286 lines
64 KiB
XML
286 lines
64 KiB
XML
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<mods:title id="D85604414C170B2FD130E068E40A1343">A Hotspot of Amoebae Diversity: 8 New Naked Amoebae Associated with the Planktonic Bloom-forming Cyanobacterium Microcystis</mods:title>
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<mods:namePart id="3DB1F9B731BFA3F5ADD3605C4D7F0CAE">Van Wichelen, Jeroen</mods:namePart>
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<mods:namePart id="65363304D4CA49A51621DF9EF6DC8E24">D’Hondt, Sofie</mods:namePart>
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<mods:namePart id="1986AE38309ED64CAAA6F1688C3A2CDF">Claeys, Myriam</mods:namePart>
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<mods:namePart id="624F1CB4455059B656FE0BE07F024B01">Vyverman, Wim</mods:namePart>
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<mods:namePart id="F3BAC2B741662D0206619642C9C58AD2">Berney, Cédric</mods:namePart>
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<mods:namePart id="3EE4E3DFB49BDB410937540B4D2366F5">Bass, David</mods:namePart>
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<mods:namePart id="A3493CC04B9F5F0DABADAD2245AD6FA5">Vanormelingen, Pieter</mods:namePart>
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<mods:title id="413B096194DB35247D4253015AC7ACF0">Acta Protozoologica</mods:title>
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<mods:date id="A2C98672DC8A5A4E23544941D18B15EB">2016</mods:date>
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<treatment id="0228D806FFA4FF8AFF7EFB66FE1F63D5" ID-DOI="http://doi.org/10.5281/zenodo.12538868" ID-GBIF-Taxon="233769608" ID-Zenodo-Dep="12538868" LSID="urn:lsid:plazi:treatment:0228D806FFA4FF8AFF7EFB66FE1F63D5" httpUri="http://treatment.plazi.org/id/0228D806FFA4FF8AFF7EFB66FE1F63D5" lastPageId="6" lastPageNumber="65" pageId="4" pageNumber="65">
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<subSubSection id="C29B3A9BFFA4FF88FF7EFB66FEEE64BC" box="[170,278,1186,1213]" pageId="4" pageNumber="65" type="nomenclature">
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<paragraph id="8A3E6910FFA4FF88FF7EFB66FEEE64BC" blockId="4.[132,779,1186,1945]" box="[170,278,1186,1213]" pageId="4" pageNumber="65">
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<heading id="D176DE7CFFA4FF88FF7EFB66FEEE64BC" box="[170,278,1186,1213]" fontSize="11" level="2" pageId="4" pageNumber="65" reason="5">
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<taxonomicName id="4D811293FFA4FF88FF7EFB66FEEE64BC" ID-CoL="85VM" authority="Bovee, 1965" box="[170,278,1186,1213]" class="Discosea" family="Vannellidae" genus="Vannella" kingdom="Protozoa" order="Vannellida" pageId="4" pageNumber="65" phylum="Amoebozoa" rank="genus">
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<emphasis id="B8F5B502FFA4FF88FF7EFB66FEEE64BC" bold="true" box="[170,278,1186,1213]" italics="true" pageId="4" pageNumber="65">Vannella</emphasis>
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</taxonomicName>
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</heading>
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</paragraph>
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</subSubSection>
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<subSubSection id="C29B3A9BFFA4FF8AFF7EFB08FE1F63D5" lastPageId="6" lastPageNumber="67" pageId="4" pageNumber="65" type="description">
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<paragraph id="8A3E6910FFA4FF88FF7EFB08FD4166A6" blockId="4.[132,779,1186,1945]" pageId="4" pageNumber="65">
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The vannellid strains belonged to two lineages in different parts of the
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<taxonomicName id="4D811293FFA4FF88FE4CFB2BFE076508" authority="Bovee, 1965" box="[408,511,1263,1289]" class="Discosea" family="Vannellidae" genus="Vannella" kingdom="Protozoa" order="Vannellida" pageId="4" pageNumber="65" phylum="Amoebozoa" rank="genus">
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<emphasis id="B8F5B502FFA4FF88FE4CFB2BFE076508" box="[408,511,1263,1289]" italics="true" pageId="4" pageNumber="65">Vannella</emphasis>
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</taxonomicName>
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ML phylogeny (
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<figureCitation id="12BA7595FFA4FF88FD0FFB2BFF52652A" captionStart="Fig" captionStartId="6.[132,167,1694,1716]" captionTargetId="figure-8@5.[180,1401,222,1933]" captionTargetPageId="5" captionText="Fig. 1.A – 18S rDNA maximum likelihood phylogeny of Vannella, including Microcystis–associated strains (in bold). ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of V. planctonica (strainsA2FBB: 2, 4, 5, 9–13 andA4P4ZHB:1, 3, 6–8) showing locomotive trophozoites (1–8), a grazing amoeba with a Microcystis cell inside a food vacuole (9), floating forms (10–11) and a cyst stage (12–13). C – LM pictures of V. simplex (strain A17WVB) showing the floating form (14), locomotive amoebae (15–18) with the presence of food vacuoles containing partly digested Microcystis cells (17), posteriorly adhered fecal pellets (15–16), a long flagellum-like pseudopodium encircling a Microcystis cell (17–18) and grazing amoebae on a colony of Microcystis aeruginosa with expanded (arrow) or contracted (arrowhead) flagellum-like pseudopodia visible in some of the trophozoites (19). The presence of a contracticle vacuole, a nucleus or a cyst opening is indicated by a black arrow, a black arrowhead or a white arrow respectively. Scale bars: 20 µm." figureDoi="http://doi.org/10.5281/zenodo.12538716" httpUri="https://zenodo.org/record/12538716/files/figure.png" pageId="4" pageNumber="65">Fig. 1A</figureCitation>
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). The first lineage had low bootstrap and moderate PP support and contained four
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<taxonomicName id="4D811293FFA4FF88FDD5FAF1FD71654F" authority="Lemmermann, 1907 " box="[513,649,1333,1358]" class="Cyanobacteriia" family="Microcystaceae" genus="Microcystis" kingdom="Bacteria" order="Cyanobacteriales" pageId="4" pageNumber="65" phylum="Cyanobacteria" rank="genus">
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<emphasis id="B8F5B502FFA4FF88FDD5FAF1FD71654F" box="[513,649,1333,1358]" italics="true" pageId="4" pageNumber="65">Microcystis</emphasis>
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</taxonomicName>
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-associated strains interspersed between 3
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<taxonomicName id="4D811293FFA4FF88FDD1FA92FD2B6571" authorityName="Bovee" authorityYear="1965" box="[517,723,1366,1392]" class="Discosea" family="Vannellidae" genus="Vannella" kingdom="Protozoa" order="Vannellida" pageId="4" pageNumber="65" phylum="Amoebozoa" rank="species" species="simplex">
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<emphasis id="B8F5B502FFA4FF88FDD1FA92FD2B6571" box="[517,723,1366,1392]" italics="true" pageId="4" pageNumber="65">Vannella simplex</emphasis>
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</taxonomicName>
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and 1 uncultured clone sequence (
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<emphasis id="B8F5B502FFA4FF88FDD4FABEFDF76592" box="[512,527,1402,1427]" italics="true" pageId="4" pageNumber="65">p</emphasis>
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-distances 0.3–3.5%, number of differing positions 4–63). Its sister taxon was
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<taxonomicName id="4D811293FFA4FF88FF69FA7AFE5C65D9" authorityName="Smirnov & Brown" authorityYear="2000" box="[189,420,1470,1496]" class="Discosea" family="Vannellidae" genus="Vannella" kingdom="Protozoa" order="Vannellida" pageId="4" pageNumber="65" phylum="Amoebozoa" rank="species" species="persistens">
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<emphasis id="B8F5B502FFA4FF88FF69FA7AFE5C65D9" box="[189,420,1470,1496]" italics="true" pageId="4" pageNumber="65">Vannella persistens</emphasis>
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</taxonomicName>
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(2.0–3.6%, 24–68). The second lineage (upper part of the phylogeny) had moderate bootstrap and maximal PP support and consisted of six
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<taxonomicName id="4D811293FFA4FF88FF79F9E2FECD663E" authority="Lemmermann, 1907 " box="[173,309,1574,1599]" class="Cyanobacteriia" family="Microcystaceae" genus="Microcystis" kingdom="Bacteria" order="Cyanobacteriales" pageId="4" pageNumber="65" phylum="Cyanobacteria" rank="genus">
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<emphasis id="B8F5B502FFA4FF88FF79F9E2FECD663E" box="[173,309,1574,1599]" italics="true" pageId="4" pageNumber="65">Microcystis</emphasis>
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</taxonomicName>
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-associated
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<taxonomicName id="4D811293FFA4FF88FE69F9E1FDDC663E" authority="Bovee, 1965" box="[445,548,1573,1599]" class="Discosea" family="Vannellidae" genus="Vannella" kingdom="Protozoa" order="Vannellida" pageId="4" pageNumber="65" phylum="Amoebozoa" rank="genus">
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<emphasis id="B8F5B502FFA4FF88FE69F9E1FDDC663E" box="[445,548,1573,1599]" italics="true" pageId="4" pageNumber="65">Vannella</emphasis>
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</taxonomicName>
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strains (
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<emphasis id="B8F5B502FFA4FF88FD53F9E2FD6E663E" box="[647,662,1574,1599]" italics="true" pageId="4" pageNumber="65">p</emphasis>
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-distances 0.8–3.0%, 13–28 differing positions). Its closest relatives were unidentified
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<taxonomicName id="4D811293FFA4FF88FE75F9AEFDF06685" authority="Bovee, 1965" box="[417,520,1642,1668]" class="Discosea" family="Vannellidae" genus="Vannella" kingdom="Protozoa" order="Vannellida" pageId="4" pageNumber="65" phylum="Amoebozoa" rank="genus">
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<emphasis id="B8F5B502FFA4FF88FE75F9AEFDF06685" box="[417,520,1642,1668]" italics="true" pageId="4" pageNumber="65">Vannella</emphasis>
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</taxonomicName>
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strains or uncultured clones from natural samples (2.4–9.1%, 32–98).
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</paragraph>
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<paragraph id="8A3E6910FFA4FF88FF7EF96BFBE46296" blockId="4.[132,779,1186,1945]" lastBlockId="4.[816,1463,223,1941]" pageId="4" pageNumber="65">
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The locomotive cells of the vannellid strains A17
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<collectionCode id="EC90F1D5FFA4FF88FF62F916FF0666ED" box="[182,254,1746,1772]" pageId="4" pageNumber="65">WVB</collectionCode>
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, A1
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<collectionCode id="EC90F1D5FFA4FF88FEFAF916FE8966ED" box="[302,369,1746,1772]" country="Slovakia" httpUri="http://grbio.org/cool/8idg-801q" name="Šarišské Múzeum" pageId="4" pageNumber="65" type="Herbarium">SMB</collectionCode>
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, A6
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<collectionCode id="EC90F1D5FFA4FF88FE77F916FE2666ED" box="[419,478,1746,1772]" country="United Kingdom" httpUri="http://biocol.org/urn:lsid:biocol.org:col:13204" lsid="urn:lsid:biocol.org:col:13204" name="Hebden Bridge Literary and Scientific Society" pageId="4" pageNumber="65" type="Herbarium">HEB</collectionCode>
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and A7
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<collectionCode id="EC90F1D5FFA4FF88FD96F916FD8766ED" box="[578,639,1746,1772]" country="United Kingdom" httpUri="http://biocol.org/urn:lsid:biocol.org:col:13204" lsid="urn:lsid:biocol.org:col:13204" name="Hebden Bridge Literary and Scientific Society" pageId="4" pageNumber="65" type="Herbarium">HEB</collectionCode>
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, all closely related to
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<taxonomicName id="4D811293FFA4FF88FF28F931FE8D670F" authorityName="Bovee" authorityYear="1965" box="[252,373,1780,1806]" class="Discosea" family="Vannellidae" genus="Vannella" kingdom="Protozoa" order="Vannellida" pageId="4" pageNumber="65" phylum="Amoebozoa" rank="species" species="simplex">
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<emphasis id="B8F5B502FFA4FF88FF28F931FE8D670F" box="[252,373,1780,1806]" italics="true" pageId="4" pageNumber="65">V. simplex</emphasis>
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</taxonomicName>
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(
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<figureCitation id="12BA7595FFA4FF88FE52F930FE19670E" box="[390,481,1780,1807]" captionStart="Fig" captionStartId="6.[132,167,1694,1716]" captionTargetId="figure-8@5.[180,1401,222,1933]" captionTargetPageId="5" captionText="Fig. 1.A – 18S rDNA maximum likelihood phylogeny of Vannella, including Microcystis–associated strains (in bold). ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of V. planctonica (strainsA2FBB: 2, 4, 5, 9–13 andA4P4ZHB:1, 3, 6–8) showing locomotive trophozoites (1–8), a grazing amoeba with a Microcystis cell inside a food vacuole (9), floating forms (10–11) and a cyst stage (12–13). C – LM pictures of V. simplex (strain A17WVB) showing the floating form (14), locomotive amoebae (15–18) with the presence of food vacuoles containing partly digested Microcystis cells (17), posteriorly adhered fecal pellets (15–16), a long flagellum-like pseudopodium encircling a Microcystis cell (17–18) and grazing amoebae on a colony of Microcystis aeruginosa with expanded (arrow) or contracted (arrowhead) flagellum-like pseudopodia visible in some of the trophozoites (19). The presence of a contracticle vacuole, a nucleus or a cyst opening is indicated by a black arrow, a black arrowhead or a white arrow respectively. Scale bars: 20 µm." figureDoi="http://doi.org/10.5281/zenodo.12538716" httpUri="https://zenodo.org/record/12538716/files/figure.png" pageId="4" pageNumber="65">Fig. 1C</figureCitation>
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), shared a mainly flabellate to semicircular, rarely spatulate shape, a length/ width ratio of 0.4–1.6 (mean 0.8) and an average coverage with hyaloplasm about half of the cell length anteriorly, but differed in average size. Locomotive forms of strains A17
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<collectionCode id="EC90F1D5FFA4FF88FC6CFF1BFBF860F8" box="[952,1024,223,249]" pageId="4" pageNumber="65">WVB</collectionCode>
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, A1
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<collectionCode id="EC90F1D5FFA4FF88FBFAFF1BFB9760F8" box="[1070,1135,223,249]" country="Slovakia" httpUri="http://grbio.org/cool/8idg-801q" name="Šarišské Múzeum" pageId="4" pageNumber="65" type="Herbarium">SMB</collectionCode>
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and A6
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<collectionCode id="EC90F1D5FFA4FF88FB05FF1BFAF460F8" box="[1233,1292,223,249]" country="United Kingdom" httpUri="http://biocol.org/urn:lsid:biocol.org:col:13204" lsid="urn:lsid:biocol.org:col:13204" name="Hebden Bridge Literary and Scientific Society" pageId="4" pageNumber="65" type="Herbarium">HEB</collectionCode>
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/A7
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<collectionCode id="EC90F1D5FFA4FF88FAECFF1BFA8B60F8" box="[1336,1395,223,249]" country="United Kingdom" httpUri="http://biocol.org/urn:lsid:biocol.org:col:13204" lsid="urn:lsid:biocol.org:col:13204" name="Hebden Bridge Literary and Scientific Society" pageId="4" pageNumber="65" type="Herbarium">HEB</collectionCode>
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were respectively 38–79 (mean 49), 20–56 (35) and 15–60 (31) µm long and respectively 45–83 (65), 20–61 (43) and 20–85 (41) µm wide. On several occasions, trophozoites of strain A17
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<collectionCode id="EC90F1D5FFA4FF88FB9FFEADFB6A6182" box="[1099,1170,361,387]" pageId="4" pageNumber="65">WVB</collectionCode>
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were observed containing a flagellum-like pseudopodium, either very long and straight or contracted and helical in appearance (
|
||
<figureCitation id="12BA7595FFA4FF88FCEDFE15FC6C61EA" box="[825,916,465,491]" captionStart="Fig" captionStartId="6.[132,167,1694,1716]" captionTargetId="figure-8@5.[180,1401,222,1933]" captionTargetPageId="5" captionText="Fig. 1.A – 18S rDNA maximum likelihood phylogeny of Vannella, including Microcystis–associated strains (in bold). ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of V. planctonica (strainsA2FBB: 2, 4, 5, 9–13 andA4P4ZHB:1, 3, 6–8) showing locomotive trophozoites (1–8), a grazing amoeba with a Microcystis cell inside a food vacuole (9), floating forms (10–11) and a cyst stage (12–13). C – LM pictures of V. simplex (strain A17WVB) showing the floating form (14), locomotive amoebae (15–18) with the presence of food vacuoles containing partly digested Microcystis cells (17), posteriorly adhered fecal pellets (15–16), a long flagellum-like pseudopodium encircling a Microcystis cell (17–18) and grazing amoebae on a colony of Microcystis aeruginosa with expanded (arrow) or contracted (arrowhead) flagellum-like pseudopodia visible in some of the trophozoites (19). The presence of a contracticle vacuole, a nucleus or a cyst opening is indicated by a black arrow, a black arrowhead or a white arrow respectively. Scale bars: 20 µm." figureDoi="http://doi.org/10.5281/zenodo.12538716" httpUri="https://zenodo.org/record/12538716/files/figure.png" pageId="4" pageNumber="65">Fig. 1C</figureCitation>
|
||
17–19). The trophozoites of strain A17
|
||
<collectionCode id="EC90F1D5FFA4FF88FAA4FE15FA4F61EA" box="[1392,1463,465,491]" pageId="4" pageNumber="65">WVB</collectionCode>
|
||
were always covered with fecal pellets. For the other 3 strains this feature disappeared after several months of cultivation, however fecal pellets of about 9 µm in diameter could always be observed freely dispersed in the culture medium.
|
||
</paragraph>
|
||
<paragraph id="8A3E6910FFA4FF88FC82FD64FB826751" blockId="4.[816,1463,223,1941]" pageId="4" pageNumber="65">
|
||
Locomotive amoebae of strains A4P4
|
||
<collectionCode id="EC90F1D5FFA4FF88FADDFD64FABE62BB" box="[1289,1350,672,698]" pageId="4" pageNumber="65">ZHB</collectionCode>
|
||
, A2
|
||
<collectionCode id="EC90F1D5FFA4FF88FAA6FD64FA4A62BB" box="[1394,1458,672,698]" country="Slovakia" httpUri="http://grbio.org/cool/8idg-801q" name="Šarišské Múzeum" pageId="4" pageNumber="65" type="Herbarium">SMB</collectionCode>
|
||
, A2
|
||
<collectionCode id="EC90F1D5FFA4FF88FC81FD06FC7662DD" box="[853,910,706,732]" pageId="4" pageNumber="65">FBB</collectionCode>
|
||
, A1
|
||
<collectionCode id="EC90F1D5FFA4FF88FC68FD06FBDF62DD" box="[956,1063,706,732]" pageId="4" pageNumber="65">DVDPB</collectionCode>
|
||
, A2
|
||
<collectionCode id="EC90F1D5FFA4FF88FB8DFD06FB3862DD" box="[1113,1216,706,732]" pageId="4" pageNumber="65">DVDPB</collectionCode>
|
||
, A3
|
||
<collectionCode id="EC90F1D5FFA4FF88FB24FD06FAA062DD" box="[1264,1368,706,732]" pageId="4" pageNumber="65">DVDPB</collectionCode>
|
||
, all belonging to the second vannellid lineage, were fan-shaped however with considerable variations. They could adopt a semicircular (
|
||
<figureCitation id="12BA7595FFA4FF88FB5BFCEEFAD76345" box="[1167,1327,810,836]" captionStart-0="Fig" captionStart-1="Fig" captionStartId-0="6.[132,167,1694,1716]" captionStartId-1="7.[125,160,1721,1743]" captionTargetId-0="figure-8@5.[180,1401,222,1933]" captionTargetId-1="figure-9@8.[142,1454,223,1930]" captionTargetPageId-0="5" captionTargetPageId-1="8" captionText-0="Fig. 1.A – 18S rDNA maximum likelihood phylogeny of Vannella, including Microcystis–associated strains (in bold). ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of V. planctonica (strainsA2FBB: 2, 4, 5, 9–13 andA4P4ZHB:1, 3, 6–8) showing locomotive trophozoites (1–8), a grazing amoeba with a Microcystis cell inside a food vacuole (9), floating forms (10–11) and a cyst stage (12–13). C – LM pictures of V. simplex (strain A17WVB) showing the floating form (14), locomotive amoebae (15–18) with the presence of food vacuoles containing partly digested Microcystis cells (17), posteriorly adhered fecal pellets (15–16), a long flagellum-like pseudopodium encircling a Microcystis cell (17–18) and grazing amoebae on a colony of Microcystis aeruginosa with expanded (arrow) or contracted (arrowhead) flagellum-like pseudopodia visible in some of the trophozoites (19). The presence of a contracticle vacuole, a nucleus or a cyst opening is indicated by a black arrow, a black arrowhead or a white arrow respectively. Scale bars: 20 µm." captionText-1="Fig. 2. A – 18S rDNA maximum likelihood phylogeny of Korotnevella, including Microcystis-associated strains (in bold). ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of K. jeppesenii (strain A1JEPDK) showing trophozoites in non-oriented movement (1–5), during locomotion (6, 8) and during grazing on Microcystis aeruginosa cells (7) with the arrow indicating a lacuna-containing nucleolus. C – LM pictures of K. pelagolacustris (strains A8WVB (7, 12), A16WVB (6, 8, 11), A21WVB (1, 13), A54WVB (3–5) and A1LMS (2, 9, 10)) showing trophozoites in different stages of non-oriented movement (1–4, 6), the locomotive form (8–12), grazing amoebae with Microcystis cells inside food vacuoles (5, 10), the cyst stage (7) and the floating form (13).Arrows indicate the nucleus with clearly visible nucleolus (in 8, 10, 12). Scale bars: 20 µm." figureDoi-0="http://doi.org/10.5281/zenodo.12538716" figureDoi-1="http://doi.org/10.5281/zenodo.12538718" httpUri-0="https://zenodo.org/record/12538716/files/figure.png" httpUri-1="https://zenodo.org/record/12538718/files/figure.png" pageId="4" pageNumber="65">Figs 1B 1–2</figureCitation>
|
||
), flabellate (
|
||
<figureCitation id="12BA7595FFA4FF88FCEDFC88FC666366" box="[825,926,844,871]" captionStart="Fig" captionStartId="6.[132,167,1694,1716]" captionTargetId="figure-8@5.[180,1401,222,1933]" captionTargetPageId="5" captionText="Fig. 1.A – 18S rDNA maximum likelihood phylogeny of Vannella, including Microcystis–associated strains (in bold). ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of V. planctonica (strainsA2FBB: 2, 4, 5, 9–13 andA4P4ZHB:1, 3, 6–8) showing locomotive trophozoites (1–8), a grazing amoeba with a Microcystis cell inside a food vacuole (9), floating forms (10–11) and a cyst stage (12–13). C – LM pictures of V. simplex (strain A17WVB) showing the floating form (14), locomotive amoebae (15–18) with the presence of food vacuoles containing partly digested Microcystis cells (17), posteriorly adhered fecal pellets (15–16), a long flagellum-like pseudopodium encircling a Microcystis cell (17–18) and grazing amoebae on a colony of Microcystis aeruginosa with expanded (arrow) or contracted (arrowhead) flagellum-like pseudopodia visible in some of the trophozoites (19). The presence of a contracticle vacuole, a nucleus or a cyst opening is indicated by a black arrow, a black arrowhead or a white arrow respectively. Scale bars: 20 µm." figureDoi="http://doi.org/10.5281/zenodo.12538716" httpUri="https://zenodo.org/record/12538716/files/figure.png" pageId="4" pageNumber="65">Figs 1B</figureCitation>
|
||
<figureCitation id="12BA7595FFA4FF88FC78FC88FC216366" box="[940,985,844,871]" captionStart-0="Fig" captionStart-1="Fig" captionStartId-0="9.[125,160,1695,1717]" captionStartId-1="12.[132,167,1863,1885]" captionTargetBox-1="[132,1457,1336,1835]" captionTargetId-0="figure-10@10.[177,1411,223,1939]" captionTargetId-1="figure-482@12.[132,1457,1336,1835]" captionTargetPageId-0="10" captionTargetPageId-1="12" captionText-0="Fig. 4. A – 18S rDNA maximum likelihood phylogeny of the family Hartmannellidae, including Microcystis-associated strains of Copromyxa (in bold). ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of C. microcystidis showing locomotive trophozoites (1–7, 10) containing a pronounced hyaline cap (white arrowheads), a vesicular nucleus with globular nucleolus (white arrows), big crystals (black arrows) and Microcystis cells inside food vacuoles (black arrowheads), non-oriented moving trophozoites (8–9, 11), a grazing amoeba capturing a Microcystis cell (12–14), the floating form (15) and a double-walled cyst stage (16). C – LM pictures of C. vandevyveri showing locomotive trophozoites (1, 6–9), trophozoites during non-oriented movement (2–5, 10–12) and floating forms (13–15). Black arrows indicate the presence of small, refractive crystals in the cytoplasm, the white arrow shows the vesicular nucleus with a globular lacuna-containing nucleolus, white arrowheads indicate a pronounced hyaline cap. Ingested Microcystis cells are visible in (9) and (11). Scale bars: 20 µm." captionText-1="Fig. 5. LM pictures of Schoutedamoeba minuta showing trophozoites in non-oriented movement (a–f) and the limax-shaped locomotive form (g–i). The presence of a pronounced hyaline cap, small adhesive uroidal filaments and tiny granules in the cytoplasm are indicated with white arrowheads, black arrows and a white arrow respectively. Scale bars: 20 µm." figureDoi-0="http://doi.org/10.5281/zenodo.12538723" figureDoi-1="http://doi.org/10.5281/zenodo.12538725" httpUri-0="https://zenodo.org/record/12538723/files/figure.png" httpUri-1="https://zenodo.org/record/12538725/files/figure.png" pageId="4" pageNumber="65">4–5</figureCitation>
|
||
) or spatulate shape containing a pronounced tail up to 21 µm long (
|
||
<figureCitation id="12BA7595FFA4FF88FB79FCABFAF06388" box="[1197,1288,879,905]" captionStart="Fig" captionStartId="6.[132,167,1694,1716]" captionTargetId="figure-8@5.[180,1401,222,1933]" captionTargetPageId="5" captionText="Fig. 1.A – 18S rDNA maximum likelihood phylogeny of Vannella, including Microcystis–associated strains (in bold). ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of V. planctonica (strainsA2FBB: 2, 4, 5, 9–13 andA4P4ZHB:1, 3, 6–8) showing locomotive trophozoites (1–8), a grazing amoeba with a Microcystis cell inside a food vacuole (9), floating forms (10–11) and a cyst stage (12–13). C – LM pictures of V. simplex (strain A17WVB) showing the floating form (14), locomotive amoebae (15–18) with the presence of food vacuoles containing partly digested Microcystis cells (17), posteriorly adhered fecal pellets (15–16), a long flagellum-like pseudopodium encircling a Microcystis cell (17–18) and grazing amoebae on a colony of Microcystis aeruginosa with expanded (arrow) or contracted (arrowhead) flagellum-like pseudopodia visible in some of the trophozoites (19). The presence of a contracticle vacuole, a nucleus or a cyst opening is indicated by a black arrow, a black arrowhead or a white arrow respectively. Scale bars: 20 µm." figureDoi="http://doi.org/10.5281/zenodo.12538716" httpUri="https://zenodo.org/record/12538716/files/figure.png" pageId="4" pageNumber="65">Fig. 1B</figureCitation>
|
||
<figureCitation id="12BA7595FFA4FF88FAC5FCABFAD96388" box="[1297,1313,879,905]" captionStart="Fig" captionStartId="9.[127,162,1135,1157]" captionTargetBox="[127,1452,221,1108]" captionTargetId="figure-7@9.[127,1452,221,1108]" captionTargetPageId="9" captionText="Fig. 3. TEM pictures of the scales on the cell surface of Korotnevella jeppesenii (a, c) and K. pelagolacustris (b, d) trophozoites viewed from top (a, b) and side (c, d). Scale bars: 500 nm." figureDoi="http://doi.org/10.5281/zenodo.12538720" httpUri="https://zenodo.org/record/12538720/files/figure.png" pageId="4" pageNumber="65">3</figureCitation>
|
||
,
|
||
<figureCitation id="12BA7595FFA4FF88FAE4FCABFAB86388" box="[1328,1344,879,905]" captionStart="Fig" captionStartId="17.[125,160,1778,1800]" captionTargetId="figure-10@18.[148,1448,222,1933]" captionTargetPageId="18" captionText="Fig. 8. A – 18S rDNA maximum likelihood phylogeny of Angulamoeba. Sequences new for this study are indicated in bold. ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of Angulamoeba microcystivorans (strains A1WVB (1, 2, 4) and A4WVB (3)) showing the characteristic rusty-colored plasmodium-like mucilage matrix with embedded trophozoites devouring a culture of Microcystis aeruginosa colonies (1–2), multi-vacuolated trophozoites that are seemingly connected by their filopodia (3) and amoeba-flagellates in different stages of transition (4). Scale bars: 200 µm (1), 100 µm (2), 20 µm (3, 4)." figureDoi="http://doi.org/10.5281/zenodo.12538733" httpUri="https://zenodo.org/record/12538733/files/figure.png" pageId="4" pageNumber="65">8</figureCitation>
|
||
). Feeding amoeba attached on
|
||
<taxonomicName id="4D811293FFA4FF88FBF9FC56FB4D63AA" box="[1069,1205,914,939]" class="Cyanobacteriia" family="Microcystaceae" genus="Microcystis" kingdom="Bacteria" order="Cyanobacteriales" pageId="4" pageNumber="65" phylum="Cyanobacteria" rank="genus">
|
||
<emphasis id="B8F5B502FFA4FF88FBF9FC56FB4D63AA" box="[1069,1205,914,939]" italics="true" pageId="4" pageNumber="65">Microcystis</emphasis>
|
||
</taxonomicName>
|
||
colonies (
|
||
<figureCitation id="12BA7595FFA4FF88FAE2FC55FA6B63AD" box="[1334,1427,913,940]" captionStart="Fig" captionStartId="6.[132,167,1694,1716]" captionTargetId="figure-8@5.[180,1401,222,1933]" captionTargetPageId="5" captionText="Fig. 1.A – 18S rDNA maximum likelihood phylogeny of Vannella, including Microcystis–associated strains (in bold). ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of V. planctonica (strainsA2FBB: 2, 4, 5, 9–13 andA4P4ZHB:1, 3, 6–8) showing locomotive trophozoites (1–8), a grazing amoeba with a Microcystis cell inside a food vacuole (9), floating forms (10–11) and a cyst stage (12–13). C – LM pictures of V. simplex (strain A17WVB) showing the floating form (14), locomotive amoebae (15–18) with the presence of food vacuoles containing partly digested Microcystis cells (17), posteriorly adhered fecal pellets (15–16), a long flagellum-like pseudopodium encircling a Microcystis cell (17–18) and grazing amoebae on a colony of Microcystis aeruginosa with expanded (arrow) or contracted (arrowhead) flagellum-like pseudopodia visible in some of the trophozoites (19). The presence of a contracticle vacuole, a nucleus or a cyst opening is indicated by a black arrow, a black arrowhead or a white arrow respectively. Scale bars: 20 µm." figureDoi="http://doi.org/10.5281/zenodo.12538716" httpUri="https://zenodo.org/record/12538716/files/figure.png" pageId="4" pageNumber="65">Fig. 1B</figureCitation>
|
||
9) had a greatest linear dimension of 15–50 µm (mean 31). The hyaloplasm formed a broad margin around the cell and covered 13–71% (40%) of the cell anteriorly. When floating cells settled and became locomotive, the frontal hyaline area could display a wavy front edge (
|
||
<figureCitation id="12BA7595FFA4FF88FCEDFBA5FC6E647A" box="[825,918,1121,1147]" captionStart="Fig" captionStartId="6.[132,167,1694,1716]" captionTargetId="figure-8@5.[180,1401,222,1933]" captionTargetPageId="5" captionText="Fig. 1.A – 18S rDNA maximum likelihood phylogeny of Vannella, including Microcystis–associated strains (in bold). ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of V. planctonica (strainsA2FBB: 2, 4, 5, 9–13 andA4P4ZHB:1, 3, 6–8) showing locomotive trophozoites (1–8), a grazing amoeba with a Microcystis cell inside a food vacuole (9), floating forms (10–11) and a cyst stage (12–13). C – LM pictures of V. simplex (strain A17WVB) showing the floating form (14), locomotive amoebae (15–18) with the presence of food vacuoles containing partly digested Microcystis cells (17), posteriorly adhered fecal pellets (15–16), a long flagellum-like pseudopodium encircling a Microcystis cell (17–18) and grazing amoebae on a colony of Microcystis aeruginosa with expanded (arrow) or contracted (arrowhead) flagellum-like pseudopodia visible in some of the trophozoites (19). The presence of a contracticle vacuole, a nucleus or a cyst opening is indicated by a black arrow, a black arrowhead or a white arrow respectively. Scale bars: 20 µm." figureDoi="http://doi.org/10.5281/zenodo.12538716" httpUri="https://zenodo.org/record/12538716/files/figure.png" pageId="4" pageNumber="65">Figs 1B</figureCitation>
|
||
<figureCitation id="12BA7595FFA4FF88FC4AFBA5FC57647A" box="[926,943,1121,1147]" captionStart="Fig" captionStartId="7.[125,160,1721,1743]" captionTargetId="figure-9@8.[142,1454,223,1930]" captionTargetPageId="8" captionText="Fig. 2. A – 18S rDNA maximum likelihood phylogeny of Korotnevella, including Microcystis-associated strains (in bold). ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of K. jeppesenii (strain A1JEPDK) showing trophozoites in non-oriented movement (1–5), during locomotion (6, 8) and during grazing on Microcystis aeruginosa cells (7) with the arrow indicating a lacuna-containing nucleolus. C – LM pictures of K. pelagolacustris (strains A8WVB (7, 12), A16WVB (6, 8, 11), A21WVB (1, 13), A54WVB (3–5) and A1LMS (2, 9, 10)) showing trophozoites in different stages of non-oriented movement (1–4, 6), the locomotive form (8–12), grazing amoebae with Microcystis cells inside food vacuoles (5, 10), the cyst stage (7) and the floating form (13).Arrows indicate the nucleus with clearly visible nucleolus (in 8, 10, 12). Scale bars: 20 µm." figureDoi="http://doi.org/10.5281/zenodo.12538718" httpUri="https://zenodo.org/record/12538718/files/figure.png" pageId="4" pageNumber="65">2</figureCitation>
|
||
,
|
||
<figureCitation id="12BA7595FFA4FF88FC6FFBA5FC34647A" box="[955,972,1121,1147]" captionStart="Fig" captionStartId="9.[125,160,1695,1717]" captionTargetId="figure-10@10.[177,1411,223,1939]" captionTargetPageId="10" captionText="Fig. 4. A – 18S rDNA maximum likelihood phylogeny of the family Hartmannellidae, including Microcystis-associated strains of Copromyxa (in bold). ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of C. microcystidis showing locomotive trophozoites (1–7, 10) containing a pronounced hyaline cap (white arrowheads), a vesicular nucleus with globular nucleolus (white arrows), big crystals (black arrows) and Microcystis cells inside food vacuoles (black arrowheads), non-oriented moving trophozoites (8–9, 11), a grazing amoeba capturing a Microcystis cell (12–14), the floating form (15) and a double-walled cyst stage (16). C – LM pictures of C. vandevyveri showing locomotive trophozoites (1, 6–9), trophozoites during non-oriented movement (2–5, 10–12) and floating forms (13–15). Black arrows indicate the presence of small, refractive crystals in the cytoplasm, the white arrow shows the vesicular nucleus with a globular lacuna-containing nucleolus, white arrowheads indicate a pronounced hyaline cap. Ingested Microcystis cells are visible in (9) and (11). Scale bars: 20 µm." figureDoi="http://doi.org/10.5281/zenodo.12538723" httpUri="https://zenodo.org/record/12538723/files/figure.png" pageId="4" pageNumber="65">4</figureCitation>
|
||
,
|
||
<figureCitation id="12BA7595FFA4FF88FC0DFBA5FC10647D" box="[985,1000,1121,1148]" captionStart="Fig" captionStartId="13.[125,160,1833,1855]" captionTargetId="figure-9@14.[135,1460,257,1939]" captionTargetPageId="14" captionText="Fig. 6. A – 18S rDNA maximum likelihood phylogeny of Vexillifera, including the Microcystis-associated strain (in bold). ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of locomotive (1–7, 12–13) and floating (8–11) amoebae. Microcystis cells inside food vacuoles are visible in (1–2) and (13). The arrow in (5) points to a temporal, uroid-like structure sometimes visible during locomotion. Scale bars: 20 µm." figureDoi="http://doi.org/10.5281/zenodo.12538727" httpUri="https://zenodo.org/record/12538727/files/figure.png" pageId="4" pageNumber="65">6</figureCitation>
|
||
) and sometimes contained longitudinal ridges (
|
||
<figureCitation id="12BA7595FFA4FF88FC58FB47FC10649C" box="[908,1000,1155,1181]" captionStart="Fig" captionStartId="6.[132,167,1694,1716]" captionTargetId="figure-8@5.[180,1401,222,1933]" captionTargetPageId="5" captionText="Fig. 1.A – 18S rDNA maximum likelihood phylogeny of Vannella, including Microcystis–associated strains (in bold). ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of V. planctonica (strainsA2FBB: 2, 4, 5, 9–13 andA4P4ZHB:1, 3, 6–8) showing locomotive trophozoites (1–8), a grazing amoeba with a Microcystis cell inside a food vacuole (9), floating forms (10–11) and a cyst stage (12–13). C – LM pictures of V. simplex (strain A17WVB) showing the floating form (14), locomotive amoebae (15–18) with the presence of food vacuoles containing partly digested Microcystis cells (17), posteriorly adhered fecal pellets (15–16), a long flagellum-like pseudopodium encircling a Microcystis cell (17–18) and grazing amoebae on a colony of Microcystis aeruginosa with expanded (arrow) or contracted (arrowhead) flagellum-like pseudopodia visible in some of the trophozoites (19). The presence of a contracticle vacuole, a nucleus or a cyst opening is indicated by a black arrow, a black arrowhead or a white arrow respectively. Scale bars: 20 µm." figureDoi="http://doi.org/10.5281/zenodo.12538716" httpUri="https://zenodo.org/record/12538716/files/figure.png" pageId="4" pageNumber="65">Fig. 1B</figureCitation>
|
||
<figureCitation id="12BA7595FFA4FF88FC27FB40FBFA649F" box="[1011,1026,1156,1182]" captionStart="Fig" captionStartId="15.[124,159,1777,1799]" captionTargetId="figure-9@16.[190,1408,222,1933]" captionTargetPageId="16" captionText="Fig. 7. A – 18S rDNA maximum likelihood phylogeny of Cochliopodium, including the Microcystis-associated strain (in bold). ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of locomotive amoebae clearly showing the surrounding hyaline sheet (black arrows) punctuated with microscales (best visible in 8), a few small subpseudopodia (black arrowheads) and trailing adhesive uroidal filaments (white arrows) (1–5), a tropho- zoite during and just after grazing on Microcystis cells showing a prominent fringe of folded hyaloplasm (white arrowheads) (6–9), the bell-shaped form on colonies of Microcystis aeruginosa (10–11) and floating amoebae (12–13). Scale bars: 20 µm." figureDoi="http://doi.org/10.5281/zenodo.12538729" httpUri="https://zenodo.org/record/12538729/files/figure.png" pageId="4" pageNumber="65">7</figureCitation>
|
||
) as a result of transverse wave formation. The vesicular nucleus was 2–7 µm in diameter and contained a centrally located nucleolus of 1–5 µm in diameter (arrowheads in
|
||
<figureCitation id="12BA7595FFA4FF88FBAEFB2FFAE86504" box="[1146,1296,1259,1285]" captionStart-0="Fig" captionStart-1="Fig" captionStart-2="Fig" captionStart-3="Fig" captionStartId-0="6.[132,167,1694,1716]" captionStartId-1="7.[125,160,1721,1743]" captionStartId-2="9.[127,162,1135,1157]" captionStartId-3="9.[125,160,1695,1717]" captionTargetBox-2="[127,1452,221,1108]" captionTargetId-0="figure-8@5.[180,1401,222,1933]" captionTargetId-1="figure-9@8.[142,1454,223,1930]" captionTargetId-2="figure-7@9.[127,1452,221,1108]" captionTargetId-3="figure-10@10.[177,1411,223,1939]" captionTargetPageId-0="5" captionTargetPageId-1="8" captionTargetPageId-2="9" captionTargetPageId-3="10" captionText-0="Fig. 1.A – 18S rDNA maximum likelihood phylogeny of Vannella, including Microcystis–associated strains (in bold). ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of V. planctonica (strainsA2FBB: 2, 4, 5, 9–13 andA4P4ZHB:1, 3, 6–8) showing locomotive trophozoites (1–8), a grazing amoeba with a Microcystis cell inside a food vacuole (9), floating forms (10–11) and a cyst stage (12–13). C – LM pictures of V. simplex (strain A17WVB) showing the floating form (14), locomotive amoebae (15–18) with the presence of food vacuoles containing partly digested Microcystis cells (17), posteriorly adhered fecal pellets (15–16), a long flagellum-like pseudopodium encircling a Microcystis cell (17–18) and grazing amoebae on a colony of Microcystis aeruginosa with expanded (arrow) or contracted (arrowhead) flagellum-like pseudopodia visible in some of the trophozoites (19). The presence of a contracticle vacuole, a nucleus or a cyst opening is indicated by a black arrow, a black arrowhead or a white arrow respectively. Scale bars: 20 µm." captionText-1="Fig. 2. A – 18S rDNA maximum likelihood phylogeny of Korotnevella, including Microcystis-associated strains (in bold). ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of K. jeppesenii (strain A1JEPDK) showing trophozoites in non-oriented movement (1–5), during locomotion (6, 8) and during grazing on Microcystis aeruginosa cells (7) with the arrow indicating a lacuna-containing nucleolus. C – LM pictures of K. pelagolacustris (strains A8WVB (7, 12), A16WVB (6, 8, 11), A21WVB (1, 13), A54WVB (3–5) and A1LMS (2, 9, 10)) showing trophozoites in different stages of non-oriented movement (1–4, 6), the locomotive form (8–12), grazing amoebae with Microcystis cells inside food vacuoles (5, 10), the cyst stage (7) and the floating form (13).Arrows indicate the nucleus with clearly visible nucleolus (in 8, 10, 12). Scale bars: 20 µm." captionText-2="Fig. 3. TEM pictures of the scales on the cell surface of Korotnevella jeppesenii (a, c) and K. pelagolacustris (b, d) trophozoites viewed from top (a, b) and side (c, d). Scale bars: 500 nm." captionText-3="Fig. 4. A – 18S rDNA maximum likelihood phylogeny of the family Hartmannellidae, including Microcystis-associated strains of Copromyxa (in bold). ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of C. microcystidis showing locomotive trophozoites (1–7, 10) containing a pronounced hyaline cap (white arrowheads), a vesicular nucleus with globular nucleolus (white arrows), big crystals (black arrows) and Microcystis cells inside food vacuoles (black arrowheads), non-oriented moving trophozoites (8–9, 11), a grazing amoeba capturing a Microcystis cell (12–14), the floating form (15) and a double-walled cyst stage (16). C – LM pictures of C. vandevyveri showing locomotive trophozoites (1, 6–9), trophozoites during non-oriented movement (2–5, 10–12) and floating forms (13–15). Black arrows indicate the presence of small, refractive crystals in the cytoplasm, the white arrow shows the vesicular nucleus with a globular lacuna-containing nucleolus, white arrowheads indicate a pronounced hyaline cap. Ingested Microcystis cells are visible in (9) and (11). Scale bars: 20 µm." figureDoi-0="http://doi.org/10.5281/zenodo.12538716" figureDoi-1="http://doi.org/10.5281/zenodo.12538718" figureDoi-2="http://doi.org/10.5281/zenodo.12538720" figureDoi-3="http://doi.org/10.5281/zenodo.12538723" httpUri-0="https://zenodo.org/record/12538716/files/figure.png" httpUri-1="https://zenodo.org/record/12538718/files/figure.png" httpUri-2="https://zenodo.org/record/12538720/files/figure.png" httpUri-3="https://zenodo.org/record/12538723/files/figure.png" pageId="4" pageNumber="65">Figs 1B 1–4</figureCitation>
|
||
). Usually one small contractile vacuole was present posteriorly (black arrows in
|
||
<figureCitation id="12BA7595FFA4FF88FC7AFAF4FBD1654B" box="[942,1065,1328,1354]" captionStart="Fig" captionStartId="6.[132,167,1694,1716]" captionTargetId="figure-8@5.[180,1401,222,1933]" captionTargetPageId="5" captionText="Fig. 1.A – 18S rDNA maximum likelihood phylogeny of Vannella, including Microcystis–associated strains (in bold). ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of V. planctonica (strainsA2FBB: 2, 4, 5, 9–13 andA4P4ZHB:1, 3, 6–8) showing locomotive trophozoites (1–8), a grazing amoeba with a Microcystis cell inside a food vacuole (9), floating forms (10–11) and a cyst stage (12–13). C – LM pictures of V. simplex (strain A17WVB) showing the floating form (14), locomotive amoebae (15–18) with the presence of food vacuoles containing partly digested Microcystis cells (17), posteriorly adhered fecal pellets (15–16), a long flagellum-like pseudopodium encircling a Microcystis cell (17–18) and grazing amoebae on a colony of Microcystis aeruginosa with expanded (arrow) or contracted (arrowhead) flagellum-like pseudopodia visible in some of the trophozoites (19). The presence of a contracticle vacuole, a nucleus or a cyst opening is indicated by a black arrow, a black arrowhead or a white arrow respectively. Scale bars: 20 µm." figureDoi="http://doi.org/10.5281/zenodo.12538716" httpUri="https://zenodo.org/record/12538716/files/figure.png" pageId="4" pageNumber="65">Figs 1B 1</figureCitation>
|
||
,
|
||
<figureCitation id="12BA7595FFA4FF88FBE8FAF4FBB5654B" box="[1084,1101,1328,1354]" captionStart="Fig" captionStartId="15.[124,159,1777,1799]" captionTargetId="figure-9@16.[190,1408,222,1933]" captionTargetPageId="16" captionText="Fig. 7. A – 18S rDNA maximum likelihood phylogeny of Cochliopodium, including the Microcystis-associated strain (in bold). ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of locomotive amoebae clearly showing the surrounding hyaline sheet (black arrows) punctuated with microscales (best visible in 8), a few small subpseudopodia (black arrowheads) and trailing adhesive uroidal filaments (white arrows) (1–5), a tropho- zoite during and just after grazing on Microcystis cells showing a prominent fringe of folded hyaloplasm (white arrowheads) (6–9), the bell-shaped form on colonies of Microcystis aeruginosa (10–11) and floating amoebae (12–13). Scale bars: 20 µm." figureDoi="http://doi.org/10.5281/zenodo.12538729" httpUri="https://zenodo.org/record/12538729/files/figure.png" pageId="4" pageNumber="65">7</figureCitation>
|
||
,
|
||
<figureCitation id="12BA7595FFA4FF88FB89FAF4FB95654B" box="[1117,1133,1328,1354]" captionStart="Fig" captionStartId="17.[125,160,1778,1800]" captionTargetId="figure-10@18.[148,1448,222,1933]" captionTargetPageId="18" captionText="Fig. 8. A – 18S rDNA maximum likelihood phylogeny of Angulamoeba. Sequences new for this study are indicated in bold. ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of Angulamoeba microcystivorans (strains A1WVB (1, 2, 4) and A4WVB (3)) showing the characteristic rusty-colored plasmodium-like mucilage matrix with embedded trophozoites devouring a culture of Microcystis aeruginosa colonies (1–2), multi-vacuolated trophozoites that are seemingly connected by their filopodia (3) and amoeba-flagellates in different stages of transition (4). Scale bars: 200 µm (1), 100 µm (2), 20 µm (3, 4)." figureDoi="http://doi.org/10.5281/zenodo.12538733" httpUri="https://zenodo.org/record/12538733/files/figure.png" pageId="4" pageNumber="65">8</figureCitation>
|
||
), with up to 3 vacuoles in some individuals. Floating amoebae were either radial shaped, 10–35 µm in diameter, symmetric, displaying radially 5–11 pointed and basally thickened (up to 6 µm in width) pseudopodia up to 50 µm in length, or sometimes irregularly shaped with only one to a few thick and round-tipped pseudopodia (
|
||
<figureCitation id="12BA7595FFA4FF88FB65FA3BFAEB6618" box="[1201,1299,1535,1561]" captionStart="Fig" captionStartId="6.[132,167,1694,1716]" captionTargetId="figure-8@5.[180,1401,222,1933]" captionTargetPageId="5" captionText="Fig. 1.A – 18S rDNA maximum likelihood phylogeny of Vannella, including Microcystis–associated strains (in bold). ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of V. planctonica (strainsA2FBB: 2, 4, 5, 9–13 andA4P4ZHB:1, 3, 6–8) showing locomotive trophozoites (1–8), a grazing amoeba with a Microcystis cell inside a food vacuole (9), floating forms (10–11) and a cyst stage (12–13). C – LM pictures of V. simplex (strain A17WVB) showing the floating form (14), locomotive amoebae (15–18) with the presence of food vacuoles containing partly digested Microcystis cells (17), posteriorly adhered fecal pellets (15–16), a long flagellum-like pseudopodium encircling a Microcystis cell (17–18) and grazing amoebae on a colony of Microcystis aeruginosa with expanded (arrow) or contracted (arrowhead) flagellum-like pseudopodia visible in some of the trophozoites (19). The presence of a contracticle vacuole, a nucleus or a cyst opening is indicated by a black arrow, a black arrowhead or a white arrow respectively. Scale bars: 20 µm." figureDoi="http://doi.org/10.5281/zenodo.12538716" httpUri="https://zenodo.org/record/12538716/files/figure.png" pageId="4" pageNumber="65">Figs 1B</figureCitation>
|
||
10–11) or in a few cases even without pseudopodia. Fecal pellets, varying in size between 5 and 13 µm, were present in the culture medium but did not tend to adhere posteriorly. Elliptical to ovoid cysts were observed a few times in old cultures. These had a thin wall (0.5 µm) and a diameter of 11–32 µm. Each had either a round opening of about 6 µm in diameter (white arrow in
|
||
<figureCitation id="12BA7595FFA4FF88FAFAF935FA7F670A" box="[1326,1415,1777,1803]" captionStart="Fig" captionStartId="6.[132,167,1694,1716]" captionTargetId="figure-8@5.[180,1401,222,1933]" captionTargetPageId="5" captionText="Fig. 1.A – 18S rDNA maximum likelihood phylogeny of Vannella, including Microcystis–associated strains (in bold). ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of V. planctonica (strainsA2FBB: 2, 4, 5, 9–13 andA4P4ZHB:1, 3, 6–8) showing locomotive trophozoites (1–8), a grazing amoeba with a Microcystis cell inside a food vacuole (9), floating forms (10–11) and a cyst stage (12–13). C – LM pictures of V. simplex (strain A17WVB) showing the floating form (14), locomotive amoebae (15–18) with the presence of food vacuoles containing partly digested Microcystis cells (17), posteriorly adhered fecal pellets (15–16), a long flagellum-like pseudopodium encircling a Microcystis cell (17–18) and grazing amoebae on a colony of Microcystis aeruginosa with expanded (arrow) or contracted (arrowhead) flagellum-like pseudopodia visible in some of the trophozoites (19). The presence of a contracticle vacuole, a nucleus or a cyst opening is indicated by a black arrow, a black arrowhead or a white arrow respectively. Scale bars: 20 µm." figureDoi="http://doi.org/10.5281/zenodo.12538716" httpUri="https://zenodo.org/record/12538716/files/figure.png" pageId="4" pageNumber="65">Fig. 1B</figureCitation>
|
||
12) or a bone-shaped narrow slit of about 11 µm in length (white arrow in
|
||
<figureCitation id="12BA7595FFA4FF88FC39F8F2FBBD6751" box="[1005,1093,1846,1872]" captionStart="Fig" captionStartId="6.[132,167,1694,1716]" captionTargetId="figure-8@5.[180,1401,222,1933]" captionTargetPageId="5" captionText="Fig. 1.A – 18S rDNA maximum likelihood phylogeny of Vannella, including Microcystis–associated strains (in bold). ML bootstrap values respectively posterior probabilities are shown at the nodes. GenBank accession numbers are given together with the species names. B – LM pictures of V. planctonica (strainsA2FBB: 2, 4, 5, 9–13 andA4P4ZHB:1, 3, 6–8) showing locomotive trophozoites (1–8), a grazing amoeba with a Microcystis cell inside a food vacuole (9), floating forms (10–11) and a cyst stage (12–13). C – LM pictures of V. simplex (strain A17WVB) showing the floating form (14), locomotive amoebae (15–18) with the presence of food vacuoles containing partly digested Microcystis cells (17), posteriorly adhered fecal pellets (15–16), a long flagellum-like pseudopodium encircling a Microcystis cell (17–18) and grazing amoebae on a colony of Microcystis aeruginosa with expanded (arrow) or contracted (arrowhead) flagellum-like pseudopodia visible in some of the trophozoites (19). The presence of a contracticle vacuole, a nucleus or a cyst opening is indicated by a black arrow, a black arrowhead or a white arrow respectively. Scale bars: 20 µm." figureDoi="http://doi.org/10.5281/zenodo.12538716" httpUri="https://zenodo.org/record/12538716/files/figure.png" pageId="4" pageNumber="65">Fig. 1B</figureCitation>
|
||
13).
|
||
</paragraph>
|
||
<paragraph id="8A3E6910FFA4FF8AFC82F89CFE1F63D5" blockId="4.[816,1463,223,1941]" lastBlockId="6.[132,779,223,980]" lastPageId="6" lastPageNumber="67" pageId="4" pageNumber="65">
|
||
Thirty
|
||
<taxonomicName id="4D811293FFA4FF88FC79F89CFBEC6773" ID-CoL="85VM" authority="Bovee, 1965" box="[941,1044,1880,1906]" class="Discosea" family="Vannellidae" genus="Vannella" kingdom="Protozoa" order="Vannellida" pageId="4" pageNumber="65" phylum="Amoebozoa" rank="genus">
|
||
<emphasis id="B8F5B502FFA4FF88FC79F89CFBEC6773" box="[941,1044,1880,1906]" italics="true" pageId="4" pageNumber="65">Vannella</emphasis>
|
||
</taxonomicName>
|
||
species are currently recognized, 24 from marine and brackish, 4 from freshwater and 2 from soil environments (
|
||
<bibRefCitation id="EE1014E1FFA6FF8AFE1EFF1BFD3E60F8" author="Smirnov A. V. & Nassonova E. S. & Chao E. & Cavalier-Smith T." box="[458,710,223,250]" pageId="6" pageNumber="67" pagination="295 - 324" refId="ref20038" refString="Smirnov A. V., Nassonova E. S., Chao E., Cavalier-Smith T. (2007) Phylogeny, evolution and taxonomy of vannellid amoebae. Protist 158: 295 - 324" type="journal article" year="2007">
|
||
Smirnov
|
||
<emphasis id="B8F5B502FFA6FF8AFDE8FF24FD8C60F8" box="[572,628,223,249]" italics="true" pageId="6" pageNumber="67">et al</emphasis>
|
||
. 2007
|
||
</bibRefCitation>
|
||
). Our phylogenetic analysis placed 4 of our vannellid strains in a clade containing several strains of
|
||
<taxonomicName id="4D811293FFA6FF8AFDB3FEE1FF4D6163" authorityName="Bovee" authorityYear="1965" class="Discosea" family="Vannellidae" genus="Vannella" kingdom="Protozoa" order="Vannellida" pageId="6" pageNumber="67" phylum="Amoebozoa" rank="species" species="simplex">
|
||
<emphasis id="B8F5B502FFA6FF8AFDB3FEE1FF4D6163" italics="true" pageId="6" pageNumber="67">Vannella simplex</emphasis>
|
||
</taxonomicName>
|
||
, a species with a wide geographically distribution, isolated from both freshwater and brackish habitats and genetically rather variable (
|
||
<bibRefCitation id="EE1014E1FFA6FF8AFE1AFE49FD3D61A6" author="Smirnov A. V. & Nassonova E. & Holzmann M. & Pawlowski J." box="[462,709,397,424]" pageId="6" pageNumber="67" pagination="367 - 377" refId="ref19916" refString="Smirnov A. V., Nassonova E., Holzmann M., Pawlowski J. (2002) Morphological, ecological and molecular studies of Vannella simplex Wohlfarth-Bottermann 1960 (Lobosea, Gymnamoebia), with a new diagnosis of this species. Protist 153: 367 - 377" type="journal article" year="2002">
|
||
Smirnov
|
||
<emphasis id="B8F5B502FFA6FF8AFDEBFE4AFD8E61A6" box="[575,630,397,423]" italics="true" pageId="6" pageNumber="67">et al</emphasis>
|
||
. 2002
|
||
</bibRefCitation>
|
||
,
|
||
<bibRefCitation id="EE1014E1FFA6FF8AFD07FE4AFE9761CB" author="Nassonova E. & Smirnov A. & Fahrni J. & Pawlowski J." pageId="6" pageNumber="67" pagination="102 - 115" refId="ref18618" refString="Nassonova E., Smirnov A., Fahrni J., Pawlowski J. (2010) Barcoding amoebae: comparison of SSU, ITS and COI genes as tools for molecular identification of naked lobose amoebae. Protist 161: 102 - 115" type="journal article" year="2010">
|
||
Nassonova
|
||
<emphasis id="B8F5B502FFA6FF8AFF30FE75FEE461CB" box="[228,284,432,458]" italics="true" pageId="6" pageNumber="67">et al</emphasis>
|
||
. 2010
|
||
</bibRefCitation>
|
||
). Since our strains also share the relatively large cell size and the coverage by fecal pellets, both typical for
|
||
<taxonomicName id="4D811293FFA6FF8AFEAEFE32FE09620E" authorityName="Bovee" authorityYear="1965" box="[378,497,501,527]" class="Discosea" family="Vannellidae" genus="Vannella" kingdom="Protozoa" order="Vannellida" pageId="6" pageNumber="67" phylum="Amoebozoa" rank="species" species="simplex">
|
||
<emphasis id="B8F5B502FFA6FF8AFEAEFE32FE09620E" box="[378,497,501,527]" italics="true" pageId="6" pageNumber="67">V. simplex</emphasis>
|
||
</taxonomicName>
|
||
isolates (
|
||
<bibRefCitation id="EE1014E1FFA6FF8AFDB5FE31FF3B6233" author="Smirnov A. V. & Nassonova E. & Holzmann M. & Pawlowski J." pageId="6" pageNumber="67" pagination="367 - 377" refId="ref19916" refString="Smirnov A. V., Nassonova E., Holzmann M., Pawlowski J. (2002) Morphological, ecological and molecular studies of Vannella simplex Wohlfarth-Bottermann 1960 (Lobosea, Gymnamoebia), with a new diagnosis of this species. Protist 153: 367 - 377" type="journal article" year="2002">
|
||
Smirnov
|
||
<emphasis id="B8F5B502FFA6FF8AFD1BFE32FCFB620E" box="[719,771,501,527]" italics="true" pageId="6" pageNumber="67">et al</emphasis>
|
||
. 2002
|
||
</bibRefCitation>
|
||
), we believe they indeed belong to this species. Our remaining vannellid strains were much smaller in size, lacked the coverage by fecal pellets and formed a separate clade with uncultured clones or unidentified
|
||
<taxonomicName id="4D811293FFA6FF8AFF50FD67FF1362BC" authority="Bovee, 1965" box="[132,235,675,701]" class="Discosea" family="Vannellidae" genus="Vannella" kingdom="Protozoa" order="Vannellida" pageId="6" pageNumber="67" phylum="Amoebozoa" rank="genus">
|
||
<emphasis id="B8F5B502FFA6FF8AFF50FD67FF1362BC" box="[132,235,675,701]" italics="true" pageId="6" pageNumber="67">Vannella</emphasis>
|
||
</taxonomicName>
|
||
’s as their closest relatives. They were clearly separated from two other freshwater species,
|
||
<taxonomicName id="4D811293FFA6FF8AFD14FD03FF346302" class="Discosea" family="Vannellidae" genus="Vannella" kingdom="Protozoa" order="Vannellida" pageId="6" pageNumber="67" phylum="Amoebozoa" rank="species" species="miroides">
|
||
<emphasis id="B8F5B502FFA6FF8AFD14FD03FF346302" italics="true" pageId="6" pageNumber="67">V. miroides</emphasis>
|
||
</taxonomicName>
|
||
and
|
||
<taxonomicName id="4D811293FFA6FF8AFED7FD2EFEB56302" box="[259,333,745,771]" class="Discosea" family="Vannellidae" genus="Vannella" kingdom="Protozoa" order="Vannellida" pageId="6" pageNumber="67" phylum="Amoebozoa" rank="species" species="lata">
|
||
<emphasis id="B8F5B502FFA6FF8AFED7FD2EFEB56302" box="[259,333,745,771]" italics="true" pageId="6" pageNumber="67">V. lata</emphasis>
|
||
</taxonomicName>
|
||
. No sequence information is available for
|
||
<taxonomicName id="4D811293FFA6FF8AFF7BFCC8FEDE6327" box="[175,294,780,806]" class="Discosea" family="Vannellidae" genus="Vannella" kingdom="Protozoa" order="Vannellida" pageId="6" pageNumber="67" phylum="Amoebozoa" rank="species" species="cirifera">
|
||
<emphasis id="B8F5B502FFA6FF8AFF7BFCC8FEDE6327" box="[175,294,780,806]" italics="true" pageId="6" pageNumber="67">V. cirifera</emphasis>
|
||
</taxonomicName>
|
||
, the only other freshwater
|
||
<taxonomicName id="4D811293FFA6FF8AFDBEFCCFFD296324" authority="Bovee, 1965" box="[618,721,779,805]" class="Discosea" family="Vannellidae" genus="Vannella" kingdom="Protozoa" order="Vannellida" pageId="6" pageNumber="67" phylum="Amoebozoa" rank="genus">
|
||
<emphasis id="B8F5B502FFA6FF8AFDBEFCCFFD296324" box="[618,721,779,805]" italics="true" pageId="6" pageNumber="67">Vannella</emphasis>
|
||
</taxonomicName>
|
||
species known, however this species was announced a
|
||
<emphasis id="B8F5B502FFA6FF8AFD37FCEBFEE0636A" italics="true" pageId="6" pageNumber="67">nomen dubium</emphasis>
|
||
by
|
||
<bibRefCitation id="EE1014E1FFA6FF8AFE9EFC95FDB0636A" author="Smirnov A. V. & Nassonova E. & Holzmann M. & Pawlowski J." box="[330,584,849,876]" pageId="6" pageNumber="67" pagination="367 - 377" refId="ref19916" refString="Smirnov A. V., Nassonova E., Holzmann M., Pawlowski J. (2002) Morphological, ecological and molecular studies of Vannella simplex Wohlfarth-Bottermann 1960 (Lobosea, Gymnamoebia), with a new diagnosis of this species. Protist 153: 367 - 377" type="journal article" year="2002">
|
||
Smirnov
|
||
<emphasis id="B8F5B502FFA6FF8AFE6DFC96FE17636A" box="[441,495,849,875]" italics="true" pageId="6" pageNumber="67">et al</emphasis>
|
||
. (2002
|
||
</bibRefCitation>
|
||
,
|
||
<bibRefCitation id="EE1014E1FFA6FF8AFD83FC95FD6F636A" author="Smirnov A. V. & Nassonova E. S. & Chao E. & Cavalier-Smith T." box="[599,663,849,875]" pageId="6" pageNumber="67" pagination="295 - 324" refId="ref20038" refString="Smirnov A. V., Nassonova E. S., Chao E., Cavalier-Smith T. (2007) Phylogeny, evolution and taxonomy of vannellid amoebae. Protist 158: 295 - 324" type="journal article" year="2007">2007</bibRefCitation>
|
||
). For this reason and because of its specific ecology we declared the strains of our second lineage as belonging to a new species,
|
||
<taxonomicName id="4D811293FFA6FF8AFF3CFC7DFE1863D2" authorityName="Van Wichelen & Vanormelingen" authorityYear="2016" box="[232,480,953,979]" class="Discosea" family="Vannellidae" genus="Vannella" kingdom="Protozoa" order="Vannellida" pageId="6" pageNumber="67" phylum="Amoebozoa" rank="species" species="planctonica">
|
||
<emphasis id="B8F5B502FFA6FF8AFF3CFC7DFE1863D2" box="[232,480,953,979]" italics="true" pageId="6" pageNumber="67">Vannella planctonica</emphasis>
|
||
</taxonomicName>
|
||
.
|
||
</paragraph>
|
||
</subSubSection>
|
||
</treatment>
|
||
</document> |