351 lines
41 KiB
XML
351 lines
41 KiB
XML
<document id="4FF371973209952837588A72FFF4E865" ID-DOI="10.1016/j.ijppaw.2023.100895" ID-ISSN="2213-2244" ID-PMC="PMC10767487" ID-PubMed="38187443" ID-Zenodo-Dep="10933806" IM.bibliography_approvedBy="carolina" IM.illustrations_approvedBy="carolina" IM.materialsCitations_approvedBy="felipe" IM.metadata_approvedBy="felipe" IM.tables_approvedBy="carolina" IM.taxonomicNames_approvedBy="carolina" IM.treatments_approvedBy="carolina" checkinTime="1712341721380" checkinUser="felipe" docAuthor="Cigler, P., More, G., Bize, P., Meier, C. M., Frey, C. F., Basso, W. & Keller, S." docDate="2024" docId="BE4A562FFFEBFF93B23BFBEF9107FE81" docLanguage="en" docName="IntJParasitolParasitesWildlife.23.100895.pdf" docOrigin="International Journal for Parasitology: Parasites and Wildlife 23" docSource="http://dx.doi.org/10.1016/j.ijppaw.2023.100895" docStyle="DocumentStyle:77CC2CF2FBA7BCD129DD16D0D4F96931.2:IntJParasitolParasitesWildlife.2012-.journal_article" docStyleId="77CC2CF2FBA7BCD129DD16D0D4F96931" docStyleName="IntJParasitolParasitesWildlife.2012-.journal_article" docStyleVersion="2" docTitle="Trypanosoma Gruby" docType="treatment" docVersion="4" masterDocId="42732E57FFE9FF96B109FFE09205FFEE" masterDocTitle="Trypanosomiasis: An emerging disease in Alpine swift (Tachymarptis melba) nestlings in Switzerland?" masterLastPageNumber="100895" masterPageNumber="100895" pageNumber="100895" updateTime="1713820453842" updateUser="ExternalLinkService" zenodo-license-document="CC-BY-NC-ND-4.0">
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<mods:title id="BC674A0CF744A5DFBF6997DC2042BEE6">Trypanosomiasis: An emerging disease in Alpine swift (Tachymarptis melba) nestlings in Switzerland?</mods:title>
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<mods:namePart id="CFA00DB6AC5536F8832C2DA531A16EA2">Cigler, P.</mods:namePart>
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<mods:affiliation id="99580B7681A8BCF7A93B19DD90FD564E">Institute for Fish and Wildlife Health, University of Bern, L ¨ anggassstrasse 122, 3012 Bern, Switzerland</mods:affiliation>
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<mods:nameIdentifier id="0E80335FA772DF9781D6441AAC43FCD6" type="email">pia.cigler@unibe.ch</mods:nameIdentifier>
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<mods:namePart id="71A57C4A042FCE7F7ACFC126D9587681">More, G.</mods:namePart>
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<mods:affiliation id="118755E8A6855F10E9E1FA4FA4E98436">Institute of Parasitology, University of Bern, L ¨ anggassstrasse 122, 3012 Bern, Switzerland</mods:affiliation>
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<mods:namePart id="F0FEB90A728A4C713D97E85A1392EC9F">Bize, P.</mods:namePart>
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<mods:affiliation id="B32C0D8BAAA61BBE3CA25D1DCCF6C1AE">Swiss Ornithological Institute, Seerose 1, 6204 Sempach, Switzerland</mods:affiliation>
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<mods:namePart id="983B4F0293FD753599D43D3B41676FA8">Meier, C. M.</mods:namePart>
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<mods:affiliation id="D180E7B9FC9EDC9418B44018DBE0A3B0">Swiss Ornithological Institute, Seerose 1, 6204 Sempach, Switzerland</mods:affiliation>
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<mods:namePart id="17EFAE56E1DAF305525BF84BDCE5FD1C">Frey, C. F.</mods:namePart>
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<mods:affiliation id="176DF90986C0BD67F668ED5DB99C958C">Institute of Parasitology, University of Bern, L ¨ anggassstrasse 122, 3012 Bern, Switzerland</mods:affiliation>
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<mods:namePart id="57450ED709CD29682B61BE8D2DB63AC3">Basso, W.</mods:namePart>
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<mods:affiliation id="0AA29ED87D20CB6FCD2D1FFD6B313647">Institute of Parasitology, University of Bern, L ¨ anggassstrasse 122, 3012 Bern, Switzerland</mods:affiliation>
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<mods:namePart id="C6EBE5CBDB458723DD93BCBC14B1DA4B">Keller, S.</mods:namePart>
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<mods:affiliation id="FA2D0D9A61E9102AB321263634C7FEC7">Institute for Fish and Wildlife Health, University of Bern, L ¨ anggassstrasse 122, 3012 Bern, Switzerland</mods:affiliation>
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<mods:title id="21F7001B53874A851D2894816FF84690">International Journal for Parasitology: Parasites and Wildlife</mods:title>
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<mods:date id="0E28A3C69AB208A447A3C65FB0771446">2024</mods:date>
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<mods:number id="B1A8E092188A0CE5D543E68168B6470D">2024-04-30</mods:number>
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<mods:number id="A2411D719752BE90D02EEA91AA47E831">23</mods:number>
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<mods:identifier id="D4A45E5B7A067480F50D1D13E302B014" type="DOI">10.1016/j.ijppaw.2023.100895</mods:identifier>
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<mods:identifier id="D4F3A6423170A272AE1B477313834263" type="ISSN">2213-2244</mods:identifier>
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<treatment id="BE4A562FFFEBFF93B23BFBEF9107FE81" ID-DOI="http://doi.org/10.5281/zenodo.11043549" ID-Zenodo-Dep="11043549" LSID="urn:lsid:plazi:treatment:BE4A562FFFEBFF93B23BFBEF9107FE81" httpUri="http://treatment.plazi.org/id/BE4A562FFFEBFF93B23BFBEF9107FE81" lastPageId="5" pageId="2">
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<subSubSection id="7EF9B4B2FFEBFF94B23BFBEF9690FBCD" box="[818,1173,1039,1059]" pageId="2" type="nomenclature">
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<paragraph id="365CE739FFEBFF94B23BFBEF9690FBCD" blockId="2.[818,1488,1039,1339]" box="[818,1173,1039,1059]" pageId="2">
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<heading id="6D145055FFEBFF94B23BFBEF9690FBCD" box="[818,1173,1039,1059]" fontSize="36" level="2" pageId="2" reason="8">
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<emphasis id="04973B2BFFEBFF94B23BFBEF9690FBCD" bold="true" box="[818,1173,1039,1059]" italics="true" pageId="2">
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2.4.1.
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<taxonomicName id="F1E39CBAFFEBFF94B27DFBEF91E8FBCD" ID-CoL="64874" authority="Gruby, 1843" authorityName="Gruby" box="[884,1005,1039,1059]" class="Kinetoplastea" family="Trypanosomatidae" genus="Trypanosoma" kingdom="Protozoa" order="Trypanosomatida" pageId="2" phylum="Euglenozoa" rank="genus">Trypanosoma</taxonomicName>
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sp. morphometrics
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</emphasis>
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</heading>
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</paragraph>
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</subSubSection>
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<subSubSection id="7EF9B4B2FFEBFF94B258FBCC97CAFAD4" pageId="2" type="discussion">
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<paragraph id="365CE739FFEBFF94B258FBCC97CAFAD4" blockId="2.[818,1488,1039,1339]" pageId="2">
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Trypomastigotes (between 6 and 15 per sample) found in three samples from 2022 were measured. The smears were evaluated using a Nikon Eclipse
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<emphasis id="04973B2BFFEBFF94B2B1FB8391CEFB99" bold="true" box="[952,971,1123,1143]" italics="true" pageId="2">Ci</emphasis>
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microscope and photographed with a calibrated Nikon Camera (model DFK 23
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<collectionCode id="50F27FFCFFEBFF94B503FB60962FFB7D" box="[1034,1066,1152,1171]" country="Papua New Guinea" name="University of Papua and New Guinea" pageId="2">UP</collectionCode>
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031). Measurements were taken with the NIS- Elements D software (Version 5.02.03 64 bit) and included: total length without flagellum (PA), posterior end to centre of kinetoplast (
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<collectionCode id="50F27FFCFFEBFF94B4AFFB5897C3FB25" box="[1446,1478,1208,1227]" pageId="2">PK</collectionCode>
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), posterior end to centre of nucleus (
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<collectionCode id="50F27FFCFFEBFF94B57AFB349691FB09" box="[1139,1172,1236,1255]" pageId="2">PN</collectionCode>
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), centre of nucleus to anterior end (NA), centre of kinetoplast to centre of nucleus (
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<collectionCode id="50F27FFCFFEBFF94B5FFFB10971CFAED" box="[1270,1305,1264,1283]" pageId="2">KN</collectionCode>
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), body width at the centre of the nucleus (
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<collectionCode id="50F27FFCFFEBFF94B505FAEB9637FAF0" box="[1036,1074,1291,1310]" pageId="2">BW</collectionCode>
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), length of free flagellum (
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<collectionCode id="50F27FFCFFEBFF94B43EFAEB9756FAF0" box="[1335,1363,1291,1310]" pageId="2">FF</collectionCode>
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), area of the trypomastigote (
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<collectionCode id="50F27FFCFFEBFF94B2C3FAC791ECFAD4" box="[970,1001,1319,1338]" pageId="2">AT</collectionCode>
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) and area of the nucleus (
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<collectionCode id="50F27FFCFFEBFF94B5DEFAC796FCFAD4" box="[1239,1273,1319,1338]" pageId="2">AN</collectionCode>
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) (
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<bibRefCitation id="52729AC8FFEBFF94B402FAC897C4FAD4" author="Bennett" box="[1291,1473,1319,1339]" etAl="et al." firstAuthor="Bennett" pageId="2" pagination="263 - 263" refId="ref7885" refString="Bennett, G. F., Earle, R., Squires-Parsons, D., 1994. Trypanosomes of some sub-Saharan birds. Onderstepoort J. Vet. Res. 61,, 263 - 263." type="journal article" year="1994">Bennett et al., 1994</bibRefCitation>
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).
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</paragraph>
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||
</subSubSection>
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<subSubSection id="7EF9B4B2FFEBFF93B23BFABF9107FE81" lastPageId="5" pageId="2" type="description">
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<paragraph id="365CE739FFEBFF94B23BFABF919EFA9C" blockId="2.[818,923,1375,1394]" box="[818,923,1375,1394]" pageId="2">
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<heading id="6D145055FFEBFF94B23BFABF919EFA9C" bold="true" box="[818,923,1375,1394]" fontSize="36" level="1" pageId="2" reason="1">
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<emphasis id="04973B2BFFEBFF94B23BFABF919EFA9C" bold="true" box="[818,923,1375,1394]" pageId="2">3. Results</emphasis>
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</heading>
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</paragraph>
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<paragraph id="365CE739FFEBFF94B23BFA769604FA44" blockId="2.[818,1025,1430,1450]" box="[818,1025,1430,1450]" pageId="2">
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<heading id="6D145055FFEBFF94B23BFA769604FA44" box="[818,1025,1430,1450]" fontSize="36" level="2" pageId="2" reason="3">
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<emphasis id="04973B2BFFEBFF94B23BFA769604FA44" bold="true" box="[818,1025,1430,1450]" italics="true" pageId="2">3.1. Field observations</emphasis>
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</heading>
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</paragraph>
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<paragraph id="365CE739FFEBFF94B258FA2F971AF94B" blockId="2.[818,1488,1487,1897]" pageId="2">
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The affected nestlings showed pale coloration of mucous membranes and feet, alongside moderate to severe bruising across the body. These signs were observed earliest in nestlings aged 14–20 days, although most observations were in nestlings aged 20–45 days. Other changes in the form of primary feather abnormalities, including stunted growth, retained feather shafts and/or blood keels, were observed in nestlings aged 40 days and over (
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<figureCitation id="AED8FBBCFFEBFF94B529F996965DF967" box="[1056,1112,1654,1673]" captionStart="Fig" captionStartId="3.[100,130,726,743]" captionTargetBox="[209,661,149,697]" captionTargetId="figure-496@3.[208,662,148,698]" captionTargetPageId="3" captionText="Fig. 3. Missing (left wing) and poor quality (right wing) primary feathers on a 45-day-old nestling." figureDoi="http://doi.org/10.5281/zenodo.10933812" httpUri="https://zenodo.org/record/10933812/files/figure.png" pageId="2">Fig. 3</figureCitation>
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). However, as these changes were not noted consistently, the exact numbers are unknown.
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</paragraph>
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<paragraph id="365CE739FFEBFF94B258F94E96DCF8FA" blockId="2.[818,1488,1487,1897]" pageId="2">A considerable louse fly burden was described on both healthy and affected nestlings, with a mean of 5.5 ± 0.2 per nestling throughout development (690 observations). The highest burden on an individual nestling observed in 2022 was 39 louse flies.</paragraph>
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<paragraph id="365CE739FFEBFF94B258F8FD975BF887" blockId="2.[818,1488,1487,1897]" pageId="2">
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The numbers of fledged, deceased or missing (but presumed dead) nestlings, as well as the average age and weight at the time of death were evaluated for colonies A, B and C and compiled in
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<tableCitation id="7B61D282FFEBFF94B418F8B5975BF887" box="[1297,1374,1877,1897]" captionStart-0="Table 1" captionStart-1="Table 2" captionStart-2="Table 3" captionStartId-0="3.[100,150,828,844]" captionStartId-1="5.[100,150,1834,1850]" captionStartId-2="6.[100,150,993,1009]" captionTargetPageId-0="3" captionTargetPageId-1="5" captionTargetPageId-2="6" captionText-0="Table 1 Summary of the number of nestlings ringed, fledged and deceased or missing in colonies A, B and C, including median, minimum, and maximum age and weight at death. Ref. * = median (min/max)." captionText-1="Table 2 Average counts of 10 high-power-field counts (HPF) 400× magnification of positive and negative nestlings, with Standard deviation and range in brackets." captionText-2="Table 3 Average morphometric measurements of 36 trypomastigotes observed in 2022 from three 45-day-old Alpine swift nestlings. SD: Standard deviation." httpUri-0="http://table.plazi.org/id/629CB7B1FFEAFF95B16DFCDC93A2FC77" httpUri-1="http://table.plazi.org/id/629CB7B1FFECFF93B16DF8CA978EF8BA" httpUri-2="http://table.plazi.org/id/629CB7B1FFEFFF90B16DFC0190B5FBCB" pageId="2" tableUuid-0="629CB7B1FFEAFF95B16DFCDC93A2FC77" tableUuid-1="629CB7B1FFECFF93B16DF8CA978EF8BA" tableUuid-2="629CB7B1FFEFFF90B16DFC0190B5FBCB">Table 1.</tableCitation>
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</paragraph>
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<caption id="629CB7B1FFEAFF95B16DFD369315FCEE" ID-DOI="http://doi.org/10.5281/zenodo.10933812" ID-Zenodo-Dep="10933812" httpUri="https://zenodo.org/record/10933812/files/figure.png" pageId="3" startId="3.[100,130,726,743]" targetBox="[209,661,149,697]" targetPageId="3" targetType="figure">
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<paragraph id="365CE739FFEAFF95B16DFD369315FCEE" blockId="3.[100,770,725,768]" pageId="3">
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<emphasis id="04973B2BFFEAFF95B16DFD369298FD09" bold="true" box="[100,157,726,743]" pageId="3">Fig. 3.</emphasis>
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Missing (left wing) and poor quality (right wing) primary feathers on a 45-day-old nestling.
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||
</paragraph>
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||
</caption>
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||
<caption id="629CB7B1FFEAFF95B16DFCDC93A2FC77" ID-Table-UUID="629CB7B1FFEAFF95B16DFCDC93A2FC77" httpUri="http://table.plazi.org/id/629CB7B1FFEAFF95B16DFCDC93A2FC77" pageId="3" startId="3.[100,150,828,844]" targetBox="[116,753,941,1105]" targetIsTable="true" targetPageId="3" targetType="table">
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<paragraph id="365CE739FFEAFF95B16DFCDC92A2FCA3" blockId="3.[100,770,828,921]" box="[100,167,828,845]" pageId="3">
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<emphasis id="04973B2BFFEAFF95B16DFCDC92A2FCA3" bold="true" box="[100,167,828,845]" pageId="3">Table 1</emphasis>
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</paragraph>
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||
<paragraph id="365CE739FFEAFF95B16DFCB593A2FC77" blockId="3.[100,770,828,921]" pageId="3">Summary of the number of nestlings ringed, fledged and deceased or missing in colonies A, B and C, including median, minimum, and maximum age and weight at death. Ref. * = median (min/max).</paragraph>
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</caption>
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<paragraph id="365CE739FFEAFF95B055FC4D908EFBBE" pageId="3">
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<table id="44E31599FFEA0069B17DFC4D90F4FBBF" box="[116,753,941,1105]" gridcols="4" gridrows="7" pageId="3">
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<tr id="88D3E57BFFEA0069B17DFC4D90F4FC52" box="[116,753,941,956]" gridrow="0" pageId="3" rowspan-0="1">
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<th id="CB028C07FFEA0069B055FC4D93A4FC52" box="[348,417,941,956]" gridcol="1" gridrow="0" pageId="3">Colony A</th>
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<th id="CB028C07FFEA0069B0C8FC4D904FFC52" box="[449,586,941,956]" gridcol="2" gridrow="0" pageId="3">Colony B</th>
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<th id="CB028C07FFEA0069B363FC4D90F4FC52" box="[618,753,941,956]" gridcol="3" gridrow="0" pageId="3">Colony C</th>
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</tr>
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<tr id="88D3E57BFFEA0069B17DFC3090F4FC31" box="[116,753,976,991]" gridrow="1" pageId="3">
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<th id="CB028C07FFEA0069B17DFC309339FC31" box="[116,316,976,991]" gridcol="0" gridrow="1" pageId="3">Nr. of ringed nestlings</th>
|
||
<td id="CB028C07FFEA0069B055FC3093A4FC31" box="[348,417,976,991]" gridcol="1" gridrow="1" pageId="3">99</td>
|
||
<td id="CB028C07FFEA0069B0C8FC30904FFC31" box="[449,586,976,991]" gridcol="2" gridrow="1" pageId="3">74</td>
|
||
<td id="CB028C07FFEA0069B363FC3090F4FC31" box="[618,753,976,991]" gridcol="3" gridrow="1" pageId="3">95</td>
|
||
</tr>
|
||
<tr id="88D3E57BFFEA0069B17DFC0790F4FC1B" box="[116,753,999,1013]" gridrow="2" pageId="3">
|
||
<th id="CB028C07FFEA0069B17DFC079339FC1B" box="[116,316,999,1013]" gridcol="0" gridrow="2" pageId="3">Nr. of fledged nestlings</th>
|
||
<td id="CB028C07FFEA0069B055FC0793A4FC1B" box="[348,417,999,1013]" gridcol="1" gridrow="2" pageId="3">49</td>
|
||
<td id="CB028C07FFEA0069B0C8FC07904FFC1B" box="[449,586,999,1013]" gridcol="2" gridrow="2" pageId="3">54</td>
|
||
<td id="CB028C07FFEA0069B363FC0790F4FC1B" box="[618,753,999,1013]" gridcol="3" gridrow="2" pageId="3">26</td>
|
||
</tr>
|
||
<tr id="88D3E57BFFEA0069B17DFC1E90F4FBE2" box="[116,753,1022,1036]" gridrow="3" pageId="3">
|
||
<th id="CB028C07FFEA0069B17DFC1E9339FBE2" box="[116,316,1022,1036]" gridcol="0" gridrow="3" pageId="3">Nr. of deceased or missing</th>
|
||
<td id="CB028C07FFEA0069B055FC1E93A4FBE2" box="[348,417,1022,1036]" gridcol="1" gridrow="3" pageId="3">50</td>
|
||
<td id="CB028C07FFEA0069B0C8FC1E904FFBE2" box="[449,586,1022,1036]" gridcol="2" gridrow="3" pageId="3">20</td>
|
||
<td id="CB028C07FFEA0069B363FC1E90F4FBE2" box="[618,753,1022,1036]" gridcol="3" gridrow="3" pageId="3">69</td>
|
||
</tr>
|
||
<tr id="88D3E57BFFEA0069B17DFBF590F4FBCD" box="[116,753,1045,1059]" gridrow="4" pageId="3">
|
||
<th id="CB028C07FFEA0069B17DFBF59339FBCD" box="[116,316,1045,1059]" gridcol="0" gridrow="4" pageId="3">Age of death (days)*</th>
|
||
<td id="CB028C07FFEA0069B055FBF593A4FBCD" box="[348,417,1045,1059]" gridcol="1" gridrow="4" pageId="3">n/a</td>
|
||
<td id="CB028C07FFEA0069B0C8FBF5904FFBCD" box="[449,586,1045,1059]" gridcol="2" gridrow="4" pageId="3">21 (14/49)</td>
|
||
<td id="CB028C07FFEA0069B363FBF590F4FBCD" box="[618,753,1045,1059]" gridcol="3" gridrow="4" pageId="3">24 (14/49)</td>
|
||
</tr>
|
||
<tr id="88D3E57BFFEA0069B17DFBCC90F4FBD4" box="[116,753,1068,1082]" gridrow="5" pageId="3">
|
||
<th id="CB028C07FFEA0069B17DFBCC9339FBD4" box="[116,316,1068,1082]" gridcol="0" gridrow="5" pageId="3">Weight at death (g)*</th>
|
||
<td id="CB028C07FFEA0069B055FBCC93A4FBD4" box="[348,417,1068,1082]" gridcol="1" gridrow="5" pageId="3">n/a</td>
|
||
<td id="CB028C07FFEA0069B0C8FBCC904FFBD4" box="[449,586,1068,1082]" gridcol="2" gridrow="5" pageId="3">83.7 (57.7/107.7)</td>
|
||
<td id="CB028C07FFEA0069B363FBCC90F4FBD4" box="[618,753,1068,1082]" gridcol="3" gridrow="5" pageId="3">88.6 (46.8/112.2)</td>
|
||
</tr>
|
||
<tr id="88D3E57BFFEA0069B17DFBA290F4FBBF" box="[116,753,1090,1105]" gridrow="6" pageId="3">
|
||
<th id="CB028C07FFEA0069B17DFBA29339FBBF" box="[116,316,1090,1105]" gridcol="0" gridrow="6" pageId="3">Mortality (%)</th>
|
||
<td id="CB028C07FFEA0069B055FBA293A4FBBF" box="[348,417,1090,1105]" gridcol="1" gridrow="6" pageId="3">50.5</td>
|
||
<td id="CB028C07FFEA0069B0C8FBA2904FFBBF" box="[449,586,1090,1105]" gridcol="2" gridrow="6" pageId="3">27.0</td>
|
||
<td id="CB028C07FFEA0069B363FBA290F4FBBF" box="[618,753,1090,1105]" gridcol="3" gridrow="6" pageId="3">72.6</td>
|
||
</tr>
|
||
</table>
|
||
</paragraph>
|
||
<paragraph id="365CE739FFEAFF95B16DFB66902CFB74" blockId="3.[100,553,1158,1178]" box="[100,553,1158,1178]" pageId="3">
|
||
<heading id="6D145055FFEAFF95B16DFB66902CFB74" box="[100,553,1158,1178]" fontSize="36" level="2" pageId="3" reason="3">
|
||
<emphasis id="04973B2BFFEAFF95B16DFB66902CFB74" bold="true" box="[100,553,1158,1178]" italics="true" pageId="3">3.2. Post-mortem examination and histopathology</emphasis>
|
||
</heading>
|
||
</paragraph>
|
||
<paragraph id="365CE739FFEAFF95B18DFB5F972AFF30" blockId="3.[100,771,1215,1429]" lastBlockId="3.[818,1488,148,976]" pageId="3">
|
||
All necropsied nestlings (n = 5) showed identical findings on gross pathology, including extensive subcutaneous haemorrhages and pale discoloration of mucous membranes and feet. The haemorrhages were randomly distributed; however, were more prominent along the back, the top of the head, both sides of the wings and on the breast (
|
||
<figureCitation id="AED8FBBCFFEAFF95B3AEFACE90D9FAAF" box="[679,732,1326,1345]" captionStart="Fig" captionStartId="3.[100,130,1964,1981]" captionTargetBox="[339,1247,1480,1935]" captionTargetId="figure-609@3.[338,1248,1479,1936]" captionTargetPageId="3" captionText="Fig. 4. Distribution of louse flies on an Alpine swift nestling (A) compared with the distribution of bruising on post-mortem examination with plumage removed (B)." figureDoi="http://doi.org/10.5281/zenodo.10933814" httpUri="https://zenodo.org/record/10933814/files/figure.png" pageId="3">Fig. 4</figureCitation>
|
||
). In animals sampled within 24 h of death, myocardial petechiae could be observed (n = 2). Significant splenomegaly was present in all animals. All birds were in good body condition, had considerable fat reserves and full intestinal tracts. One bird had significantly shorter primary feathers on one wing, without indication of a trauma. The remainder of the plumage and the uropygial gland were unremarkable.
|
||
</paragraph>
|
||
<paragraph id="365CE739FFEAFF95B258FF07919AFDC3" blockId="3.[818,1488,148,976]" pageId="3">
|
||
Histopathology (n = 3, one from each colony) confirmed extensive areas of haemorrhage in subcutaneous tissue, reaching into the underlying musculature. The epidermis and feather follicles were without abnormality. Pectoral and wing (mainly biceps) musculature showed multifocal, interstitial, often perivascular accentuated small to medium sized infiltrations of mononuclear inflammatory cells (macrophages, lymphocytes and plasma cells,
|
||
<figureCitation id="AED8FBBCFFEAFF95B560FE6F96B3FE4C" box="[1129,1206,399,418]" captionStart="Fig" captionStartId="4.[100,130,766,783]" captionTargetBox="[207,1380,150,736]" captionTargetId="figure-311@4.[206,1381,148,738]" captionTargetPageId="4" captionText="Fig. 5. Skeletal musculature of a nestling Alpine swift showing infiltrations of mononuclear inflammatory cells (A, B) and presumably extracellular, amastigote-like structures (C). Bursa fabricii of a nestling Alpine swift with depletion of the medullary follicle with lymphocytolysis (asterisk) and a distinct epithelium (arrows) (D)." figureDoi="http://doi.org/10.5281/zenodo.10933816" httpUri="https://zenodo.org/record/10933816/files/figure.png" pageId="3">Fig. 5A</figureCitation>
|
||
/B). In two birds, multifocal haemorrhaging was observed along the epicardium and, in part, infiltrating into the underlying myocardium. Sporadically distributed throughout the myocardium, a mixed inflammation composed of macrophages, lymphocytes, plasma cells and few heterophils, was seen in all three birds.
|
||
</paragraph>
|
||
<paragraph id="365CE739FFEAFF95B258FDD7965DFD1E" blockId="3.[818,1488,148,976]" pageId="3">
|
||
In proximity to the inflammatory foci, 4–5 μm, round-to-oval structures containing two basophilic nuclear-like structures were observed. These structures resembled the amastigote stages of kinetoplastid parasites and were found primarily in skeletal and cardiac musculature, as well as sporadically dispersed amongst fat cells in connective tissue and appeared to be extracellular (
|
||
<figureCitation id="AED8FBBCFFEAFF95B55EFD2196A7FD3B" box="[1111,1186,705,725]" captionStart="Fig" captionStartId="4.[100,130,766,783]" captionTargetBox="[207,1380,150,736]" captionTargetId="figure-311@4.[206,1381,148,738]" captionTargetPageId="4" captionText="Fig. 5. Skeletal musculature of a nestling Alpine swift showing infiltrations of mononuclear inflammatory cells (A, B) and presumably extracellular, amastigote-like structures (C). Bursa fabricii of a nestling Alpine swift with depletion of the medullary follicle with lymphocytolysis (asterisk) and a distinct epithelium (arrows) (D)." figureDoi="http://doi.org/10.5281/zenodo.10933816" httpUri="https://zenodo.org/record/10933816/files/figure.png" pageId="3">Fig. 5C</figureCitation>
|
||
). No trypomastigotes could be identified within blood vessels.
|
||
</paragraph>
|
||
<paragraph id="365CE739FFEAFF95B258FD19974FFC76" blockId="3.[818,1488,148,976]" pageId="3">
|
||
The bursa fabricii examined in two birds demonstrated a depletion of the medullary follicles characterized by moderate to severe lymphocytolysis, surrounded by a distinct epithelium (
|
||
<figureCitation id="AED8FBBCFFEAFF95B5DEFCD1971AFCAA" box="[1239,1311,817,836]" captionStart="Fig" captionStartId="4.[100,130,766,783]" captionTargetBox="[207,1380,150,736]" captionTargetId="figure-311@4.[206,1381,148,738]" captionTargetPageId="4" captionText="Fig. 5. Skeletal musculature of a nestling Alpine swift showing infiltrations of mononuclear inflammatory cells (A, B) and presumably extracellular, amastigote-like structures (C). Bursa fabricii of a nestling Alpine swift with depletion of the medullary follicle with lymphocytolysis (asterisk) and a distinct epithelium (arrows) (D)." figureDoi="http://doi.org/10.5281/zenodo.10933816" httpUri="https://zenodo.org/record/10933816/files/figure.png" pageId="3">Fig. 5D</figureCitation>
|
||
). The spleens were difficult to assess due to autolytic changes. The lymphoid follicles were hypocellular and could not be clearly discriminated from the perivascular sheets, indicative of a mild lymphoid depletion.
|
||
</paragraph>
|
||
<paragraph id="365CE739FFEAFF95B258FC419654FC3E" blockId="3.[818,1488,148,976]" pageId="3">All other organs were generally unremarkable with no indications of any other underlying diseases.</paragraph>
|
||
<paragraph id="365CE739FFEAFF95B23BFC1C961EFBFE" blockId="3.[818,1051,1020,1040]" box="[818,1051,1020,1040]" pageId="3">
|
||
<heading id="6D145055FFEAFF95B23BFC1C961EFBFE" box="[818,1051,1020,1040]" fontSize="36" level="2" pageId="3" reason="3">
|
||
<emphasis id="04973B2BFFEAFF95B23BFC1C961EFBFE" bold="true" box="[818,1051,1020,1040]" italics="true" pageId="3">3.3. Ancillary diagnostics</emphasis>
|
||
</heading>
|
||
</paragraph>
|
||
<paragraph id="365CE739FFEAFF95B258FBD497A5FB8D" blockId="3.[818,1488,1076,1208]" pageId="3">All birds (n =5) tested negative for both West Nile and Usutu viruses. Both birds examined for Polyoma- and Circovirus tested negative.</paragraph>
|
||
<paragraph id="365CE739FFEAFF95B258FB8C91F3FB59" blockId="3.[818,1488,1076,1208]" pageId="3">A bacteriological examination of liver, lung, and spleen in one nestling resulted in growth of unspecific flora interpreted as post-mortem overgrowth.</paragraph>
|
||
<paragraph id="365CE739FFEAFF95B23BFB039626FB19" blockId="3.[818,1059,1251,1271]" box="[818,1059,1251,1271]" pageId="3">
|
||
<heading id="6D145055FFEAFF95B23BFB039626FB19" box="[818,1059,1251,1271]" fontSize="36" level="2" pageId="3" reason="3">
|
||
<emphasis id="04973B2BFFEAFF95B23BFB039626FB19" bold="true" box="[818,1059,1251,1271]" italics="true" pageId="3">3.4. Molecular diagnostics</emphasis>
|
||
</heading>
|
||
</paragraph>
|
||
<paragraph id="365CE739FFEAFF92B258FAFC92CDFC8A" blockId="3.[818,1488,1307,1439]" lastBlockId="4.[100,771,849,1175]" lastPageId="4" pageId="3">
|
||
The initial
|
||
<taxonomicName id="F1E39CBAFFEAFF95B2B7FAFB9657FAC1" box="[958,1106,1307,1327]" class="Kinetoplastea" family="Trypanosomatidae" genus="Leishmania" kingdom="Protozoa" order="Trypanosomatida" pageId="3" phylum="Euglenozoa" rank="species" species="undetermined">
|
||
<emphasis id="04973B2BFFEAFF95B2B7FAFB9627FAC1" bold="true" box="[958,1058,1307,1327]" italics="true" pageId="3">Leishmania</emphasis>
|
||
spp.
|
||
</taxonomicName>
|
||
PCR ITS1 resulted positive in all three samples with an approximate 350 bp length band, and thus larger than expected for
|
||
<taxonomicName id="F1E39CBAFFEAFF95B2BDFAB3964FFA89" box="[948,1098,1363,1383]" class="Kinetoplastea" family="Trypanosomatidae" genus="Leishmania" kingdom="Protozoa" order="Trypanosomatida" pageId="3" phylum="Euglenozoa" rank="species" species="undetermined">
|
||
<emphasis id="04973B2BFFEAFF95B2BDFAB3961DFA89" bold="true" box="[948,1048,1363,1383]" italics="true" pageId="3">Leishmania</emphasis>
|
||
spp.
|
||
</taxonomicName>
|
||
Sequencing analysis from one of these samples (consensus sequence of 260 bp) showed 97–100% identity with several
|
||
<taxonomicName id="F1E39CBAFFEAFF95B270FA6A961AFA71" box="[889,1055,1418,1439]" class="Kinetoplastea" family="Trypanosomatidae" genus="Trypanosoma" kingdom="Protozoa" order="Trypanosomatida" pageId="3" phylum="Euglenozoa" rank="species" species="undetermined">
|
||
<emphasis id="04973B2BFFEAFF95B270FA6A91F7FA70" bold="true" box="[889,1010,1418,1438]" italics="true" pageId="3">Trypanosoma</emphasis>
|
||
spp.
|
||
</taxonomicName>
|
||
18S rRNA sequences, but with a low coverage (14–16%).
|
||
</paragraph>
|
||
<caption id="629CB7B1FFEAFF95B16DF84C97D5F853" ID-DOI="http://doi.org/10.5281/zenodo.10933814" ID-Zenodo-Dep="10933814" box="[100,1488,1963,1981]" httpUri="https://zenodo.org/record/10933814/files/figure.png" pageId="3" startId="3.[100,130,1964,1981]" targetBox="[339,1247,1480,1935]" targetPageId="3" targetType="figure">
|
||
<paragraph id="365CE739FFEAFF95B16DF84C97D5F853" blockId="3.[100,1488,1963,1981]" box="[100,1488,1963,1981]" pageId="3">
|
||
<emphasis id="04973B2BFFEAFF95B16DF84C9298F853" bold="true" box="[100,157,1964,1981]" pageId="3">Fig. 4.</emphasis>
|
||
Distribution of louse flies on an Alpine swift nestling (A) compared with the distribution of bruising on post-mortem examination with plumage removed (B).
|
||
</paragraph>
|
||
</caption>
|
||
<caption id="629CB7B1FFEDFF92B16DFD1E97D5FCC6" ID-DOI="http://doi.org/10.5281/zenodo.10933816" ID-Zenodo-Dep="10933816" httpUri="https://zenodo.org/record/10933816/files/figure.png" pageId="4" startId="4.[100,130,766,783]" targetBox="[207,1380,150,736]" targetPageId="4" targetType="figure">
|
||
<paragraph id="365CE739FFEDFF92B16DFD1E97D5FCC6" blockId="4.[100,1488,764,808]" pageId="4">
|
||
<emphasis id="04973B2BFFEDFF92B16DFD1E9298FCE0" bold="true" box="[100,157,766,783]" pageId="4">Fig. 5.</emphasis>
|
||
Skeletal musculature of a nestling Alpine swift showing infiltrations of mononuclear inflammatory cells (A, B) and presumably extracellular, amastigote-like structures (C). Bursa fabricii of a nestling Alpine swift with depletion of the medullary follicle with lymphocytolysis (asterisk) and a distinct epithelium (arrows) (D).
|
||
</paragraph>
|
||
</caption>
|
||
<paragraph id="365CE739FFEDFF92B18DFC8D92CCFB79" blockId="4.[100,771,849,1175]" pageId="4">
|
||
Tissue pools of the nestlings (3/3) were positive for
|
||
<taxonomicName id="F1E39CBAFFEDFF92B356FC8C9106FC6F" box="[607,771,876,897]" class="Kinetoplastea" family="Trypanosomatidae" genus="Trypanosoma" kingdom="Protozoa" order="Trypanosomatida" pageId="4" phylum="Euglenozoa" rank="species" species="undetermined">
|
||
<emphasis id="04973B2BFFEDFF92B356FC8C90DDFC6E" bold="true" box="[607,728,876,896]" italics="true" pageId="4">Trypanosoma</emphasis>
|
||
spp.
|
||
</taxonomicName>
|
||
based upon results from the 18SrRNA PCR (gel not shown).The amplicon from one nestling was sequenced and the consensus sequence of 539 bp (primers trimmed GenBank accession no. OR598759) which showed an identity of 99.63% with 100% coverage with several sequences:
|
||
<taxonomicName id="F1E39CBAFFEDFF92B16DFC18930BFBE2" baseAuthorityName="Nandi and Bennett" baseAuthorityYear="1994" box="[100,270,1016,1036]" class="Kinetoplastea" family="Trypanosomatidae" genus="Trypanosoma" kingdom="Protozoa" order="Trypanosomatida" pageId="4" phylum="Euglenozoa" rank="species" species="corvi">
|
||
<emphasis id="04973B2BFFEDFF92B16DFC18930BFBE2" bold="true" box="[100,270,1016,1036]" italics="true" pageId="4">Trypanosoma corvi</emphasis>
|
||
</taxonomicName>
|
||
(JN006854 and AY461665, from
|
||
<taxonomicName id="F1E39CBAFFEDFF92B347FC189085FBE2" authorityName="Lacepede" authorityYear="1799" box="[590,640,1016,1036]" class="Aves" family="Accipitridae" genus="Buteo" kingdom="Animalia" order="Accipitriformes" pageId="4" phylum="Chordata" rank="genus">
|
||
<emphasis id="04973B2BFFEDFF92B347FC189085FBE2" bold="true" box="[590,640,1016,1036]" italics="true" pageId="4">Buteo</emphasis>
|
||
</taxonomicName>
|
||
from
|
||
<collectingCountry id="4EF4A7A9FFEDFF92B3B0FC189107FBE5" box="[697,770,1016,1035]" name="Czech Republic" pageId="4">Czechia</collectingCountry>
|
||
and
|
||
<taxonomicName id="F1E39CBAFFEDFF92B187FBF39320FBC9" authorityName="Linnaeus" authorityYear="1758" box="[142,293,1043,1063]" class="Aves" family="Corvidae" genus="Corvus" kingdom="Animalia" order="Passeriformes" pageId="4" phylum="Chordata" rank="species" species="frugilegus">
|
||
<emphasis id="04973B2BFFEDFF92B187FBF39320FBC9" bold="true" box="[142,293,1043,1063]" italics="true" pageId="4">Corvus frugilegus</emphasis>
|
||
</taxonomicName>
|
||
from the
|
||
<collectingCountry id="4EF4A7A9FFEDFF92B08DFBF49024FBC9" box="[388,545,1044,1063]" name="United Kingdom" pageId="4">United Kingdom</collectingCountry>
|
||
, respectively),
|
||
<taxonomicName id="F1E39CBAFFEDFF92B3B9FBF392B0FBAA" class="Kinetoplastea" family="Trypanosomatidae" genus="Trypanosoma" kingdom="Protozoa" order="Trypanosomatida" pageId="4" phylum="Euglenozoa" rank="species" species="undetermined">
|
||
<emphasis id="04973B2BFFEDFF92B3B9FBF39297FBAD" bold="true" italics="true" pageId="4">Trypanosoma</emphasis>
|
||
sp.
|
||
</taxonomicName>
|
||
(LZ 2011, JN006841, from
|
||
<taxonomicName id="F1E39CBAFFEDFF92B0CCFBCF9060FBAD" baseAuthorityName="Temminck" baseAuthorityYear="1815" box="[453,613,1071,1091]" class="Aves" family="Muscicapidae" genus="Ficedula" kingdom="Animalia" order="Passeriformes" pageId="4" phylum="Chordata" rank="species" species="albicollis">
|
||
<emphasis id="04973B2BFFEDFF92B0CCFBCF9060FBAD" bold="true" box="[453,613,1071,1091]" italics="true" pageId="4">Ficedula albicollis</emphasis>
|
||
</taxonomicName>
|
||
in
|
||
<collectingCountry id="4EF4A7A9FFEDFF92B38FFBD090D4FBAD" box="[646,721,1072,1091]" name="Czech Republic" pageId="4">Czechia</collectingCountry>
|
||
) and
|
||
<taxonomicName id="F1E39CBAFFEDFF92B16DFBAB92F8FB8E" box="[100,253,1099,1120]" class="Kinetoplastea" family="Trypanosomatidae" genus="Trypanosoma" kingdom="Protozoa" order="Trypanosomatida" pageId="4" phylum="Euglenozoa" rank="species" species="undetermined">
|
||
<emphasis id="04973B2BFFEDFF92B16DFBAB92D8FBB1" bold="true" box="[100,221,1099,1119]" italics="true" pageId="4">Trypanosoma</emphasis>
|
||
sp.
|
||
</taxonomicName>
|
||
AAT (AJ620557, from
|
||
<taxonomicName id="F1E39CBAFFEDFF92B0DEFBAB9047FB8E" box="[471,578,1099,1120]" class="Aves" family="Cracticidae" genus="Strepera" kingdom="Animalia" order="Passeriformes" pageId="4" phylum="Chordata" rank="species" species="undetermined">
|
||
<emphasis id="04973B2BFFEDFF92B0DEFBAB9025FBB1" bold="true" box="[471,544,1099,1119]" italics="true" pageId="4">Strepera</emphasis>
|
||
sp.
|
||
</taxonomicName>
|
||
, an Australian bird). Our sequence showed two mismatches with all the referenced sequences.
|
||
</paragraph>
|
||
<paragraph id="365CE739FFEDFF92B16DFB2E9382FB0C" blockId="4.[100,391,1230,1250]" box="[100,391,1230,1250]" pageId="4">
|
||
<heading id="6D145055FFEDFF92B16DFB2E9382FB0C" box="[100,391,1230,1250]" fontSize="36" level="2" pageId="4" reason="3">
|
||
<emphasis id="04973B2BFFEDFF92B16DFB2E9382FB0C" bold="true" box="[100,391,1230,1250]" italics="true" pageId="4">3.5. Evaluation of blood smears</emphasis>
|
||
</heading>
|
||
</paragraph>
|
||
<paragraph id="365CE739FFEDFF92B18DFAE69774FC3A" blockId="4.[132,770,1286,1306]" lastBlockId="4.[818,1491,849,1287]" pageId="4">Of the 72 evaluated blood smears from the 2022 season (colony B and C), 65 were from nestlings which fledged at the end of the season (90.2%). Overall, 27.8% of samples contained trypomastigotes (20/72). Seven animals died after 45 days-of-age, of which six (85.7%) had trypomastigotes and one did not (14.3%). Of the nestlings that fledged (n = 65), 14 had trypomastigotes (21.5%) visible on blood smear.</paragraph>
|
||
<paragraph id="365CE739FFEDFF92B258FC3D97CAFBB1" blockId="4.[818,1491,849,1287]" pageId="4">
|
||
Forty blood smears were examined in detail. An average trypomastigote burden of 7.9 trypomastigotes per 10 HPF was observed amongst positive animals (n = 20). In the sample with the highest burden, 56.4 trypomastigotes per 10 HPF were found. In most smears, the trypomastigotes were observed in clusters of rosette-like formations (
|
||
<figureCitation id="AED8FBBCFFEDFF92B485FBAC97C4FBB1" box="[1420,1473,1100,1119]" captionStart="Fig" captionStartId="4.[112,142,1964,1981]" captionTargetBox="[207,1380,1348,1934]" captionTargetId="figure-360@4.[206,1381,1346,1936]" captionTargetPageId="4" captionText="Fig. 6. Blood smears of nestling Alpine swifts with high (A) and moderate (B) trypomastigote burdens. Close-up of a trypomastigote between erythrocytes (C)." figureDoi="http://doi.org/10.5281/zenodo.10933818" httpUri="https://zenodo.org/record/10933818/files/figure.png" pageId="4">Fig. 6</figureCitation>
|
||
).
|
||
</paragraph>
|
||
<paragraph id="365CE739FFEDFF92B258FB8991A1FB5D" blockId="4.[818,1491,849,1287]" pageId="4">No clear difference was observed in the thrombocyte counts between positive and negative nestlings, with an average of 13.9 thrombocytes per 10 HPF.</paragraph>
|
||
<paragraph id="365CE739FFEDFF93B258FB5C90FEFF2D" blockId="4.[818,1491,849,1287]" lastBlockId="5.[100,770,148,195]" lastPageId="5" pageId="4">
|
||
Positive nestlings showed a higher level of mononuclear cells (18.6 per 10 HPF) in comparison with negative ones (8.2 per 10 HPF). Positive nestlings also showed a slightly higher level of granulocytes (3.9 per 10 HPF) when compared to negative ones (2.1 per 10 HPF). All counts, including standard deviation (SD), are compiled in
|
||
<tableCitation id="7B61D282FFECFF93B343FF509095FF2D" box="[586,656,176,195]" captionStart="Table 2" captionStartId="5.[100,150,1834,1850]" captionTargetPageId="5" captionText="Table 2 Average counts of 10 high-power-field counts (HPF) 400× magnification of positive and negative nestlings, with Standard deviation and range in brackets." httpUri="http://table.plazi.org/id/629CB7B1FFECFF93B16DF8CA978EF8BA" pageId="5" tableUuid="629CB7B1FFECFF93B16DF8CA978EF8BA">Table 2</tableCitation>
|
||
and
|
||
<figureCitation id="AED8FBBCFFECFF93B3C9FF5090FEFF2D" box="[704,763,176,195]" captionStart="Fig" captionStartId="6.[100,130,889,906]" captionTargetBox="[208,1379,148,861]" captionTargetId="graphics-782@6.[643,993,146,807]" captionTargetPageId="6" captionText="Fig. 7. Box plots comparing average counts per 10 HPF of granulocytes, mononuclear cells, and thrombocytes between positive (n =20) and negative (n =20) 45- day-old Alpine swift nestlings sampled in 2022." figureDoi="http://doi.org/10.5281/zenodo.10933820" httpUri="https://zenodo.org/record/10933820/files/figure.png" pageId="5">Fig. 7.</figureCitation>
|
||
</paragraph>
|
||
<caption id="629CB7B1FFEDFF92B179F84C97C0F853" ID-DOI="http://doi.org/10.5281/zenodo.10933818" ID-Zenodo-Dep="10933818" box="[112,1477,1963,1981]" httpUri="https://zenodo.org/record/10933818/files/figure.png" pageId="4" startId="4.[112,142,1964,1981]" targetBox="[207,1380,1348,1934]" targetPageId="4" targetType="figure">
|
||
<paragraph id="365CE739FFEDFF92B179F84C97C0F853" blockId="4.[112,1477,1963,1981]" box="[112,1477,1963,1981]" pageId="4">
|
||
<emphasis id="04973B2BFFEDFF92B179F84C92ACF853" bold="true" box="[112,169,1964,1981]" pageId="4">Fig. 6.</emphasis>
|
||
Blood smears of nestling Alpine swifts with high (A) and moderate (B) trypomastigote burdens. Close-up of a trypomastigote between erythrocytes (C).
|
||
</paragraph>
|
||
</caption>
|
||
<paragraph id="365CE739FFECFF93B16DFF0B93A3FF11" blockId="5.[100,422,235,255]" box="[100,422,235,255]" pageId="5">
|
||
<heading id="6D145055FFECFF93B16DFF0B93A3FF11" box="[100,422,235,255]" fontSize="36" level="2" pageId="5" reason="3">
|
||
<emphasis id="04973B2BFFECFF93B16DFF0B93A3FF11" bold="true" box="[100,422,235,255]" italics="true" pageId="5">3.6. Trypomastigote morphometrics</emphasis>
|
||
</heading>
|
||
</paragraph>
|
||
<paragraph id="365CE739FFECFF93B18DFEC49107FE81" blockId="5.[100,770,292,368]" pageId="5">
|
||
Trypomastigote morphometrics were similar in all evaluated blood smears (
|
||
<figureCitation id="AED8FBBCFFECFF93B1BBFEA092FDFEBD" box="[178,248,320,339]" captionStart="Fig" captionStartId="4.[112,142,1964,1981]" captionTargetBox="[207,1380,1348,1934]" captionTargetId="figure-360@4.[206,1381,1346,1936]" captionTargetPageId="4" captionText="Fig. 6. Blood smears of nestling Alpine swifts with high (A) and moderate (B) trypomastigote burdens. Close-up of a trypomastigote between erythrocytes (C)." figureDoi="http://doi.org/10.5281/zenodo.10933818" httpUri="https://zenodo.org/record/10933818/files/figure.png" pageId="5">Fig. 6C</figureCitation>
|
||
). The total length without the flagellum was on average 30.0 μm, based on n =36 from three blood smears, compiled in
|
||
<tableCitation id="7B61D282FFECFF93B3BEFEBB90F9FE81" box="[695,764,347,367]" captionStart="Table 3" captionStartId="6.[100,150,993,1009]" captionTargetPageId="6" captionText="Table 3 Average morphometric measurements of 36 trypomastigotes observed in 2022 from three 45-day-old Alpine swift nestlings. SD: Standard deviation." httpUri="http://table.plazi.org/id/629CB7B1FFEFFF90B16DFC0190B5FBCB" pageId="5" tableUuid="629CB7B1FFEFFF90B16DFC0190B5FBCB">Table 3</tableCitation>
|
||
.
|
||
</paragraph>
|
||
</subSubSection>
|
||
</treatment>
|
||
</document> |