Balantioides coli is a ciliated protozoon that inhabits the intestine of pigs, non-human primates and humans. Light microscopy studies have described over 50 species of the genus Balantioides but their validity is in doubt. Due to the limited information about this genus, this study is aimed to identify morphological characteristics of Balantioides coli isolated using fluorescence microscopy and both scanning (SEM) and transmission electron microscopy (TEM). Trophozoites isolated from the feces of pig and macaque were washed and subjected to centrifugation. These cells were fixed with paraformaldehyde for immunofluorescence. Other aliquots of these trophozoites were fixed with glutaraldehyde, post fixed with osmium tetroxide and processed for SEM and TEM. Immunofluorescence studies revealed microtubules with a longitudinal distribution to the main axis of the parasite and in the constitution of cilia. SEM demonstrated a high concentration of cilia covering the oral apparatus and a poor presence of such structures in cytopyge. TEM revealed in the plasma membrane, several associated structures were observed to delineate the cellular cortex and mucocysts. The cytoskeleton of the oral region was observed in detail and had an organization pattern consisting of microtubules, which formed files and nematodesmal networks. Organelles such as hydrogenosomes like and peroxisomes were observed close to the cortex. Macronuclei were observed, but structures that were consistent with micronuclei were not identified. Ultrastructural morphological analysis of isolates confirms its similarity to Balantioides coli. In this study were identified structures that had not yet been described, such as hydrogenosomes like and cytoskeletal structures.
Author’s Contributions: Collection of the material and maintenance in culture: ASB, LVD, CMAU, OMPB, MRRA. Processing the material before description: ASB, LVD, HSB, SMOS. Morphological description: ASB, HSB. Draft of the manuscript written by: ASB, HSB. Revision of the manuscript: HSB, LVD, CMAU, OMPB, MRRA. ASB participated in all stages of production of this manuscript, because this is part of his PhD thesis. All authors read and approved the final version of the manuscript.
Ethical standards: All applicable federal, state, and institutional guidelines for the care and use of animals were followed in this study.
Conflict of interest: The authors declare that they have no conflict of interest.
The authors would like to acknowledge the contribution made by the Research Support Foundation of the State of Rio de Janeiro (FAPERJ), through grant number E26/110 440/2012. The authors thank too, Dr. Saulo Cabral Bourguignon of the Federal Fluminense University Biology Institute for supporting material and the Rudolph Barth Electron Microscopy Platform of Oswaldo Cruz Institute to produce images.
Barbosa A.S., Bastos O.M.P., Uchôa C.M.A., Pissinatti A., Filho P.R.F., Dib L.V., et al. 2015. Isolation and maintenance of Balantidium coli (Malmsteim, 1857) cultured from fecal samples of pigs and non-human primates. Veterinary Parasitology, 210, 240–245. 10.1016/j.vetpar.2015.03.030Search in Google Scholar PubMed
Barbosa A.S., Ponce-Gordo F., Dib L.V., Uchôa C.M.A., Bastos O.M.P., Pissinatti A., Amendoeira M.R.R. 2017. First molecular characterization of Balantioides coli (Malmsten, 1857) isolates maintained in vitro culture and from feces of captive animals, Rio de Janeiro, Brazil. Veterinary Parasitology: Regional Studies and Reports, 10, 102–113. https://doi.org/10.1016/j.vprsr.2017.08.01410.1016/j.vprsr.2017.08.014Search in Google Scholar PubMed
Chistyakova L.V., Kostygov A.Y., Kornillova A.O., Yurchenko V. 2014. Reisolation and redescription of Balantidium duodeni Stein, 1867 (Litostomatea, Trichostomatia). Parasitology Research, 113, 4207–15. 10.1007/s00436-014-4096-1Search in Google Scholar PubMed
Frasson A.P., Vieira P.B., De Carli G.A., Tasca T. 2010. Giardia lamblia: Distribuição de microtúbulos no citoesqueleto de trofozoítos e cistos utilizando taxóide fluorescente. Revista de Patologia Tropical, 39, 21–32. https://doi.org/10.5216/rpt.v39i1.949510.5216/rpt.v39i1.9495Search in Google Scholar
Grim J.N., Clements K.D., Byfield T. 2002. New species of Balantidium and Paracichlidotherus (Ciliophora) inhabiting the intestines of four surgeonfish species form the Tuvalu islands, Pacific ocean. The Journal of Eukaryotic Microbiology, 49, 146–153. 10.1111/j.1550-7408.2002.tb00359.xSearch in Google Scholar PubMed
Grim J.N., Buonanno F. 2009. A re-description of the ciliate genus and type species Balantidium entozoon. European Journal of Protistology, 45, 174–182. 10.1016/j.ejop.2008.10.001Search in Google Scholar PubMed
Hegner R. 1934. Specifity in the genus Balantidium based on size and shape of body and macronucleus, with descriptions of six new species. American Journal of Hygiene, 19, 38–6710.1093/oxfordjournals.aje.a117992Search in Google Scholar
Huttenlauch I., Peck R.K., Stick R. 1998. Articulins and epiplasmins: two distinct classes of cytoskeletal proteins of the membrane skeleton in protists. Journal of Cell Science, 111, 3367 – 337810.1242/jcs.111.22.3367Search in Google Scholar PubMed
Jones W.R. 1946. The experimental infection of rats with Entamoeba histolytica; Method for Evaluating with the anti-amoebic properties of new compounds. Annals of Tropical Medicine and Parasitology, 40, 130–140. http://dx.doi.org/10.1080/00034983.1946.1168527010.1080/00034983.1946.11685270Search in Google Scholar PubMed
King R.L., Beams H.W., Tahmisian T.N., Devine R.L. 1961. The ciliature and infraciliature of Nyctotherus ovalis Leidy. The Journal of Protozoology, 8, 98–111. 10.1111/j.1550-7408.1961.tb01189.xSearch in Google Scholar
Krascheninnikow S., Wenrich D.H. 1958. Some observations on the morphology and Division of Balantidium coli and Balantidium caviae. The Journal of Protozoology, 5, 196–202. 10.1111/j.1550-7408.1958.tb02551.xSearch in Google Scholar
Li M., Wang J., Zhang J., Gu Z., Ling F., Ke X., Gong X. 2008. First report of two Balantidium species from the Chinese giant salamander, Andrias davidianus: Balantidium sinensis Nie 1935 and Balantidium andianusis sp. Parasitology Research, 102, 605–611. 10.1007/s00436-007-0795-1Search in Google Scholar PubMed
Lynn D.H. (Ed.) 2008. The Ciliated protozoa. Characterization Classification and Guide to the Literature. Third Ed. Springer, New York, pp. 638Search in Google Scholar
Ming L., Chong W., Grim J.N., Jianguo W., Xiaoning G. 2014. Ultrastructural study of Balantidium ctenopharyngodoni Chen, 1955 (Class: Litostomatea) form China with an emphasis on its vestibulum. Acta Parasitologica, 59, 413–419. 10.2478/s11686-014-0262-ySearch in Google Scholar PubMed
Nilles-Bije M.L., Rivera W.L. 2010. Ultrastructural and molecular characterization of Balantidium coli isolated in the Philippines. Parasitology Research, 106, 387–394. 10.1007/s00436-009-1673-9Search in Google Scholar PubMed
Novikoff A.B. 1969. Visualization of peroxisomes (microbodies) and mitochondria with diaminobenzidine. The Journal Histochemistry and Cytochemistry, 17, 675–680. 10.1177/17.10.675Search in Google Scholar PubMed
Pavlova E.A. 1938. Sur les Méthodes de la culture d’Entamoeba histolytica. Meditsinskaia Parazitologiia I Parazitarnye Bolezni, 7, 224–227. (In French)Search in Google Scholar
Pomajbiková K., Oboromik M., Horák A., Petrzelkova K.J., Grim J.N., Levecke B., Todd A., Mulama M., Kiyang J., Modrý D. 2013. Novel Insights into the genetic diversity of Balantidium and Balantidium-like cyst – forming ciliates. Plos Neglected Tropical Disease, 7, 1–10. 10.1371/journal.pntd.0002140Search in Google Scholar PubMed PubMed Central
Silveira M. 2010. Scanning microscopy of ciliary organelles. In: (Eds: A. Mendés-Vilas and J. Díaz) Microscopy: Science, Technology, Applications and Education. Vol. 1. Microscopy Book Series, Formatex Research Center, 156–161Search in Google Scholar
Solaymani-Mohammadi S., Petri Jr W.A. 2006. Zoonotic implications of the swine-transmitted protozoal infections. Veterinary Parasitology, 140, 189–203. https://doi.org/10.1016/j.vetpar.2006.05.01210.1016/j.vetpar.2006.05.012Search in Google Scholar PubMed
Skotarczak B. 1997. Ultrastructural and Cytochemical identification of peroxisomes in Balantidium coli, Ciliophora. Folia biologica (Kraków), 45, 117–120Search in Google Scholar
Skotarczak B. 1999. Cytochemical identification of mucocysts in Balantidium coli trophozoites. Folia biologica (Kraków), 47, 61–65Search in Google Scholar
Van Hoek A.H.A.M, Akhmanova A.S., Huynen M.A., Hackstein H.P. 2000. A mitochondrial ancestry of the hydrogenosomes of Nyctotherus ovalis. Molecular Biology and Evolution, 17, 202–20610.1093/oxfordjournals.molbev.a026234Search in Google Scholar PubMed
Yarllet N., Hann A.C., Lloyd D. 1983. Hydrogenosomes in a mixed isolate of Isotricha prostoma and Isotricha intestinalis from ovine rumen contents. Comparative Biochemistry and Physiology Part B: Comparative Biochemistry. 74, 357–364. https://doi.org/10.1016/0305-0491(83)90025-110.1016/0305-0491(83)90025-1Search in Google Scholar
Zaman V. 1978. Balantidium coli. In: (Ed. J.P. Kreier). Parasitic protozoa. Vol. 2. Academic Press, New York, pp. 633–653Search in Google Scholar
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