Avian haemosporidians in the cattle egret (Bubulcus ibis) from central-western and southern Africa: High diversity and prevalence.

We described the geographic distribution of 82 haemosporidian lineages (Plasmodium, Haemoproteus, and Leucocytozoon) in the cattle egret sampled in five countries in central-western and southern Africa. Seventy-three lineages have not previously been reported. We determined the prevalence of three haemosporidians in the samples. We investigated the influence of the internal environment of the host and environmental variables on the Plasmodium diversity and whether environmental variables may explain spatial variations in the prevalence of Plasmodium. We screened DNA from 509 blood samples from nestlings in 15 African colonies for infection by sequencing the cytochrome b gene of parasites. The molecular phylogenetic analysis was performed using Bayesian methods and including sequences from the MalAvi and GeneBank databases. We found 62 new Plasmodium lineages in a clade with MYCAME02, which is a lineage described in waterbirds and recently identified in birds of prey as Plasmodium paranucleophilum. Two Haemoproteus lineages identified in cattle egret formed a distinct group with Haemoproteus catharti and MYCAMH1 (Haemoproteus spp.). Seven Leucocytozoon lineages found in the cattle egret clustered with Leucocytozoon californicus. We found different Plasmodium diversities among the colonies sampled, demonstrating that the internal environment of the host is not the primary determinant of diversity. A linear mixed-effects multivariate model showed that precipitation was positively associated with Plasmodium diversity when controlling for the effects of temperature, colony composition (mixed and non-mixed species) and country. Moreover, a generalized mixed model showed that temperature was positively associated with the prevalence of Plasmodium when controlling for precipitation, elevation and country. We conclude that the cattle egret is a good model for future haemosporidian studies, as we found a significant number of new lineages in this host, which occupies regions with different climate characteristics where environmental variables exert an influence on the diversity and prevalence of Plasmodium.

Considerations for monitoring population trends of colonial waterbirds using the effective number of breeders and census estimates.

Detecting trends in population size fluctuations is a major focus in ecology, evolution, and conservation biology. Populations of colonial waterbirds have been monitored using demographic approaches to determine annual census size (Na). We propose the addition of genetic estimates of the effective number of breeders (Nb) as indirect measures of the risk of loss of genetic diversity to improve the evaluation of demographics and increase the accuracy of trend estimates in breeding colonies. Here, we investigated which methods of the estimation of Nb are more precise under conditions of moderate genetic diversity, limited sample sizes and few microsatellite loci, as often occurs with natural populations. We used the wood stork as a model species and we offered a workflow that researchers can follow for monitoring bird breeding colonies. Our approach started with simulations using five estimators of Nb and the theoretical results were validated with empirical data collected from breeding colonies settled in the Brazilian Pantanal wetland. In parallel, we estimated census size using a corrected method based on counting active nests. Both in simulations and in natural populations, the approximate Bayesian computation (ABC) and sibship assignment (SA) methods yielded more precise estimates than the linkage disequilibrium, heterozygosity excess, and molecular coancestry methods. In particular, the ABC method performed best with few loci and small sample sizes, while the other estimators required larger sample sizes and at least 13 loci to not underestimate Nb. Moreover, according to our Nb/Na estimates (values were often ≤0.1), the wood stork colonies evaluated could be facing the loss of genetic diversity. We demonstrate that the combination of genetic and census estimates is a useful approach for monitoring natural breeding bird populations. This methodology has been recommended for populations of rare species or with a known history of population decline to support conservation efforts.

Inferred kinship patterns reveal low levels of extra-pair paternity in the endangered Neotropical Jabiru Stork (Jabiru mycteria, Aves: Ciconiiformes).

The present study inferred the genetic mating system in a natural breeding population of the Jabiru Stork (Jabiru mycteria), a Neotropical wading bird considered endangered in part of its distribution range. Based on data from eight microsatellite loci, maximum-likelihood kinship reconstruction techniques, parentage assignment analyses and effective population size (Ne) estimates were applied to samples collected in the Brazilian Pantanal wetland (N = 45 nestlings from 20 nests; N = 17 shed adult feathers from 11 nests). The relationship diagnosis was determined for most of the complete clutches (86.66 %): 92.31 % were full siblings and 7.69 % were half siblings. Shed feathers collected from the nests matched the genetic parents of the offspring in 80 % of cases. Feathers collected from the ground below the nests were compatible with the putative parents in 41.67 % of cases. A mean Ne of 35 reproductive individuals was inferred, corresponding to an Ne/Nc ratio of 0.09, which is similar to the ratio found in populations of a number of different wild animals. The higher proportion of full siblings identified in the broods suggests that genetic monogamy is the prevalent mating system in the Jabiru Stork, while the detection of half siblings suggests some degree of extra-pair paternity. The present findings are in agreement with previous ecological observations of social monogamy in this species, despite the isolated evidence of extra-pair copulation events. This study also demonstrates the usefulness of a noninvasive approach to sampling adults and performing parentage and relatedness analyses in an elusive, threatened species.

Reconstructing genetic mating systems in the absence of parental information in colonially breeding waterbirds.

BACKGROUND: DNA-based studies have demonstrated that avian genetic mating systems vary widely, with many species deviating from long-assumed monogamy by practicing extra-pair paternity and conspecific brood parasitism. Colonially breeding waterbirds provide interesting models in which to investigate this question because they show nesting habits proposed to promote alternative reproductive strategies. However, little is known about the genetic mating systems of this group of birds, mainly due to difficulties in obtaining genetic data from incubating adults at nests that are necessary for conducting conventional parentage studies. Here, we inferred kinship patterns among offspring in broods of three co-distributed waterbird species, Wood Stork (Mycteria americana), Roseate Spoonbill (Platalea ajaja) and Great Egret (Ardea alba egretta), to investigate genetic mating system in the absence of parental data. RESULTS: Multi-step analyses combining estimates of relatedness coefficients, formulation of relationship-hypotheses, significance testing of alternative hypotheses, and maximum-likelihood sibship reconstruction techniques revealed evidence that alternative reproductive strategies may be present in natural populations of Wood Storks and Roseate Spoonbills, whereas relatedness of co-nestlings diagnosed in the Great Egrets did not deviate from a hypothesis of genetic monogamy. Specifically, under this analytical framework, inferred kinship relationships revealed that Great Egret nests contained full-sibling nestlings (100%), with the Roseate Spoonbill (RS) and Wood Stork (WS) exhibiting proportions of half-siblings (RS: 5%) and/or unrelated nestlings (RS: 24%; WS: 70%), patterns consistent with extra-pair paternity and conspecific brood parasitism, respectively. CONCLUSIONS: We provide evidence that genetic monogamy occurs in Brazilian natural breeding colonies of the Great Egret, but is not the sole reproductive strategy employed by the Wood Stork and the Roseate Spoonbill. In fact, extra-pair paternity and conspecific brood parasitism were common in the latter two species, with a combined frequency of 7.5% and 11.3% in Roseate Spoonbill and Wood Stork, respectively. Although geographically co-distributed, differences among these species may be due to variation in their life histories. From a methodological standpoint, the approach implemented here, although not free from limitations, can have broad application for analyzing systems with limited genealogical information and/or in studying similarly challenging organisms in which obtaining genetic data on complete families is problematic.

Genetic variability and level of differentiation among Brazilian pantanal wood stork populations.

The Wood Stork (Mycteria americana) is a colonial wading bird of the tropical and lower subtropical zones. We assessed genetic structure within and among five stork colonies from the Brazilian Pantanal and compared our data with those from North American populations. Samples of 234 individuals were studied using protein electrophoresis to evaluate genetic variability and interpopulation differentiation. Of 22 loci examined, 7 were polymorphic (mean heterozygosity = 0.068). The low Fst value (0.005) indicated little intraspecific variation among breeding colonies. Estimated number of migrants per generation based on private alleles (Nm = 11.3) and on Fst (48.8) suggests high gene flow Nei's genetic distance values among Pantanal colonies ranged from 0.0001 to 0.0034, demonstrating low genetic divergence among populations. Our data can be explained by supposing high gene flow levels among Pantanal colonies, and between North and South American populations, intermediated by a probable interbreeding population in Central America.