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Exploring the associations between preen oil bacterial, chemical and proteomic profiles of passerines.

Preen gland bacteria are thought to be the key producers of preen oil components such as chemosignalling molecules including volatile organic compounds (VOCs) and antimicrobial compounds including peptides and antimicrobial VOCs. However, data on the preen oil bacteriome and chemical composition are limited to a small subset of bird species, and the presence of antimicrobial peptides is largely unexplored. Here, we performed an exploratory study to characterize, for the first time, the preen oil chemical and proteomic profiles and to explore the possible contribution of the bacteriome to the production of preen oil VOCs and antimicrobial peptides (bacteriocins) in eight passerine species, each represented by a single individual. Preen oil bacteriome, chemical and proteomic profiles varied among birds. The bacterial profiles were dominated by the genera Streptococcus, Lactococcus, Corynebacterium and Cutibacterium. The chemical profiles mainly consisted of alcohols, ketones and carboxylic acids. The biological functions primarily associated with the proteomic profiles were proteolysis and response to oxidative stress. Although we were unable to explore a direct association between the bacteriome and chemical profiles, the preen oil contained bacteriocin- and VOC-producing bacterial genera capable of producing detected microbially-derived VOCs (mVOCs), the relative abundance of which varied between birds. Riparian species showed the highest chemical diversity and high abundances of putative preen oil mVOC-producing bacteria, which could suggest habitat-specific adaptations. This exploratory study may significantly contribute to the formulation of hypotheses on the potential role of host ecological factors in the variation of preen oil bacterial, chemical and proteomic profiles in passerines.

Animals

Coronavirus surveillance in passerines reveals novel deltacoronaviruses in Eurasian tree sparrows with implications for One Health and livestock biosecurity.

Coronaviruses (CoVs) are widespread RNA viruses infecting a broad range of avian and mammalian hosts. Although gammacoronaviruses and deltacoronaviruses (DCoVs) are common in wild birds, their presence in Eurasian passerines remains poorly understood. We screened 243 birds (35 species) at migratory stopover sites in Slovenia (2020-2021) using pan-coronavirus RT-PCR. Coronavirus RNA was detected only in four Eurasian Tree Sparrows (Passer montanus). Whole-genome sequencing yielded genomes of 26,017-26,018 bp with high internal conservation (99.95-99.98% identity). Phylogenetic analysis revealed notable evolutionary incongruence: isolates were highly related to porcine DCoVs in the ORF1ab region (95.6-96.1% amino acid identity) but clustered with divergent avian DCoVs in the spike gene (75.7-76.8% identity). RDP5 analysis provided strong evidence for a large-scale recombination event (p = 1.17 × 10-43), consistent with a mosaic genomic architecture combining an ORF1ab region closely related to porcine DCoVs with an avian-associated spike gene. This genomic pattern highlights evolutionary connectivity among DCoVs associated with different host groups and the potential role of recombination in changes in host association. The synanthropic behaviour and mobility of P. montanus facilitate contact with diverse hosts, making this species relevant for investigating DCoV ecology at wildlife-livestock interfaces. These findings represent the first genomic characterisation of DCoVs in P. montanus in Europe and support the inclusion of passerines in broader coronavirus surveillance. Genomic surveillance of underrepresented wild-bird hosts can improve our understanding of DCoV diversity, recombination, and evolution across wildlife-livestock interfaces.

Cross-species transmission

Two-Step Loss of GLUTs in the High-Metabolism Passerines.

Glucose transporters (GLUTs) play vital roles in cellular metabolism. Understanding their evolutionary dynamics in birds is essential for elucidating avian physiology and adaptation. However, the choice of gene detection method in gene family analysis may affect the conclusion. Here, we present a comprehensive investigation of methodologies and GLUT gene loss events in avian lineages, focusing on the loss of GLUT4 and GLUT8. To illustrate the effects of these methods, we first employed BUSCO-based homolog identification, calculated pairwise evolutionary distances between different species, and performed separate blastn and blastp searches to identify homologs in two groups of animals. Our analyses revealed a significant decline in blastn accuracy with increasing evolutionary distance, represented by relative divergence times. Through a more robust blastp-based gene detection pipeline, we provide evidence for the loss of GLUT genes in birds based on 58 vertebrate genomes, including 47 bird species. Our results support the reported early loss of GLUT4 in Aves. We also newly emphasize the absence of GLUT8 in passerines, potentially due to adaptation to high-sugar diets in their ancestors. These findings enhance our knowledge of avian metabolism and the evolution of GLUT genes.

Animals

Mercury residues in tissues of dead and surviving birds fed methylmercury.

Concentrations of mercury in passerine birds fed diets containing 40 ppm methylmercury were similar in tissues of birds that died from mercury poisoning and in those that were sacrificed after half the group had died. Residues were higher in tissues of birds that died, but the differences were not statistically significant. Residue levels were highest in livers, followed by kidneys and brains. Levels of mercury were similar in breast muscle, carcass, and whole body. Mercury levels were highest in redwinged blackbirds, lowest in grackles, and intermediate in starlings and cowbirds. Mercury concentrations exceeded 20 ppm in all tissues of all species and were similar to levels reported in wild birds known to have died of mercury poisoning.

Animals

Paralog-aware assembly and filtering strategies reveal minimal nucleotide variation on the macro germline-restricted chromosome of the zebra finch.

The germline-restricted chromosome (GRC) of passerines is a remarkable tissue-specific chromosome that accumulated paralogs of genes from the regular "A chromosomes" over millions of years, often amplified into dozens of gene copies. In addition to its repetitive content, typically uniparental inheritance, and lack of recombination, the GRC resembles non-recombining sex chromosomes and some B chromosomes, for all of which assembly and single-nucleotide polymorphisms (SNPs) calling are difficult. Here, we first show that much of the Australian zebra finch macro-GRC can be assembled using accurate long reads. We then describe a paralog-aware Snakemake pipeline, ParaVar, to map short reads from the GRC to retrieve GRC regions suitable for haplotype-based analysis. ParaVar reliably calls hundreds of SNPs across the GRC, thereby providing an estimate of nucleotide diversity on the highly repetitive zebra finch macro-GRC. Our results show significantly lower nucleotide diversity (20- to 50-fold lower) on the GRC compared to the mitogenome and autosomes, and a strong phylogenetic discordance between the GRC and the mitochondrial genome. Beyond the contribution of background selection, our results suggest that a single GRC haplotype recently spread through the populations while jumping across matrilines via occasional paternal inheritance. We anticipate that our paralog-aware pipeline will be useful for SNP calling and population genetics analyses of repetitive GRCs, sex chromosomes, and B chromosomes.

Animals

Stereometric observations on the hypothalamic neurosecretory system of the spotted owlet, Athene brama Temminck, in total preparations.

The configuration of the HNS of the spotted owlet, Athene brama Temminck, was studied by employing the bulk-staining technique. This enabled a three-dimensional view of the HNS. The neurons of the SON are distributed into three divisions, namely, the preoptic, median and lateral. The PVN is U-shaped; the distal portion of the arms of the U extends laterally to form the lateral division of the PVN, which remains in continuity with the lateral division of the SON. The periventricular division of the PVN joins ventrally the median division of the SON. Unlike in the passerine birds, in the spotted owlet the PVN is more prominently developed than the SON. Since the axonal pathways are not stained, the proximal portion of the hypothalamo-hypophysial tract could not be demonstrated in bulk-stained preparations. Hence it was not possible to trace with certainty the origin of the axonal terminations in the ME and in the NL. Anterior of the ME, the tract branches into two; one branch enters the zona externa to the ME and the other is continued into the NL. The ME is divisible into an AF-positive anterior region and an AF-negative posterior region. The NL is saccular and heavily loaded with the NSM. Studies on the HNS of the spotted owlet by the bulk-staining technique reveal that the general configuration of the system is comparable to that of the HNS of birds studied previously.

Animals

Repeated evolution on oceanic islands: comparative genomics reveals species-specific processes in birds.

Understanding the interplay between genetic drift, natural selection, gene flow, and demographic history in driving phenotypic and genomic differentiation of insular populations can help us gain insight into the speciation process. Comparing patterns across different insular taxa subjected to similar selective pressures upon colonizing oceanic islands provides the opportunity to study repeated evolution and identify shared patterns in their genomic landscapes of differentiation. We selected four species of passerine birds (Common Chaffinch Fringilla coelebs/canariensis, Red-billed Chough Pyrrhocorax pyrrhocorax, House Finch  Haemorhous mexicanus and Dark-eyed/island Junco Junco hyemalis/insularis) that have both mainland and insular populations. Changes in body size between island and mainland populations were consistent with the island rule. For each species, we sequenced whole genomes from mainland and insular individuals to infer their demographic history, characterize their genomic differentiation, and identify the factors shaping them. We estimated the relative (Fst) and absolute (dxy) differentiation, nucleotide diversity (π), Tajima's D, gene density and recombination rate. We also searched for selective sweeps and chromosomal inversions along the genome. All species shared a marked reduction in effective population size (Ne) upon island colonization. We found diverse patterns of differentiated genomic regions relative to the genome average in all four species, suggesting the role of selection in island-mainland differentiation, yet the lack of congruence in the location of these regions indicates that each species evolved differently in insular environments. Our results suggest that the genomic mechanisms involved in the divergence upon island colonization-such as chromosomal inversions, and historical factors like recurrent selection-differ in each species, despite the highly conserved structure of avian genomes and the similar selective factors involved. These differences are likely influenced by factors such as genetic drift, the polygenic nature of fitness traits and the action of case-specific selective pressures.

Animals

Host feeding patterns of Connecticut mosquitoes (Diptera: Culicidae).

Blood-engorged Coquillettidia perturbans, Psorophora ferox, Culex, Culiseta, and Aedes mosquitoes were collected principally by sweep net from salt marsh and woodland habitats in Connecticut. Of the 570 mosquitoes tested, precipitin tests identified the origins of 517 blood meals and revealed distinct host feeding patterns. Aedes mosquitoes fed chiefly on mammals; A. abserratus, A. cantator, and A. vexans showed selectivity for cattle and (or) horses. A. cantator also obtained blood from avian hosts and, in some instances, showed mixed passerine-mammal blood meals. These findings increase the vector potential of this salt marsh mosquito for eastern equine encephalomyelitis virus. Feedings on deer by A. abserratus suggest potential involvement of this mosquito in the transmission of certain subtypes of California encephalitis. Culex-pipiens, C. restuans, Culiseta melanura, and Cs. morsitans dyari acquired blood almost exclusively from passeriform birds.

Aedes

Experimental transmission of Sarcocystis from icterid birds to sparrows and canaries by sporocysts from the opossum.

Cowbirds (Molothrus ater) and grackles (Cassidix mexicanus) infected with muscle cysts of Sarcocystis were fed to opposums (Didelphis virginiana) and fecal sporocysts from the latter were given to sparrows (Passer domesticus, Family Ploceidae), canaries (Serinus canarius, Family Fringillidae) and ducks (Anas platyrhynchos, Family Anatidae). Asexual parasites were found in the endothelium of sparrows and canaries but not in ducks. When birds were kept 10 weeks or more after infection, muscle cysts were found grossly and microscopically in the majority of sparrows, and in 1 canary, but not in ducks. Muscle zoites were found in digests of all sparrows and canaries but not in that of ducks. Metrocytes and forms dividing by endodyogeny also were found in the digest. Thus, avian Sarcocystis was transmitted experimentally from 2 genera of 1 family (Icteridae) to 2 different families of passerine intermediate hosts by sporocysts from the definitive host. This is the broadest intermediate host spectrum known for a species of Sarcocystis.

Animals