Very high MHC Class IIB diversity without spatial differentiation in the mediterranean population of greater Flamingos.

BACKGROUND: Selective pressure from pathogens is thought to shape the allelic diversity of major histocompatibility complex (MHC) genes in vertebrates. In particular, both local adaptation to pathogens and gene flow are thought to explain a large part of the intraspecific variation observed in MHC allelic diversity. To date, however, evidence that adaptation to locally prevalent pathogens maintains MHC variation is limited to species with limited dispersal and, hence, reduced gene flow. On the one hand high gene flow can disrupt local adaptation in species with high dispersal rates, on the other hand such species are much more likely to experience spatial variation in pathogen pressure, suggesting that there may be intense pathogen mediated selection pressure operating across breeding sites in panmictic species. Such pathogen mediated selection pressure operating across breeding sites should therefore be sufficient to maintain high MHC diversity in high dispersing species in the absence of local adaptation mechanisms. We used the Greater Flamingo, Phoenicopterus roseus, a long-lived colonial bird showing a homogeneous genetic structure of neutral markers at the scale of the Mediterranean region, to test the prediction that higher MHC allelic diversity with no population structure should occur in large panmictic populations of long-distance dispersing birds than in other resident species. RESULTS: We assessed the level of allelic diversity at the MHC Class IIB exon 2 from 116 individuals born in four different breeding colonies of Greater Flamingo in the Mediterranean region. We found one of the highest allelic diversity (109 alleles, 2 loci) of any non-passerine avian species investigated so far relative to the number of individuals and loci genotyped. There was no evidence of population structure between the four major Mediterranean breeding colonies. CONCLUSION: Our results suggest that local adaptation at MHC Class IIB in Greater Flamingos is constrained by high gene flow and high MHC diversity appears to be maintained by population wide pathogen-mediated selection rather than local pathogen-mediated selection. Further understanding of how pathogens vary across space and time will be crucial to further elucidate the mechanisms maintaining MHC diversity in species with large panmictic populations and high dispersal rates.

Assessing sex-related chick provisioning in greater flamingo Phoenicopterus roseus parents using capture-recapture models.

In sexually dimorphic species, the parental effort of the smaller sex may be reduced due to competitive exclusion in the feeding areas by the larger sex or physiological constraints. However, to determine gender effects on provisioning patterns, other intrinsic and extrinsic factors affecting parental effort should be accounted for. Greater flamingos (Phoenicopterus roseus) exhibit sexual size dimorphism. In Fuente de Piedra colony, the lake dries out almost completely during the breeding season and both parents commute between breeding and foraging sites >130 km away during the chick-rearing period. Applying multistate capture-recapture models to daily observations of marked parents, we determined the effects of sex, and their interactions with other intrinsic and extrinsic factors, on the probability of chick desertion and sojourn in the colony and feeding areas. Moreover, using stable isotopes in the secretions that parents produce to feed their chicks, we evaluated sex-specific use of wetlands. The probability of chick attendance (complementary to chick desertion) was >0.98. Chick desertion was independent of parental sex, but decreased with parental age. Females stayed in the feeding areas for shorter periods [mean: 7.5 (95% CI: 6.0-9.4) days] than males [9.2 (7.3-11.8) days]. Isotopic signatures of secretions did not show sex differences in δ(13)C, but males' secretions were enriched in δ(15)N, suggesting they fed on prey of higher trophic levels than females. Both parents spent approximately 1 day in the colony, but females prolonged their mean stay when the lake dried out. Females also allocated more time to foraging in the flooded areas remaining in the colony, likely because they were energetically more stressed than males. The results indicate that sex-specific provisioning behaviour in greater flamingo is related to differential effects of both intrinsic and extrinsic factors. Males seem forage less efficiently than females, whereas females' body condition seems to be lower after feeding the chick. Our methodology may be extended to species that feed on distant food sources and that do not visit their offspring daily, to elucidate patterns of chick-provisioning behaviour.

The color of greater flamingo feathers fades when no cosmetics are applied.

Greater flamingos use cosmetic coloration by spreading uropygial secretions pigmented with carotenoids over their feathers, which makes the plumage redder. Because flamingos inhabit open environments that receive direct solar radiation during daytime, and carotenoids bleach when exposed to solar radiation, we expected that the plumage color would fade if there is no maintenance for cosmetic purposes. Here, we show that the concentrations of pigments inside feathers and on the surface of feathers were correlated, as well as that there was a correlation between the concentrations of pigments in the uropygial secretions and on the surface of feathers. There was fading in color (becoming less red) in feathers that received direct solar radiation when there was no plumage maintenance, but not so in others maintained in darkness. When we controlled for the initial color of feathers, the feathers of those individuals with higher concentration of pigments on the feather surfaces were those that lost less coloration after experimental exposure of feathers to sunny conditions. These results indicate that exposure to sunlight is correlated with the fading of feather color, which suggests that individuals need to regularly apply makeup to be more colorful. These results also reinforce the view that these birds use cosmetic coloration as a signal amplifier of plumage color. This may be important in species using highly variable habitats, such as wetlands, since the conditions experienced when molting may differ from those when the signal should be functional, usually months after molting.

Bioaccumulation of trace elements affects chick body condition and gut microbiome in greater flamingos.

Elevated concentrations of trace elements represent a major concern to wetland ecosystems, since river estuaries are geochemical endpoints that accumulate pollution. Although the negative impact of environmental exposure of highly toxic elements such as Pb and Hg has received substantial attention, we still lack a comprehensive understanding of the effects that these and other common trace elements have on natural populations. We used greater flamingos as a study system within three sites that represent a gradient of pollution. Controlling for environmental sediment exposure, we assessed if signatures of bioaccumulation in feathers for ten trace elements (As, Cd, Cr, Cu, Hg, Ni, Pb, Se, Sn and Zn) are associated with two known proxies of health: body condition and the gut bacterial microbiome. We found evidence of an adverse effect of Se, Hg, and Pb bioaccumulation on body condition. Furthermore, bioaccumulation of the elements As, Cu, Se, Pb and Zn influenced different aspects of the gut microbiome. Bioaccumulation of Se led to a shift in the microbiome composition, largely driven by an enrichment of Bacteroides plebeius, which is linked to the breakdown of sulphated polysaccharides of algae. Bacteroides plebeius was negatively associated with chick body condition, suggesting an adverse effect of a microalgae diet rich in Se. Pb bioaccumulation was linked with a decrease in microbial diversity (adjusted-R2 = 10.4%) and an increase in heterogeneity of the microbial community (adjusted-R2 = 10.5%), an indication of impaired gut homeostasis. As, Cu and Zn had more nuanced effects on gut microbiome heterogeneity according to breeding site and bioaccumulation concentration. Our results therefore suggest that in addition to well-studied elements, bioaccumulation of poorly studied elements also adversely affect health of natural populations.

Experience-dependent natal philopatry of breeding greater flamingos.

1. Contrary to the generally high level of natal philopatry (i.e. likelihood that individuals breed at their natal colony) found in first-breeding colonial birds, little is known of natal philopatry later in life. Most hypotheses advanced to explain natal philopatry are valid at all ages. However, for young and inexperienced birds, the benefits of natal philopatry may be counterbalanced by the costs of intraspecific competition at the natal colony making dispersal temporarily advantageous. In turn, experience may increase competitive ability and make natal philopatry advantageous again. 2. We evaluated this hypothesis on the large-scale dispersal of greater flamingos Phoenicopterus roseus breeding among three colonies comprising >85% of the Western Mediterranean metapopulation. The Camargue (France) and Fuente de Piedra (Spain) are large and saturated colonies while Molentargius (Sardinia) is a recent and growing colony. 3. We used a 20-year capture-mark-resighting dataset of 4900 flamingos ringed as chicks in Camargue and Fuente de Piedra and breeding at the three colonies. We assessed the effects of natal colony and breeding experience (first-time observed breeders versus confirmed experienced breeders) on dispersal using multistate capture-recapture models. Dispersal to an unobservable state accounted for temporary emigration. 4. Fidelity was higher at the natal colony (>84%) than elsewhere. Fidelity increased with experience in the two large colonies (Camargue and Fuente de Piedra) suggesting a large-scale experience-related despotic distribution. Breeding dispersal was significant (up to 61% and 52% for first-time breeders and experienced breeders, respectively) so that colony dynamics is affected by exchanges with other colonies. Except for Fuente-born breeders leaving Molentargius, dispersal to the natal colony was higher than to any other colonies. 5. Survival was not higher at the natal colony. Inexperienced birds likely had lower breeding success at the Camargue and skipped reproduction after having emigrated to the other large colony but not to Molentargius. Breeding at Molentargius could allow avoiding queuing (and non-breeding) at the large colonies while gaining experience and competitive ability for future attempts. 6 Natal philopatry appears as an important driver of large-scale breeding dispersal in the Greater flamingo. The fitness advantage of natal philopatry is likely experience-dependent and mediated by the variations of intraspecific competition.

Offspring Microbiomes Differ Across Breeding Sites in a Panmictic Species.

High dispersal rates are known to homogenize host's population genetic structure in panmictic species and to disrupt host local adaptation to the environment. Long-distance dispersal might also spread micro-organisms across large geographical areas. However, so far, to which extent selection mechanisms that shape host's population genetics are mirrored in the population structure of the enteric microbiome remains unclear. High dispersal rates and horizontal parental transfer may homogenize bacterial communities between breeding sites (homogeneous hypothesis). Alternatively, strong selection from the local environment may differentiate bacterial communities between breeding sites (heterogeneous hypothesis). Furthermore, selection from age-specific environmental or physiological factors may differentiate the microbiome between juveniles and adults. Here, we analyzed the cloacal bacterial 16S rRNA gene of fledgling greater flamingos, Phoenicopterus roseus, across nine western Mediterranean breeding sites and four breeding seasons (n = 731) and adult birds (n = 27) from a single site. We found that fledgling cloacal microbiome, as measured by alpha diversity, beta diversity, the relative abundance of assigned sequence variants (ASVs) belonging to a phylum and genus composition within phylum, varied significantly between sampling sites and across time within site despite high adult dispersal rates. The spatio-temporal effects were stronger on individual ASV absence/presence than on ASV abundance (i.e., than on core microbiome composition). Spatial effects had a stronger effect than temporal effects, particularly on ASV abundance. Our study supports the heterogeneous hypothesis whereby local environmental conditions select and differentiate bacterial communities, thus countering the homogenizing effects of high-dispersing host species. In addition, differences in core microbiome between adult vs. fledgling samples suggests that differences in age-specific environmental and/or physiological factors result in differential selection pressure of core enteric microbiome between age classes, even within the same environment. In particular, the genus Corynebacterium, associated with both seasonal fat uptake and migration in previous studies, was much more abundant in high-dispersing fledglings than in more resident adults. To conclude, selection mechanisms that shape the host's genetic structure cannot be extended to the genetic structure of the enteric microbiome, which has important implications regarding our understanding of both host local adaptation mechanisms and enteric microbiome population genetics.

Flamingos and drought as drivers of nutrients and microbial dynamics in a saline lake.

Waterbird aggregations and droughts affect nutrient and microbial dynamics in wetlands. We analysed the effects of high densities of flamingos on nutrients and microbial dynamics in a saline lake during a wet and a dry hydrological year, and explored the effects of guano on prokaryotic growth. Concentrations of dissolved organic carbon, total phosphorus and total nitrogen in the surface waters were 2-3 fold higher during the drought and were correlated with salinity. Flamingos stimulated prokaryotic heterotrophic production and triggered cascading effects on prokaryotic abundance, viruses and dissolved nitrogen. This stimulus of heterotrophic prokaryotes was associated with soluble phosphorus inputs from guano, and also from sediments. In the experiments, the specific growth rate and the carrying capacity were almost twice as high after guano addition than in the control treatments, and were coupled with soluble phosphorus assimilation. Flamingo guano was also rich in nitrogen. Dissolved N in lake water lagged behind the abundance of flamingos, but the causes of this lag are unclear. This study demonstrates that intense droughts could lead to increases in total nutrients in wetlands; however, microbial activity is likely constrained by the availability of soluble phosphorus, which appears to be more dependent on the abundance of waterbirds.

Temporal and spatial variation in dispersal in the greater flamingo (Phoenicopterus ruber roseus).

We studied movements of individually marked greater flamingos (Phoenicopterus ruber roseus) born in the Camargue, southern France, between their two most important breeding colonies in the western Mediterranean (Camargue and Fuente de Piedra, Spain) from 1986 to 1992. The two sites differ in the frequency with which they offer suitable conditions for breeding. Flamingos have bred each year in the Camargue since 1974, but in only 12 of the past 22 years at Fuente de Piedra. Higher colony fidelity is thus expected in the less variable environment (Camargue), but if dispersal occurs competition might be an important factor causing this dispersal. Following years during which breeding birds in the Camrgue were disturbed (1988 and 1990) a higher proportion of adults changed colonies between breeding attempts (= breeding dispersal, 12.4%), while only 0.4% of flamingos breeding in the Camargue dispersed in the other years. As expected, flamingos breeding at Fuente de Piedra showed a higher rate of breeding dispersal (8.14%). No differences were observed between males and females. The importance of breeding failure as a factor causing breeding dispersal in flamingos was also confirmed by the movements of individual birds. The proportion of young flamingos that moved from their natal colony to start breeding at Fuente de Piedra (= natal dispersal) was independent of sex and age, but increased when breeding access to the Camargue colony was more difficult. However, natal dispersal was also higher in 1988 and 1990 (40.5%) than in the remaining years (1.2%), as was breeding dispersal. We discuss possible ways in which the increased natal dispersal among inexperienced birds could be linked with the increased breeding dispersal of adults in the same year.

Interannual variations in feeding frequencies and food quality of greater flamingo chicks (Phoenicopterus roseus): evidence from plasma chemistry and effects on body condition.

Greater flamingos in southern Spain foraged in areas distant from a breeding site, spending 4-6 days in foraging areas between successive visits to the colony to feed their chicks. During four years, we took blood samples from chicks to ascertain whether there were interannual variations in several blood parameters, indicative of food quality and feeding frequencies. When the chicks were captured, 20-31% of them had their crops empty, indicating that not all chicks were fed daily. Additional evidence of variations in feeding frequencies was obtained from a principal component analysis (PCA) on plasma chemistry values, which also indicated that there were annual variations in the quality of food received by chicks. The association of cholesterol and glucose with some PC axes indicated that some chicks were experiencing fasting periods. Of all plasma metabolites considered, cholesterol was the best one to predict body condition. Greater flamingo chicks experiencing longer fasting intervals, as suggested by higher plasma levels of cholesterol, were in lower body condition.