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The increasing involvement of wild waterfowl in H5 Highly Pathogenic Avian Influenza Virus (HPAIV) circulation continues to pose a threat to animal and public health worldwide. In winter 2020–2021, two field surveillance activities were carried out on a weekly basis, through virological and serological analyses, in 823 hunted and 521 trapped migratory aquatic birds in northeast Italy. Sixty Eurasian teals were recaptured several times, which allowed us to follow the progression of the HPAI H5 infection in naturally infected wild waterfowl. Oropharyngeal, cloacal, and feather swabs (OS, CS and FS) were collected from each duck and tested by real time rRT-PCR Type A influenza. The identified viruses were characterized and pathotyped by sequencing. Several viruses belonging to three different HPAI H5 subtypes were detected: H5N8, H5N5, and H5N1. High prevalence of infection with HPAI H5 clade 2.3.4.4b during November–December 2020 (up to 27.1%) was observed in captured Eurasian teals, while infection rates in hunted dabbling ducks, mainly Eurasian wigeons, showed the highest prevalence of infection in November 2020 (8.9%) and January 2021 (10.2%). All HPAI positive birds were also clinically healthy when recaptured weeks apart. The OS and FS showed the highest detection efficiency of HPAIV. Our results highlight that HPAI passive surveillance should be complemented by a targeted active surveillance to more efficiently detect novel HPAI viruses.

The Bare-faced Ibis is a gregarious bird found in South America. In the face of a rapid expansion in a southern state in Brazil and a lack of detailed information about the species' reproduction, two breeding areas were studied weekly for two consecutive seasons. We registered numbers and characteristics of adults, nests, eggs, and chicks, calculated success estimates, and assessed nest installation preferences. Chick morphology, age, and measurements served to define five growth stages. The species used the margins of two close rivers in Santa Catarina to build nests, lay eggs, and develop parental care between August and January, with success estimates as expected for the family. We observed a preference for the eastern margin of the rivers and proximity to bridges for nest installation, where the muddy margin provides resources for juvenile feeding. Two colony models were defined based on the near or far installation of the nests. Variations in egg and chick dimensions indicated higher adult investment in the second season, with negative allometric growth of the bill and tarsus of chicks related to weight. The results presented suggest favorable conditions for the species’ reproduction in the region, a considerable factor in understanding its expansion in recent years.

Aquatic bird bornavirus 1 (ABBV-1) is a neurotropic virus that infects waterfowls, resulting in persistent infection. Experimental infection showed that both Muscovy ducks and chickens support persistent ABBV-1 infection in the central nervous system (CNS), up to 12 weeks post-infection (wpi), without the development of clinical disease. The aim of the present study was to describe the transcriptomic profiles in the brains of experimentally infected Muscovy ducks and chickens infected with ABBV-1 at 4 and 12 wpi. Transcribed RNA was sequenced by next-generation sequencing and analyzed by principal component analysis (PCA) and differential gene expression. The functional annotation of differentially expressed genes was evaluated by gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. The PCA showed that the infected ducks sampled at both 4 and 12 wpi clustered separately from the controls, while only the samples from the chickens at 12 wpi, but not at 4 wpi, formed a separate cluster. In the ducks, more genes were differentially expressed at 4 wpi than 12 wpi, and the majority of the highly differentially expressed genes (DEG) were upregulated. On the other hand, the infected chickens had fewer DEGs at 4 wpi than at 12 wpi, and the majority of those with high numbers of DEGs were downregulated at 4 wpi and upregulated at 12 wpi. The functional annotation showed that the most enriched GO terms were immune-associated in both species; however, the terms associated with the innate immune response were predominantly enriched in the ducks, whereas the chickens had enrichment of both the innate and adaptive immune response. Immune-associated pathways were also enriched according to the KEGG pathway analysis in both species. Overall, the transcriptomic analysis of the duck and chicken brains showed that the main biological responses to ABBV-1 infection were immune-associated and corresponded with the levels of inflammation in the CNS.

Background Aquatic bird bornavirus 1 (ABBV-1) has been associated with neurological diseases in wild waterfowls. In Canada, presence of ABBV-1 was demonstrated by RT-qPCR and immunohistochemistry in tissues of waterfowls with history of neurological disease and inflammation of the central and peripheral nervous tissue, although causation has not been proven by pathogenesis experiments, yet. To date, in vitro characterization of ABBV-1 is limited to isolation in primary duck embryo fibroblasts. The objectives of this study were to describe isolation of ABBV-1 in primary duck embryonic fibroblasts (DEF), and characterize replication in DEF and three immortalized avian fibroblast cell lines (duck CCL-141, quail QT-35, chicken DF-1) in order to evaluate cellular permissivity and identify suitable cell lines for routine virus propagation. Methods The virus was sequenced, and phylogenetic analysis performed on a segment of the N gene coding region. Virus spread in cell cultures, viral RNA and protein production, and titres were evaluated at different passages using immunofluorescence, RT-qPCR, western blotting, and tissue culture dose 50% (TCID 50 ) assay, respectively. Results The isolated ABBV-1 showed 97 and 99% identity to European ABBV-1 isolate AF-168 and North American ABBV-1 isolates 062-CQ and CG-N1489, and could infect and replicate in DEF, CCL-141, QT-35 and DF-1 cultures. Viral RNA was detected in all four cultures with highest levels observed in DEF and CCL-141, moderate in QT-35, and lowest in DF-1. N protein was detected in western blots from infected DEF, CCL-141 and QT-35 at moderate to high levels, but minimally in infected DF-1. Infectious titre was highest in DEF (between approximately 10 5 to 10 6 FFU / 10 6 cells). Regarding immortalized cell lines, CCL-141 showed the highest titre between approximately 10 4 to 10 5 FFU / 10 6 cells. DF-1 produced minimal infectious titre. Conclusions This study confirms the presence of ABBV-1 among waterfowl in Canada and reported additional in vitro characterization of this virus in different avian cell lines. ABBV-1 replicated to highest titre in DEF, followed by CCL-141 and QT-35, and poorly in DF-1. Our results showed that CCL-141 can be used instead of DEF for routine ABBV-1 production, if a lower titre is an acceptable trade-off for the simplicity of using immortalized cell line over primary culture.

The present study aimed to investigate the structure, composition and diversity of the over-wintering aquatic bird community of Poyang Lake, including Poyang Lake National Nature Reserve (PNNR), Nanji National Nature Reserve (NNNR) and Duchang Provincial Nature Reserve (DPNR), China. After the preliminary survey, birds surveyed from vantage points at each study site between the years 2016 and 2020 in the winter season. A total of 58 bird species belonging to nine orders and 13 families were observed. The study showed variation in effective species numbers (Species richness, Shannon’s diversity and Simpson’s diversity) among the three study sites and the survey years. Nanji National Nature Reserve had the highest avian diversity, whereas Duchang Provincial Nature Reserve had the lowest. Globally threatened bird species, Siberian Crane (critically endangered), Oriental Stork (endangered), found in our study sites. However, the current management practices of the nature reserve and conservation of this globally threatened bird species are inadequate, especially of Duchang Provincial Nature Reserve. Therefore, for long term conservation of birds in these areas, it needs continuing intentional improvement of the sites and awareness creation to the local community.

Bornaviruses are a diverse family of viruses infecting various hosts, including birds. Aquatic bird bornavirus 1 (ABBV-1) and aquatic bird bornavirus 2 (ABBV-2) have been found in wild waterfowl but data on their prevalence are scarce. To gain knowledge on the occurrence of ABBVs in Poland, samples originating from dead birds of the Anseriformes order collected in 2016–2021 were tested with a real time RT-PCR method targeting the ABBVs genome. A total of 514 birds were examined, including 401 swans, 96 ducks and 17 geese. The presence of ABBV-1 RNA was detected in 52 swans (10.1% of all tested birds) from 40 different locations. No positive results were obtained for ducks and geese. Sequences of about 2300 bases were generated for 18 viruses and phylogenetic analysis was performed. A relatively low genetic diversity of the examined ABBV-1 strains was observed as all were gathered in a single cluster in the phylogenetic tree and the minimum nucleotide identity was 99.14%. The Polish strains were closely related to ABBV-1 identified previously in Denmark and Germany, but a limited number of sequences from Europe hinders the drawing of conclusions about interconnections between Polish and other European ABBVs. The results of the present study provide new insights into the distribution and genetic characteristics of ABBVs in wild birds in Europe.

We studied the effects of water level in the Danube River on waterbird species abundance and species assemblages along 83 km from the border of Hungary and Slovakia. 651,622 birds of 62 species were counted between 1992 and 2009. The dominant species was Mallard Anas platyrhynchos contributing 436,198 individuals (66.9% of all observations). Throughout the year total numbers of individuals and species richness were negatively correlated with local water level registered on the same day throughout the year. Shannon diversity indices were generally positively correlated with water levels. During floods, dabbling ducks, especially the dominant Anas platyrhynchos , dispersed to peripheral waters, where they are less likely to be counted, and diving ducks ( Bucephala clangula, Aythya fuligula and Mergus spp.) left the area entirely, because fast flowing, highly turbid waters reduced local feeding efficiency. Abundance of most waterbird species decreased with elevated water levels. High water levels remove distinctive microhabitats (gravel banks, paved riverbanks and shoals), and create unfavourable conditions of high water velocity and turbidity. Retention of high water levels as a result of damming of the Danube creates long-term conditions similar to natural flooding effects. In our opinion, further manipulation of Danube water levels is likely to reduce waterbird richness and abundance and should be subject to appropriate environmental impact assessment.

Highly pathogenic avian influenza (HPAI) A/H5N1 viruses (lineage 2.3.4.4b) are rapidly invading the Americas, threatening wildlife, poultry, and potentially evolving into the next global pandemic. In November 2022 HPAI arrived in Peru, triggering massive pelican and sea lion die-offs. We report genomic characterization of HPAI/H5N1 in five species of marine mammals and seabirds (dolphins, sea lions, sanderlings, pelicans and cormorants). Peruvian viruses belong to lineage 2.3.4.4b, but they are 4:4 reassortants where 4 genomic segments (PA, HA, NA and MP) position within the Eurasian lineage that initially entered North America from Eurasia, while the other 4 genomic segments (PB2, PB1, NP and NS) position within the American lineage (clade C) that circulated in North America. These viruses are rapidly accruing mutations, including mutations of concern, that warrant further examination and highlight an urgent need for active local surveillance to manage outbreaks and limit spillover into other species, including humans. Highly pathogenic avian influenza (HPAI) A/H5N1 has recently emerged in the Americas and has been implicated in mass die-off events of pelicans and sea lions. Here, the authors report sampling and characterisation of HPAI A/H5N1 genomes from five marine mammal and seabird species in Peru.