Season as a discriminating factor for faecal metabolomic composition of great tits (Parus major)

The microbiome of wild birds has been associated with health status and risk of disease development, but underlying metabolomic mechanisms are still unknown. Metabolites produced by microbial organisms may affect host metabolic processes and by doing so influence host health. Here we provide for the first time data on the faecal metabolome of wild great tits (Parus major) by analyzing metabolites associations with age, sex, season and body condition. Using untargeted metabolomics, we analyzed faecal samples from 112 great tits that were caught in a deciduous forest fragment in Flanders (Belgium) during late autumn and 19 animals that were re-captured during early spring. In this study, no significant associations between the faecal metabolites and age, sex and body condition were observed. However, season was shown to be a discriminating factor for the metabolomic composition of great tits, suggesting an impact of environmental factors

Mitigating the impact of microbial pressure on great (Parus major) and blue (Cyanistes caeruleus) tit hatching success through maternal immune investment

he hatching success of a bird’s egg is one of the key determinants of avian reproductive success, which may be compromised by microbial infections causing embryonic death. During incubation, outer eggshell bacterial communities pose a constant threat of pathogen translocation and embryo infection. One of the parental strategies to mitigate this threat is the incorporation of maternal immune factors into the egg albumen and yolk. It has been suggested that habitat changes like forest fragmentation can affect environmental factors and life-history traits that are linked to egg contamination. This study aims at investigating relationships between microbial pressure, immune investment and hatching success in two abundant forest bird species and analyzing to what extent these are driven by extrinsic (environmental) factors. We here compared (1) the bacterial load and composition on eggshells, (2) the level of immune defenses in eggs, and (3) the reproductive success between great (Parus major) and blue (Cyanistes caeruleus) tits in Belgium and examined if forest fragmentation affects these parameters. Analysis of 70 great tit and 34 blue tit eggshells revealed a similar microbiota composition (Enterobacteriaceae, Lactobacillus spp., Firmicutes and Bacteroidetes), but higher bacterial loads in great tits. Forest fragmentation was not identified as an important explanatory variable. Although a significant negative correlation between hatching success and bacterial load on the eggshells in great tits corroborates microbial pressure to be a driver of embryonic mortality, the overall hatching success was only marginally lower than in blue tits. This may be explained by the significantly higher levels of lysozyme and IgY in the eggs of great tits, protecting the embryo from increased infection pressure. Our results show that immune investment in eggs is suggested to be a species-specific adaptive trait that serves to protect hatchlings from pathogen pressure, which is not directly linked to habitat fragmentation

Salmonella Typhimurium DT193 and DT99 are present in great and blue tits in Flanders, Belgium

Endemic infections with the common avian pathogen Salmonella enterica subspecies enterica serovar Typhimurium (Salmonella Typhimurium) may incur a significant cost on the host population. In this study, we determined the potential of endemic Salmonella infections to reduce the reproductive success of blue (Cyanistes caeruleus) and great (Parus major) tits by correlating eggshell infection with reproductive parameters. The fifth egg of each clutch was collected from nest boxes in 19 deciduous forest fragments. Out of the 101 sampled eggs, 7 Salmonella Typhimurium isolates were recovered. The low bacterial prevalence was reflected by a similarly low serological prevalence in the fledglings. In this study with a relatively small sample size, presence of Salmonella did not affect reproductive parameters (egg volume, clutch size, number of nestlings and number of fledglings), nor the health status of the fledglings. However, in order to clarify the impact on health and reproduction a larger number of samples have to be analyzed. Phage typing showed that the isolates belonged to the definitive phage types (DT) 193 and 99, and multi-locus variable number tandem repeat analysis (MLVA) demonstrated a high similarity among the tit isolates, but distinction to human isolates. These findings suggest the presence of passerine-adapted Salmonella strains in free-ranging tit populations with host pathogen co-existence

Investigation of amphibian mortality events in wildlife reveals an on-going ranavirus epidemic in the North of the Netherlands

In the four years following the first detection of ranavirus (genus Ranavirus, family Iridoviridae) infection in Dutch wildlife in 2010, amphibian mortality events were investigated nationwide to detect, characterize and map ranaviruses in amphibians over time, and to establish the affected host species and the clinico-pathological presentation of the disease in these hosts. The ultimate goal was to obtain more insight into ranavirus disease emergence and ecological risk. In total 155 dead amphibians from 52 sites were submitted between 2011 and 2014, and examined using histopathology, immunohistochemistry, virus isolation and molecular genetic characterization. Ranavirus-associated amphibian mortality events occurred at 18 sites (35%), initially only in proximity of the 2010 index site. Specimens belonging to approximately half of the native amphibian species were infected, including the threatened Pelobates fuscus (spadefoot toad). Clustered massive outbreaks involving dead adult specimens and ranavirus genomic identity indicated that one common midwife toad virus (CMTV)-like ranavirus strain is emerging in provinces in the north of the Netherlands. Modelling based on the spatiotemporal pattern of spread showed a high probability that this emerging virus will continue to be detected at new sites (the discrete reproductive power of this outbreak is 0.35). Phylogenetically distinct CMTV-like ranaviruses were found in the south of the Netherlands more recently. In addition to showing that CMTV-like ranaviruses threaten wild amphibian populations not only in Spain but also in the Netherlands, the current spread and risk of establishment reiterate that understanding the underlying causes of CMTV-like ranavirus emergence requires international attention

Endemism of Salmonella Typhimurium does not compromise reproductive success in great (Parus major) and blue (Cyanistes caeruleus) tits

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Usutu virus epizootic and Plasmodium coinfection in Eurasian blackbirds (Turdus merula) in Flanders, Belgium

At the end of the summer of 2016, unusually high levels of mortality were detected in Passeriformes and Strigiformes in Flanders, Belgium, mainly in Eurasian Blackbirds (Turdus merula). A passive surveillance program demonstrated a widespread Usutu virus outbreak and revealed a coinfection with Plasmodium in 99% of the dead passerine birds that were necropsied

Exploring the faecal microbiome of the Eurasian nuthatch (Sitta europaea)

Gastrointestinal microbiota fulfill pivotal roles in providing a host with nutrition and protection from pathogenic microorganisms. Up to date, most microbiota research has focused on humans and other mammals, whereas birds and especially wild birds lag behind. Within the field of the avian gut microbiome, research is heavily biased towards poultry. In this study, we analyzed the gut microbiome of the Eurasian nuthatch (Sitta europaea), using faecal samples of eight nestlings originating from three nuthatch nests in the south of Ghent (Belgium), using Illumina sequencing of the 16S rRNA gene. Relative frequency analysis showed a dominance of Firmicutes and Actinobacteria and to a lesser extent Proteobacteria. Bacteroidetes and other phyla were relatively rare. At higher taxonomic levels, a high degree of inter-individual variation in terms of overall microbiota community structure as well as dominance of certain bacteria was observed, but with a higher similarity for the nestlings sharing the same nest. When comparing the nuthatch faecal microbiome to that of great tit nestlings that were sampled during the same breeding season and in the same forest fragment, differences in the microbial community structure were observed, revealing distinct dissimilarities in the relative abundancy of taxa between the two bird species. This study is the first report on the nuthatch microbiome and serves as a reference study for nuthatch bacterial diversity and can be used for targeted screening of the composition and general functions of the avian gut microbiome

Tree species diversity and forest edge density jointly shape the gut microbiota composition in juvenile great tits (Parus major)

Despite the microbiome’s key role in health and fitness, little is known about the environmental factors shaping the gut microbiome of wild birds. With habitat fragmentation being recognised as a major threat to biological diversity, we here determined how forest structure influences the bacterial species richness and diversity of wild great tit nestlings (Parus major). Using an Illumina metabarcoding approach which amplifies the 16S bacterial ribosomal RNA gene, we measured gut microbiota diversity and composition from 49 great tit nestlings, originating from 23 different nests that were located in 22 different study plots across a gradient of forest fragmentation and tree species diversity. Per nest, an average microbiome was determined on which the influence of tree species (composition and richness) and forest fragmentation (fragment area and edge density) was examined and whether this was linked to host characteristics (body condition and fledging success). We found an interaction effect of edge density with tree species richness or composition on both the microbial richness (alpha diversity: Chao1 and Shannon) and community structure (beta diversity: weighted and unweighted UniFrac). No significant short-term impact was observed of the overall faecal microbiome on host characteristics, but rather an adverse effect of specific bacterial genera on fledging success. These results highlight the influence of environmental factors on the microbial richness as well as the phylogenetic diversity during a life stage where the birds’ microbiota is shaped, which could lead to long-term consequences for host fitness

Forest fragmentation and tree species composition jointly shape breeding performance of two avian insectivores

Habitat fragmentation and forestry practices affect forest structure and composition, and hence, their intrinsic value for biodiversity conservation. While higher tree species diversity is commonly proposed to result in habitat of higher quality for forest species, how these tree diversity and tree composition effects interact with forest fragmentation in terms of critical resources and demographic effects on forest birds remains poorly understood. We investigated here possible synergistic effects of forest fragmentation and tree species composition on the breeding performance of two common, insectivorous forest birds in a human-dominated landscape in northern Belgium. We monitored the breeding performance of Great Tits and Blue Tits in 53 plots across independent gradients of tree species composition and forest fragmentation. In addition, data on the biomass of the main food source of these two species (i.e. caterpillars) was collected during the breeding season. Both tree composition and habitat fragmentation impacted the breeding performance of Great and Blue Tits. Effects of tree species composition were mainly driven by tree species identity, rather than by tree species diversity, and the highest breeding performance was obtained in monocultures of Pedunculate Oaks. Fragmentation effects were only observed in resource-poor Beech monocultures with breeding performance declining with reduction in forest area. Structural Equation Modelling revealed diverse and species-specific pathways: for Great Tits tree composition effects on breeding performance were driven by resource availability while for Blue Tits these effects were driven by variation in clutch size. Thus, forestry practices aiming at promoting forest-dependent birds could benefit from including tree species that support high arthropod numbers and by maintaining forest patches of larger sizes