212 lines
20 KiB
XML
212 lines
20 KiB
XML
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<mods:title id="ADB1954D10F3DF6F2790072462FFA37B">Italian wolves (Canis lupus italicus Altobello, 1921) and molecular detection of taeniids in the Foreste Casentinesi National Park, Northern Italian Apennines</mods:title>
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<mods:namePart id="6107DA384DEE7D71A1764BF74ECD3F78">Poglayen, Giovanni</mods:namePart>
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<mods:namePart id="B059505D8AA6407E1011BD1F2CA92E1E">Gori, Francesca</mods:namePart>
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<mods:namePart id="01EEBF2D0771D8071E66A9C92AD9E8CA">Caniglia, Romolo</mods:namePart>
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<mods:affiliation id="0CC2E2472D5ABF686EE88B23E547B570">Genetics Laboratory, Institute for Environmental Protection and Research (ISPRA), Ozzano dell’Emilia, Bologna, Italy</mods:affiliation>
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<mods:namePart id="9821A89BFD309CBDA91D011840A9609B">Galaverni, Marco</mods:namePart>
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<mods:namePart id="3781CEAC243F4BF0B42394CCD2F56C7B">Randi, Ettore</mods:namePart>
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<mods:affiliation id="5E5D09099B2054B1455D87FF9FF93046">Alma Mater Studiorum, Department of Veterinary Medical Science, Via Tolara di Sopra, 50 - 40064, Ozzano dell’Emilia, Bologna, Italy</mods:affiliation>
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<mods:affiliation id="18B714E8AB50D38BC24D3E4A159924C1">Institute of Parasitology, University of Zurich, Winterthurerstrasse 266 a, CH- 8057 Zurich, Switzerland</mods:affiliation>
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<mods:title id="DED1D5B42AB06378EA14FA287300458C">International Journal for Parasitology: Parasites and Wildlife</mods:title>
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<treatment id="03A1F61AFFA39E48275B7117FBEE78CE" ID-DOI="http://doi.org/10.5281/zenodo.10892131" ID-Zenodo-Dep="10892131" LSID="urn:lsid:plazi:treatment:03A1F61AFFA39E48275B7117FBEE78CE" httpUri="http://treatment.plazi.org/id/03A1F61AFFA39E48275B7117FBEE78CE" lastPageNumber="3" pageId="2" pageNumber="3">
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<subSubSection id="C3121487FFA39E48275B7117FE0F7C4D" box="[113,388,1796,1816]" pageId="2" pageNumber="3" type="nomenclature">
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<heading id="D0FFF060FFA39E48275B7117FE0F7C4D" box="[113,388,1796,1816]" fontSize="36" level="2" pageId="2" pageNumber="3" reason="3">
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<emphasis id="B97C9B1EFFA39E48275B7117FE0F7C4D" box="[113,388,1796,1816]" italics="true" pageId="2" pageNumber="3">
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2.5.
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<taxonomicName id="4C083C8FFFA39E48278B7117FF767C4D" ID-CoL="7NL8H" authorityName="Ludwig" authorityYear="1886" box="[161,253,1796,1816]" class="Cestoda" family="Taeniidae" kingdom="Animalia" order="Cyclophyllidea" pageId="2" pageNumber="3" phylum="Platyhelminthes" rank="family">Taeniidae</taxonomicName>
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identi
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<emphasis id="B97C9B1EFFA39E4826167116FEC37C4D" bold="true" box="[316,328,1797,1816]" italics="true" pageId="2" pageNumber="3">fi</emphasis>
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cation
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<paragraph id="8BB7470CFFA39E4827BB712EFE837CF1" blockId="2.[113,783,1853,1984]" pageId="2" pageNumber="3">
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<materialsCitation id="3B604D51FFA39E4827BB712EFE8F7CF1" collectionCode="ISPRA" pageId="2" pageNumber="3" specimenCount="130">
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From the
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<collectionCode id="ED19DFC9FFA39E4827D3712EFEBF7C05" box="[249,308,1853,1872]" pageId="2" pageNumber="3">ISPRA</collectionCode>
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genetic bank
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<specimenCount id="9D0E8C85FFA39E4826E0712EFDE97C05" box="[458,610,1853,1872]" count="130" pageId="2" pageNumber="3" type="generic">130 specimens</specimenCount>
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belonging to 54 wolves, chosen according to the genetic profile (not dog or wolf x dog hybrid) and to abundance of material, were examined for taeniidae eggs
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</materialsCitation>
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.
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</paragraph>
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<paragraph id="8BB7470CFFA39E4827BB71BEFBEE78CE" blockId="2.[113,783,1853,1984]" lastBlockId="2.[831,1501,179,923]" pageId="2" pageNumber="3">
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Up to
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<quantity id="4CF0EAE9FFA39E4827F971BEFF7E7C95" box="[211,245,1965,1984]" metricMagnitude="-3" metricUnit="kg" metricValue="2.0" pageId="2" pageNumber="3" unit="g" value="2.0">2 g</quantity>
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of feces were sieved in a filter (mesh 150
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<emphasis id="B97C9B1EFFA39E48259271BEFD4F7C95" bold="true" box="[696,708,1965,1984]" pageId="2" pageNumber="3">M</emphasis>
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m) and washed several times in a cup. The filtrate was centrifuged (1600 × g) for ten minutes and the pellet collected. Taeniid eggs were isolated from the pellet using the flotation and sieving method described by
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<bibRefCitation id="EF993AFDFFA39E48231F7714FA9F7A4F" author="Mathis, A. & Deplazes, P. & Eckert, J." box="[1077,1300,263,282]" pageId="2" pageNumber="3" pagination="219 - 222" refId="ref7932" refString="Mathis, A., Deplazes, P., Eckert, J., 1996. An improved test system for PCR-based specific detection of Echinococcus multilocularis eggs. J. Helmintol. 70, 219 - 222." type="journal article" year="1996">Mathis et al. (1996)</bibRefCitation>
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and subjected to morphological identification under an inverted microscope. In egg positive samples and in 14 of the negative samples, DNA extraction was carried out with the complete sieving fraction as described by ̆Stefani ć et al. (2004). The 14 negative samples were included as negative controls. A total of 69 samples were analyzed using a multiplex-PCR to discriminate between
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<taxonomicName id="4C083C8FFFA39E48223B77BEFA2A7A94" baseAuthorityName="Batsch" baseAuthorityYear="1786" box="[1297,1441,429,449]" class="Insecta" family="Pseudococcidae" genus="Echinococcus" kingdom="Animalia" order="Hemiptera" pageId="2" pageNumber="3" phylum="Arthropoda" rank="species" species="granulosus">
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<emphasis id="B97C9B1EFFA39E48223B77BEFA2A7A94" box="[1297,1441,429,449]" italics="true" pageId="2" pageNumber="3">E. granulosus</emphasis>
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</taxonomicName>
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and
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<taxonomicName id="4C083C8FFFA39E48241577DAFC507A88" baseAuthorityName="Leuckart" baseAuthorityYear="1863" box="[831,987,457,477]" class="Insecta" family="Pseudococcidae" genus="Echinococcus" kingdom="Animalia" order="Hemiptera" pageId="2" pageNumber="3" phylum="Arthropoda" rank="species" species="multilocularis">
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<emphasis id="B97C9B1EFFA39E48241577DAFC507A88" box="[831,987,457,477]" italics="true" pageId="2" pageNumber="3">E. multilocularis</emphasis>
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</taxonomicName>
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and other cestodes including
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<taxonomicName id="4C083C8FFFA39E48222677DAFAF07A88" box="[1292,1403,457,477]" class="Cestoda" family="Taeniidae" genus="Taenia" kingdom="Animalia" order="Cyclophyllidea" pageId="2" pageNumber="3" phylum="Platyhelminthes" rank="species" species="undetermined">
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||
<emphasis id="B97C9B1EFFA39E48222677DAFAC07A88" box="[1292,1355,457,477]" italics="true" pageId="2" pageNumber="3">Taenia</emphasis>
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spp.
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</taxonomicName>
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(
|
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<bibRefCitation id="EF993AFDFFA39E4822A377D9FC267AAC" author="Trachsel, D. & Deplazes, P. & Mathis, A." pageId="2" pageNumber="3" pagination="911 - 920" refId="ref8995" refString="Trachsel, D., Deplazes, P., Mathis, A., 2007. Identification of taeniid eggs in the feces from carnivores based on multiplex-PCR using targets in mitochondrial DNA. Parasitol 134, 911 - 920." type="journal article" year="2007">Trachsel et al., 2007</bibRefCitation>
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). To obtain clear sequences of the
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<taxonomicName id="4C083C8FFFA39E48222277F6FA0C7AAC" baseAuthorityName="Batsch" baseAuthorityYear="1786" box="[1288,1415,485,505]" class="Insecta" family="Pseudococcidae" genus="Echinococcus" kingdom="Animalia" order="Hemiptera" pageId="2" pageNumber="3" phylum="Arthropoda" rank="species" species="granulosus">
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<emphasis id="B97C9B1EFFA39E48222277F6FA9F7AAC" box="[1288,1300,485,505]" italics="true" pageId="2" pageNumber="3">E</emphasis>
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.
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<emphasis id="B97C9B1EFFA39E48223477F6FA0C7AAC" box="[1310,1415,485,505]" italics="true" pageId="2" pageNumber="3">granulosus</emphasis>
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</taxonomicName>
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positive samples, the PCR was repeated but only using
|
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<taxonomicName id="4C083C8FFFA39E4822207412FA037940" baseAuthorityName="Batsch" baseAuthorityYear="1786" box="[1290,1416,513,533]" class="Insecta" family="Pseudococcidae" genus="Echinococcus" kingdom="Animalia" order="Hemiptera" pageId="2" pageNumber="3" phylum="Arthropoda" rank="species" species="granulosus">
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<emphasis id="B97C9B1EFFA39E4822207412FA037940" box="[1290,1416,513,533]" italics="true" pageId="2" pageNumber="3">E. granulosus</emphasis>
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</taxonomicName>
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primers (Cest5 and Cest4) keeping the same conditions as described above, and therefore amplicons were sequenced. Another PCR targeting
|
||
<emphasis id="B97C9B1EFFA39E4824157447FCE4793D" box="[831,879,596,616]" italics="true" pageId="2" pageNumber="3">nad1</emphasis>
|
||
gene (
|
||
<bibRefCitation id="EF993AFDFFA39E4824967446FB64793C" author="Armua-Fernandez, M. T. & Nonaka, N. & Sakurai, T. & Nakamura, S. & Gottstein, B. & Deplazes, P. & Phiri, I. G. K. & Katakura, K. & Oku, Y." box="[956,1263,597,617]" pageId="2" pageNumber="3" pagination="84 - 89" refId="ref5626" refString="Armua-Fernandez, M. T., Nonaka, N., Sakurai, T., Nakamura, S., Gottstein, B., Deplazes, P., Phiri, I. G. K., Katakura, K., Oku, Y., 2011. Development of PCR / dot blot assay for specific detection and differentiation of taeniid cestode eggs in canids. Parasitol. Int. 60, 84 - 89." type="journal article" year="2011">Armua-Fernandez et al., 2011</bibRefCitation>
|
||
) was used in samples without clear sequencing results to confirm the species of those samples. The amplicons were directly sequenced after purification of the PCR products using the MinElute
|
||
<superScript id="7C7DEA44FFA39E4823D974B0FB7479E4" attach="left" box="[1267,1279,675,689]" fontSize="6" pageId="2" pageNumber="3">®</superScript>
|
||
PCR purification kit (Qiagen). Sequencing was performed by Synergene Biotech GmbH, Biotech Center
|
||
<collectingRegion id="49CC89EEFFA39E4824D074F2FBB779A1" box="[1018,1084,737,756]" country="Switzerland" name="Zuerich" pageId="2" pageNumber="3">Zurich</collectingRegion>
|
||
,
|
||
<collectingCountry id="F31F079CFFA39E48237D74F2FB4479A1" box="[1111,1231,737,756]" name="Switzerland" pageId="2" pageNumber="3">Switzerland</collectingCountry>
|
||
(http://www.synergenebiotech.com). Primer Cest5 and Cest5seq was used for non-
|
||
<taxonomicName id="4C083C8FFFA39E48228174EFFC1D7879" class="Insecta" family="Pseudococcidae" genus="Echinococcus" kingdom="Animalia" order="Hemiptera" pageId="2" pageNumber="3" phylum="Arthropoda" rank="genus">
|
||
<emphasis id="B97C9B1EFFA39E48228174EFFC1D7879" italics="true" pageId="2" pageNumber="3">Echinococcus</emphasis>
|
||
</taxonomicName>
|
||
cestodes including
|
||
<taxonomicName id="4C083C8FFFA39E482371750BFB427879" box="[1115,1225,792,812]" class="Cestoda" family="Taeniidae" genus="Taenia" kingdom="Animalia" order="Cyclophyllidea" pageId="2" pageNumber="3" phylum="Platyhelminthes" rank="species" species="undetermined">
|
||
<emphasis id="B97C9B1EFFA39E482371750BFB117879" box="[1115,1178,792,812]" italics="true" pageId="2" pageNumber="3">Taenia</emphasis>
|
||
spp.
|
||
</taxonomicName>
|
||
and Cest5 for
|
||
<taxonomicName id="4C083C8FFFA39E482272750BFA5C7879" baseAuthorityName="Batsch" baseAuthorityYear="1786" box="[1368,1495,792,812]" class="Insecta" family="Pseudococcidae" genus="Echinococcus" kingdom="Animalia" order="Hemiptera" pageId="2" pageNumber="3" phylum="Arthropoda" rank="species" species="granulosus">
|
||
<emphasis id="B97C9B1EFFA39E482272750BFAEF7879" box="[1368,1380,792,812]" italics="true" pageId="2" pageNumber="3">E</emphasis>
|
||
.
|
||
<emphasis id="B97C9B1EFFA39E482244750BFA5C7879" box="[1390,1495,792,812]" italics="true" pageId="2" pageNumber="3">granulosus</emphasis>
|
||
</taxonomicName>
|
||
, while primer
|
||
<emphasis id="B97C9B1EFFA39E4824E27527FC73781D" box="[968,1016,820,840]" italics="true" pageId="2" pageNumber="3">nad1</emphasis>
|
||
T-Rv for
|
||
<taxonomicName id="4C083C8FFFA39E4823647527FB37781D" box="[1102,1212,820,840]" class="Cestoda" family="Taeniidae" genus="Taenia" kingdom="Animalia" order="Cyclophyllidea" pageId="2" pageNumber="3" phylum="Platyhelminthes" rank="species" species="undetermined">
|
||
<emphasis id="B97C9B1EFFA39E4823647527FB06781D" box="[1102,1165,820,840]" italics="true" pageId="2" pageNumber="3">Taenia</emphasis>
|
||
spp.
|
||
</taxonomicName>
|
||
for amplicons obtained with multiplex-PCR and
|
||
<emphasis id="B97C9B1EFFA39E482331755CFBC77836" box="[1051,1100,847,867]" italics="true" pageId="2" pageNumber="3">nad1</emphasis>
|
||
PCR, respectively. Sequences were compared with the one present in GenBank using Blast tool (http:// www.blast.ncbi.nlm.nih.gov).
|
||
</paragraph>
|
||
</subSubSection>
|
||
</treatment>
|
||
</document> |