Abstract
Microsporidiosis (nosema disease) of the honeybee, Apis mellifera, has spread worldwide and caused heavy economic losses in apiculture. We obtained a spore isolate from worker ventriculi of A. mellifera colonies kept on the campus of National Taiwan University and sequenced the ribosomal genes. The entire length of the ribosomal DNA is about 3828 bp and the organization is similar to that of Nosema apis. However, the SSUrRNA, ITS, and LSUrRNA sequences have comparatively low identities with those of N. apis (92, 52, and 89%, respectively) and the SSUrRNA has a 99% identity with Nosema ceranae. These results indicate that this isolate is not N. apis, but N. ceranae. Moreover, the morphological characteristics are identical to those of N. ceranae. These results show that nosema disease of the honeybee, A. mellifera, may not be caused solely by the infection of N. apis.
Zusammenfassung
Nosema-Erkrankungen (Mikrosporidien) bei Honigbienen, Apis mellifera, sind weltweit verbreitet und verursachen erhebliche wirtschaftliche Schäden in der Imkerei. Die Nosemose verbreitete sich nach 1972 in ganz Taiwan und konnte in einer bis 1980 dauernden Untersuchung in sämtlichen Frühjahrs- und Herbstuntersuchungen nachgewiesen werden. Aufbauend auf diesen Untersuchungen amplifizierten und sequenzierten wir die ribosomalen Gene von Sporen aus dem Verdauungstrakt von Arbeiterinnen, die aus Bienenvölkern auf dem Campus der National Taiwan Universität stammten. Die vollständige Sequenz der ribosomalen DNA beträgt 3.828 Basenpaare und ähnelt der von Nosema apis (Abb. 1). Allerdings weist die SSUrRNA-Sequenz eine 99 %ige Übereinstimmung mit der von N. ceranae auf und die SSUrRNA-, ITS- und LSUrRNA-Sequenzen haben eine verhältnismäßig geringe Ähnlichkeit mit denen von N. apis (92, 52 bzw. 89 %). Zudem stimmen bei den untersuchten Sporenisolaten auch die morphologischen Charakteristika, die Größe der lebenden Sporen sowie die Struktur des Polfadens im Längsschnitt (Abb. 3) mit denen von N. ceranae überein. All diese Ergebnisse zeigen, dass diese Sporen nicht von N. apis sondern von N. ceranae stammen. Bei der phylogenetischen Analyse zeigen sowohl der SSUrRNAals auch der LSUrRNA-Stammbaum (Abb. 4), dass N. ceranae und N. apis phylogenetisch gut zu trennen sind. Daher muss für Nosema-Erkrankungen bei Honigbienen (A. mellifera) nicht ausschließlich N. apis verantwortlich sein. Der Erreger der Nosemose sollte in zukünftigen Untersuchungen daher eindeutig bestimmt werden.
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References
An J.K., Ho K.K. (1980) The seasonal variation of Nosema apis Zander in Taiwan, Honeybee Sci. 1, 157–158.
Baker M.D., Vossbrinck C.R., Maddox J.V., Undeen A.H. (1994) Phylogenetic relationships among Vairimorpha and Nosema species (Microspora) based on ribosomal RNA sequence data, J. Invertebr. Pathol. 64, 100–106.
Burges H.D., Canning E.U., Hulls I.K. (1974) Ultrastructure of Nosema oryzaephili and the taxanomic value of the polar filament, J. Invertebr. Pathol. 23, 135–139.
Canning E.U., Curry A., Cheney S., Lafranchi-Tristem N.J., Haque M.A. (1999) Vairimorpha imperfecta n. sp., a microsporidian exhibiting an abortive octosporous sporogony in Plutella xylostella L. (Lepidoptera: Yponomeutidae), Parasitology 119, 273–286.
Fries I. (1989) Observation on the development and transmission of Nosema apis Z. in the ventriculus of the honeybee, J. Apic. Res. 28, 107–117.
Fries I. (1997) Protozoa, in: Morse R.A. (Ed.), Honey bee pests, predators, and diseases, 3rd ed., A.I Root Company, Medina, Ohio, USA, pp. 57–76.
Fries I., Ekbom G. (1984) Nosema apis, sampling techniques and honey yield, J. Apic. Res. 23, 102–105.
Fries I., Feng F., da Silva A., Slemenda S.B., Pieniazek N.J. (1996) Nosema ceranae n. sp. (Microspora, Nosematidae), Morphological and Molecular Characterization of a Microsporidian Parasite of the Asian Honey bee Apis cerana (Hymenoptera, Apidae), Eur. J. Protistol. 32, 356–365.
Gatehouse H.S., Malone L.A. (1998) The ribosomal RNA gene region of Nosema apis (Microspora): DNA sequence for small and large subunit rRNA genes and evidence of a large tandem repeat unit size, J. Invertebr. Pathol. 71, 97–105.
Higes M., Martín R., Meana A. (2006) Nosema ceranae, a new microsporidian parasite in honeybees in Europe, J. Invertebr. Pathol. 92, 93–95.
Huang H.W., Lo C.F., Tseng C.C., Peng S.E., Chou C.M., Kou C.H. (1998) The small subunit ribosomal RNA gene sequence of Pleistophora anguillarum and the use of PCR primers of diagnostic detection of the parasite, J. Eukaryot. Microbiol. 45, 556–560.
Huang W.F., Tsai S.J., Lo C.F., Soichi Y., Wang C.H. (2004) The novel organization and complete sequence of the ribosomal gene of Nosema bombycis, Fung. Genet. Biol. 41, 473–481.
Keeling P.J., Macfadden G.I. (1998) Origins of microsporidia, Trends Microbiol. 6, 19–23.
Larsson J.I.R. (2005) Fixation of microsporidian spores for electron microscopy, J. Invertebr. Pathol. 90, 47–50.
Lui T.P. (1973) The fine structure of frozen-etched spore of Nosema apis Zander, Tissue and Cell 5, 315–322.
Lui T.P., Lui H.J. (1974) Evaluation of some morphological characteristics of spore from two species of microsporidia by scanning electron microscope and frozen-etching techniques, J. Morphol. 143, 337–339.
Matheson A. (1996) World bee health update 1996, Bee World 77, 45–51.
Müller A., Trammer T., Ghioralia G., Seitz H.M., Diehl V., Franzen E. (2000) Ribosomal RNA of Nosema algerae and phylogenetic relationship to other microsporidia, Parasitol. Res. 86, 18–23.
Peyretaillade E., Biderre C., Peyret P., Duffieux F., Metenier G., Gouy M., Michot B., Vivares C.P. (1998) Microsporidian Encephalitozoon cuniculi, a unicellular eukaryote with an unusual chromosomal dispersion of ribosomal genes and a LSUrRNA reduced to the universal core, Nucleic Acids Res. 26, 3513–3520.
Reynolds E.S. (1963) The use of lead citrate at high pH as an electron-opaque stain in electron microscopy, J. Cell Biol. 17, 208–212.
Shyamala H., Ames G.F. (1989) Genome walking by single-specific-primer polymerase chain reaction: SSP-PCR, Gene 84, 1–8.
Singh Y. (1975) Nosema in Indian honey bee (Apis cerana indica), Am. Bee J. 115, 59.
Slamovits C.H., Willams B.A.P., Keeling P.J. (2004) Transfer of Nosema locustae (Microsporidia) to Antonospora locustae n. comb. Based on molecular and ultrastructure data, J. Eukaryot. Microbiol. 51, 207–213.
Swofford D.L. (2003) PAUP*, Phylogenetic analysis using parasimony (* and other methods), Sinauer Associates, Sunderland, MA.
Thompson J.D., Gibson T.J., Plewniak F., Jeanmougin F., Higgins D.G. (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools, Nucleic Acids Res. 25, 4876–4882.
Tsai S.J., Kou G.H., Lo C.F., Wang C.H. (2002) Complete sequence and structure of ribosomal RNA gene of Heterosporis anguillarum, Dis. Aquat. Org. 49, 199–206.
Tsai S.J., Lo C.F., Soichi Y., Wang C.H. (2003) The characterization of Microsporidian isolates (Nosematidae: Nosema) from five important lepidopteran pests in Taiwan, J. Invertebr. Pathol. 83, 51–59.
Tsai S.J., Huang W.F., Wang C.H. (2005) Complete sequence and gene organization of Nosema spodopterae rRNA gene, J. Eukaryot. Microbiol. 52, 52–54.
Undeen A.H., Cockburn A.F. (1989) The extraction of DNA from microsporidia spores, J. Invertebr. Pathol. 54, 132–133.
Van de Peer Y., De Rijk P., Wuyts J., Winkelmans T., De Wachter R. (2000) The Europen small subunit ribosomal RNA database, Nucleic Acids Res. 28, 175–176.
Vossbrinck C.R., Baker M.D., Didier E.S., Debrunner-Vossbrinck B.A., Shadduck J.A. (1993) Ribosomal DNA sequences of Encephalitozoon hellem and Encephalitozoon cuniculi: species identification and phylogentic construction, J. Eukaryot. Microbiol. 40, 354–362.
Weiss L.M., Vossbrinck C.R. (1999) Molecular biology, molecular phylogeny, and molecular diagnostic approaches to the microsporidia, in: Wittner M., Weiss L.M. (Eds.), The Microsporidia and Microsporidiosis, American Society for Microbiology, Washington, DC, pp. 129–171.
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Huang, WF., Jiang, JH., Chen, YW. et al. A Nosema ceranae isolate from the honeybee Apis mellifera . Apidologie 38, 30–37 (2007). https://doi.org/10.1051/apido:2006054
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DOI: https://doi.org/10.1051/apido:2006054