicrobial properties against the keratin-degrading bacterium Bacillus licheniformis of bacteria isolated from feathers of each color. Analyzed feathers were collected at the end of the incubation period, and antimicrobial activity was defined as the proportion of bacteria from the feathers that produce antibacterial substances effective against B. licheniformis. Our experimental manipulation affected antimicrobial activity, which was higher in nests with only white feathers at the beginning of incubation. Moreover, white feathers showed higher antimicrobial activity than black ones. Interestingly, antimicrobial activity in feathers of one of the colors correlated negatively with bacterial density on feather of the opposite color. Finally, antimicrobial activity of white feathers was negatively related to eggshell bacterial load. These results suggest that antimicrobial properties of feathers in general and of white feathers in particular affect the bacterial environment in nests. This environment in turn affects the bacterial load on eggshells, which may affect hatching success.

Cryptococcus neoformans isolation from swallow (Hirundo rustica) excreta in Iran.

Cryptococcus neoformans is an encapsulated yeast that can cause cryptococcosis, a life-threatening infection that mainly occurs in immunocompromised patients. The major environmental sources of C. neoformans have been shown to be soil contaminated with avian droppings. In the present study, we evaluated the isolation of C. neoformans from swallow (Hirundo rustica) excreta in two northern cities of Iran. Ninety-seven swallow droppings were evaluated and 498 yeast-like colonies were isolated and identified as Rhodotorula spp. (62.8%), Candida spp. (28.5%)and C. neoformans (8.7%). Cryptococcus neoformans was isolated from 5/97 (5.2%) of collected samples. Min-Max colony forming units (CFU) per one gram for the positive samples were 3-10 C. neoformans colonies. The total mean CFU per one gram for the positive samples was 4.8. The results of this study demonstrate that excreta of swallow may harbor different species of potentially pathogenic yeasts, mainly C. neoformans, and may be capable of disseminating these fungi in the environment.

Cryptococcus neoformans isolation from swallow (Hirundo rustica) excreta in Iran / Isolamento de Cryptococcus neoformans de excrementos de andorinhas (Hirundo rustica) do Irã

Cryptococcus neoformans is an encapsulated yeast that can cause cryptococcosis, a life-threatening infection that mainly occurs in immunocompromised patients. The major environmental sources of C. neoformans have been shown to be soil contaminated with avian droppings. In the present study, we evaluated the isolation of C. neoformans from swallow (Hirundo rustica) excreta in two northern cities of Iran. Ninety-seven swallow droppings were evaluated and 498 yeast-like colonies were isolated and identified as Rhodotorula spp. (62.8 percent), Candida spp. (28.5 percent)and C. neoformans (8.7 percent). Cryptococcus neoformans was isolated from 5/97 (5.2 percent) of collected samples. Min-Max colony forming units (CFU) per one gram for the positive samples were 3-10 C. neoformans colonies. The total mean CFU per one gram for the positive samples was 4.8. The results of this study demonstrate that excreta of swallow may harbor different species of potentially pathogenic yeasts, mainly C. neoformans, and may be capable of disseminating these fungi in the environment.

Cryptococcus neoformans é levedura encapsulada que pode causar criptococose, infecção potencialmente mortal que ocorre principalmente em pacientes imunocomprometidos. As principais fontes ambientais de C. neoformans são o solo contaminado com fezes de aves. No presente estudo, avaliamos o isolamento de C. neoformans de excreta de andorinhas (Hirundo rustica) em duas cidades do norte do Irã. Noventa e sete amostras de fezes de andorinhas foram avaliadas e 498 colonias semelhantes à levedura foram isoladas e identificadas como Rhodotorula spp. (62,8 por cento), Candida spp. (28,5 por cento), C. neoformans (8,7 por cento). Cryptococcus neoformans foi isolado a partir de 5/97 (5,2 por cento) das amostras coletadas. Unidades Min-Max formadoras de colonias (CFU) por 1 grama das amostras positivas foram 3-10 coloniasde C. neoformans. A média total de CFU por 1 grama das amostras positivas foi de 4,8. Os resultados deste estudo demonstram que excrementos de andorinhas podem abrigar diferentes espécies de leveduras potencialmente patógenas, principalmente C. neoformans, e podem ser capazes de disseminar estes fungos no meio ambiente.

Experimental evidence that keeping eggs dry is a mechanism for the antimicrobial effects of avian incubation.

Avian incubation dramatically reduces the abundance and diversity of microbial assemblages on eggshells, and this effect has been hypothesized as an adaptive explanation for partial incubation, the bouts of incubation that some birds perform during the egg-laying period. However, the mechanisms for these antimicrobial effects are largely unknown. In this study, we hypothesized that microbial inhibition is partly achieved through removal of liquid water, which generally enhances microbial growth, from eggshells, and experimentally tested this hypothesis in two ways. First, we placed the first- and second-laid eggs of tree swallow (Tachycineta bicolor) clutches in unincubated holding nests with either ambient or increased water on eggshells. Second, we added water to eggshells in naturally partially incubated nests. We compared microbial growth on shells during a 5-day experimental period and found that, as predicted, both unincubated groups had higher microbial growth than naturally partially incubated controls, and that only in the absence of incubation did wetted eggs have higher microbial growth than unwetted eggs. Thus, we have shown that water increases microbial growth on eggshells and that incubation nullifies these effects, suggesting that removal of water from egg surfaces is one proximate mechanism for the antimicrobial effects of incubation.

Microorganisms associated with feathers of barn swallows in radioactively contaminated areas around chernobyl.

The Chernobyl catastrophe provides a rare opportunity to study the ecological and evolutionary consequences of low-level, environmental radiation on living organisms. Despite some recent studies about negative effects of environmental radiation on macroorganisms, there is little knowledge about the effect of radioactive contamination on diversity and abundance of microorganisms. We examined abundance patterns of total cultivable bacteria and fungi and the abundance of feather-degrading bacterial subset present on feathers of barn swallows (Hirundo rustica), a colonial migratory passerine, around Chernobyl in relation to levels of ground level environmental radiation. After controlling for confounding variables, total cultivable bacterial loads were negatively correlated with environmental radioactivity, whereas abundance of fungi and feather-degrading bacteria was not significantly related to contamination levels. Abundance of both total and feather-degrading bacteria increased with barn swallow colony size, showing a potential cost of sociality. Males had lower abundance of feather-degrading bacteria than females. Our results show the detrimental effects of low-level environmental radiation on total cultivable bacterial assemblage on feathers, while the abundance of other microorganism groups living on barn swallow feathers, such as feather-degrading bacteria, are shaped by other factors like host sociality or host sex. These data lead us to conclude that the ecological effects of Chernobyl may be more general than previously assumed and may have long-term implications for host-microbe interactions and overall ecosystem functioning.

Barn swallows (Hirundo rustica) test negative for Salmonella.

Farmers around the world have been urged to eliminate barn swallows (Hirundo rustica) from their buildings, based on the unproven assumption that these birds amplify and spread Salmonella to livestock, humans, and stored food. However, we tested over 500 barn swallows in Sweden and found no evidence that they carry Salmonella. Our results cast doubt on the hypothesis that barn swallows are competent reservoirs of Salmonella and suggest that their role, if any, is limited to being accidental hosts of the bacterium.