Spatio-temporal dynamics and aetiology of proliferative leg skin lesions in wild British finches

Proliferative leg skin lesions have been described in wild finches in Europe although there have been no large-scale studies of their aetiology or epizootiology to date. Firstly, disease surveillance, utilising public reporting of observations of live wild finches was conducted in Great Britain (GB) and showed proliferative leg skin lesions in chaffinches (Fringilla coelebs) to be widespread. Seasonal variation was observed, with a peak during the winter months. Secondly, pathological investigations were performed on a sample of 39 chaffinches, four bullfinches (Pyrrhula pyrrhula), one greenfinch (Chloris chloris) and one goldfinch (Carduelis carduelis) with proliferative leg skin lesions and detected Cnemidocoptes sp. mites in 91% (41/45) of affected finches and from all species examined. Fringilla coelebs papillomavirus (FcPV1) PCR was positive in 74% (23/31) of birds tested: a 394 base pair sequence was derived from 20 of these birds, from all examined species, with 100% identity to reference genomes. Both mites and FcPV1 DNA were detected in 71% (20/28) of birds tested for both pathogens. Histopathological examination of lesions did not discriminate the relative importance of mite or FcPV1 infection as their cause. Development of techniques to localise FcPV1 within lesions is required to elucidate the pathological significance of FcPV1 DNA detection.We thank the members of the public and BTO Garden BirdWatch participants who reported garden bird morbidity and mortality incidents and our colleagues, Katie Beckmann, Shaheed Macgregor, Ricardo Castro Cesar de Sa, Lydia Franklinos and Tim Hopkins from the Zoological Society of London; Kirsi Peck from the Royal Society for the Protection of Birds; BTO staff members in the Garden BirdWatch team; the staff at Abbey Veterinary Services and the Animal & Plant Health Agency (Daniel Hicks, Richard Irvine, Alejandro Núñez and Scott Reid) for their assistance with this investigation. This work was financially supported by the following organisations; Birdcare Standards Association, British Trust for Ornithology, British Veterinary Association Animal Welfare Foundation, CJ Wildbird Foods, Cranswick Pet Products, UK Department for the Environment Food & Rural Affairs and Welsh Government through the Animal & Plant Health Agency’s Diseases of Wildlife Scheme Scanning Surveillance Programme (Project ED1600), Esmée Fairbairn Foundation, Gardman Ltd, Institute of Zoology, Royal Society for the Protection of Birds and the Universities Federation for Animal Welfare. RAJW was supported by the Moncloa of Excellence PICATA programme and Crafoord Foundation Sweden (grant number 20160971). Molecular and sequencing costs were funded by the Spanish Ministry of Science and Innovation, (Ref: CGL2013-41642-P/BOS)

Evidence for Ljungan virus specific antibodies in humans and rodents, Finland.

Objectives Ljungan virus (LV) belongs to the genus Parechovirus. Human parechoviruses (HPeV) are common viruses causing diarrhea and gastroenteritis, and the first infection is commonly faced during the early childhood and 90% of humans acquire HPeV antibodies by the age of two. Ljungan virus, however, is known as a rodent-borne parechovirus first isolated (LV 87-012) near Ljungan river in Sweden from a wild bank vole (Myodes glareolus). Puumala hantavirus causes Nephropatia epidemica (NE) in humans and these cases are fairly common in Finland comprising approximately 800-3000 suspected NE cases per year depending somewhat on the size of rodent population. However, the abundance of Finnish patients diagnosed with NE, i.e., with a history of rodent contact, makes any connection between human disease and Ljungan virus infection likely to be found in the country. With this study, we sought to find evidence of Ljungan virus in Finnish humans and rodents. We aimed to develop a reliable method for serological screening of LV in humans and rodents, and to confirm the specificity of this method. Methods Initially, neutralization assays were designed and carried out followed by an immunofluorescent assay (IFA) for serology screening. The IFA and neutralization assays were set up using Vero-cells infected with Ljungan virus strain 145SLG kindly provided by Conny Tolf et al. (Uppsala University, Uppsala, Sweden). Serological assays were used for detecting Ljungan virus antibodies both in humans and rodents. Furthermore, 8 human sera used in the Ljungan virus neutralization assay were cross-checked for titres of neutralizing antibodies to six different human parechoviruses (HPeV 1-6), that are closely related to Ljungan virus. Neutralization effect of the antibodies to 42 different human picornaviruses against Ljungan virus (145SLG) was also studied. Results and Conclusion The study is ongoing, but preliminary data indicates that we have LV specific antibodies in Finland. Twenty-six sera out of 41 human serums were not able to neutralize Ljungan 145SLG virus. However, 15 human serum samples neutralized the Ljungan 145SLG virus. Wide titer range of the HPeV 1-6 antibodies were also detected in eight human serums studied in more closely. The preliminary IFA results are also supporting the neutralization results. In total, 8 out of the 50 (16%, 95% CI: 8.1–28.8%) rodent samples (Konnevesi, Finland, year 2008) were Ljungan antibody positive. Our data is first evidence of human Ljungan virus infections in Finland both in humans and rodents. This data can be used as a stepping stone for further studies of Ljungan virus: the broad seroprevalence study both in humans and rodents, and to investigate the role of Ljungan virus infection in Finland

Spatio-temporal dynamics and aetiology of proliferative leg skin lesions in wild British finches

This work was financially supported by the following organisations; Birdcare Standards Association, British Trust for Ornithology, British Veterinary Association Animal Welfare Foundation, CJ Wildbird Foods, Cranswick Pet Products, UK Department for the Environment Food & Rural Affairs and Welsh Government through the Animal & Plant Health Agency’s Diseases of Wildlife Scheme Scanning Surveillance Programme (Project ED1600), Esmée Fairbairn Foundation, Gardman Ltd, Institute of Zoology, Royal Society for the Protection of Birds and the Universities Federation for Animal Welfare. RAJW was supported by the Moncloa of Excellence PICATA programme and Crafoord Foundation Sweden (grant number 20160971). Molecular and sequencing costs were funded by the Spanish Ministry of Science and Innovation, (Ref: CGL2013-41642-P/BOS).Proliferative leg skin lesions have been described in wild finches in Europe although there have been no large-scale studies of their aetiology or epizootiology to date. Firstly, disease surveillance, utilising public reporting of observations of live wild finches was conducted in Great Britain (GB) and showed proliferative leg skin lesions in chaffinches (Fringilla coelebs) to be widespread. Seasonal variation was observed, with a peak during the winter months. Secondly, pathological investigations were performed on a sample of 39 chaffinches, four bullfinches (Pyrrhula pyrrhula), one greenfinch (Chloris chloris) and one goldfinch (Carduelis carduelis) with proliferative leg skin lesions and detected Cnemidocoptes sp. mites in 91% (41/45) of affected finches and from all species examined. Fringilla coelebs papillomavirus (FcPV1) PCR was positive in 74% (23/31) of birds tested: a 394 base pair sequence was derived from 20 of these birds, from all examined species, with 100% identity to reference genomes. Both mites and FcPV1 DNA were detected in 71% (20/28) of birds tested for both pathogens. Histopathological examination of lesions did not discriminate the relative importance of mite or FcPV1 infection as their cause. Development of techniques to localise FcPV1 within lesions is required to elucidate the pathological significance of FcPV1 DNA detection.Depto. de Genética, Fisiología y MicrobiologíaFac. de Ciencias BiológicasTRUEpu

Birds and Viruses at a Crossroad : Surveillance of Influenza A Virus in Portuguese Waterfowl

During recent years, extensive amounts of data have become available regarding influenza A virus (IAV) in wild birds in northern Europe, while information from southern Europe is more limited. Here, we present an IAV surveillance study conducted in western Portugal 2008-2009, analyzing 1653 samples from six different species of waterfowl, with the majority of samples taken from Mallards (Anas platyrhynchos). Overall 4.4% of sampled birds were infected. The sampling results revealed a significant temporal variation in the IAV prevalence, including a pronounced peak among predominantly young birds in June, indicating that IAV circulate within breeding populations in the wetlands of western Portugal. The H10N7 and H9N2 subtypes were predominant among isolated viruses. Phylogenetic analyses of the hemagglutinin and neuraminidase sequences of H10N7, H9N2 and H11N3 virus showed that sequences from Portugal were closely related to viral sequences from Central Europe as well as to IAVs isolated in the southern parts of Africa, reflecting Portugal's position on the European-African bird migratory flyway. This study highlights the importance of Portugal as a migratory crossroad for IAV, connecting breeding stationary waterfowl with birds migrating between continents which enable transmission and spread of IAV