• Title/Summary/Keyword: protargol staining

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New record of five Euplotes species(Protozoa, Ciliophora) collected from South Korea

  • Jeong Hyeon Yeo;Pablo Quintela-Alonso;Jae-Ho Jung
    • Journal of Species Research
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    • v.12 no.3
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    • pp.203-211
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    • 2023
  • Five ciliate species of Euplotes were isolated from fresh and coastal water during a sampling survey to identify unrecorded ciliates in South Korea. Their morphology was investigated using live observation, protargol and "wet" silver nitrate staining methods. Brief descriptions and microphotographs of each species and a comparison with related species are provided. Euplotes focardii is characterized by an average size of 65×47 ㎛ after protargol impregnation, 6 dorsal and 3 ventral ridges and dorsal argyrome pattern of double-eurystomus type. Euplotes nobilii shows an average size of 34×20 ㎛ after protargol staining, 6 dorsal and 3 ventral ridges and dorsal argyrome pattern of double-patella type. Euplotes octocarinatus, the only freshwater species described in the present study, is characterized by an average size of 66×46 ㎛ after protargol impregnation, 6 dorsal and 3 ventral ridges and dorsal argyrome pattern of double-patella type. Euplotes petzi has an average size of 43×30 ㎛ after protargol staining, a macronucleus hook-shaped and dorsal argyrome pattern in double-patella type. Euplotes raikovi is characterized by an average size of 40×24 ㎛ after protargol staining, 6 dorsal and 3 ventral ridges and dorsal argyrome pattern of double-patella type.

Single-cell PCR on protargol-impregnated euplotid ciliates: a combined approach of morphological and molecular taxonomy

  • Kim, Se-Joo;Choi, Joong-Ki;Ryu, Seong-Ho;Min, Gi-Sik
    • Animal cells and systems
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    • v.15 no.3
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    • pp.251-258
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    • 2011
  • Ciliates are considered one of the most diverse protozoa and play significant roles in ecology. For successful taxonomic study of these microscopic eukaryotes, a staining procedure is necessary, due mainly to intrinsic difficulties in recognizing characteristics from living cells. Although molecular taxonomy has been used to resolve the ambiguities associated with traditional morphology-based taxonomy, extraction of genomic DNA from stained ciliate cells is not available yet. In the present study, we describe a method to extract genomic DNA from a single protargol-impregnated euplotid cell. By using $HgCl_2$ as a fixative and modulating the exposure time of bleach solution in the protargol impregnation, high-quality genomic DNA can successfully be extracted from a stained single cell with minimal loss of morphological integrity. This technique will contribute to the effectiveness of combined approaches of molecular and morphological taxonomy from single ciliate cells.

New record of three Aspidisca species(Protozoa, Ciliophora) from South Korea

  • Ji Hye Choi;Atef Omar;Jae-Ho Jung
    • Journal of Species Research
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    • v.12 no.1
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    • pp.90-94
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    • 2023
  • The morphology and infraciliature of three newly recorded Aspidisca species in Korea, two collected from the eastern coast and one collected from Jeju Island, were investigated in vivo and after protargol impregnation. The three species are as follows: A. dentata Kahl, 1928, A. hexeris Quennerstedt, 1869, and A. polystyla Stein, 1859. The three species are characterized by having a "polystyla-arrangement" of frontoventral cirri: 1) A. dentata is characterized by having a broadly rotund body shape, a distinct peristomial spur, and a dorsal thorn; 2) A. hexeris is characterized by a broadly oval body shape, four projections along the left margin of body, and the single peristomial spur; and 3) A. polystyla has the broadly rotund body shape, transverse cirri each split into several parts (especially in vivo), and lacking of the peristomial spur. Among them, A. dentata and A. polystyla are poorly known and lack morphological description based on silver staining. In the present study, we provide a brief diagnosis, remarks, and photomicrographs.

First Record of Six Marine Ciliate Species of Genus Strombidium (Ciliophora: Spirotricha: Oligotrichia) from Korea with Ecological Notes

  • Lee, Eun-Sun;Xu, Dapeng;Shin, Mann-Kyoon;Kim, Young-Ok
    • Animal Systematics, Evolution and Diversity
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    • v.28 no.3
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    • pp.192-207
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    • 2012
  • To supply the morphological and ecological information of oligotrich ciliates in Korea, water samples were seasonally collected in Gwangyang Bay and Jinhae Bay from August, 2010 to February, 2012 and processed by quantitative protargol staining method. As a result, six species belonging to the genus Strombidium Clapar$\grave{e}$de and Lachmann, 1859 are identified: Strombidium emergens Kahl, 1932; S. dalum Lynn et al., 1988; S. epidemum Lynn et al., 1988; S. tressum Lynn et al., 1988; S. bilobum Lynn and Gilron, 1993; S. pollostomum Lynn and Gilron, 1993. These six species were newly reported from Korean coastal waters. Strombidium emergens is a middle sized Strombidium ($20-50{\mu}m$ in length) and has open and deep oral groove to girdle portion. Strombidium dalum is a small sized Strombidium (${\geq}20{\mu}m$ in length), that has torch-like spiral anterior membranelles and an inverted triangles-shaped macronucleus on the posterior pole. The small sized S. epidemum has conspicuous trichites surrounding the girdle portion and ventral membranelles distinctly separated from anterior membranelles. The small sized S. tressum has torch-like spiral and extremely long trichites among the cilia of anterior membranelles. The middle sized S. bilobum has a bilobed macronucleus. Strombidium pollostomum is also a small sized Strombidium but its ventral membranelles are continuously connected with anterior membranelles. The five species except S. emergens occurred frequently over the wide range of water temperatures and salinities.

New record of Pleuronema marinum Dujardin, 1841 (Protozoa, Ciliophora) from South Korea

  • Jeong Hyeon, Yeo;Ji Hye, Choi;Atef, Omar;Jae-Ho, Jung
    • Journal of Species Research
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    • v.11 no.4
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    • pp.278-286
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    • 2022
  • During a field survey of Korean marine ciliates, we collected Pleuronema marinum from a brackish water sample. It is characterized by the presence of a contractile vacuole in mid-body, rather than the subterminal/ terminal contractile vacuole as in other congeners. The cells were examined in vivo and based on protargol and 'wet' silver nitrate impregnation. In addition, the nuclear 18S rRNA gene was sequenced using a single cell. The Korean population morphologically and molecularly resembles a Chinese population of P. marinum. Historical review of the species concludes that 1) two or more species have been assigned into P. marinum, 2) the position of contractile vacuole (e.g., in mid-body) is a valid character state, and 3) P. marinum is probably a rare species. Here we provide a monographic treatment of P. marinum to clarify the issue and for further studies of relevant species. Considering there are about 40 nominal species and complex nomenclatural acts in the genus Pleuronema, further studies should provide descriptions based on protargol and 'wet' silver impregnation with marker gene(s).

New Record of Seven Soil Ciliates (Ciliophora: Stichotrichia) from Korea

  • Chae, Kyu-Seok;Kim, Kang-San;Min, Gi-Sik
    • Animal Systematics, Evolution and Diversity
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    • v.37 no.3
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    • pp.212-218
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    • 2021
  • The seven unrecorded ciliates were collected from soil samples in Korea. The morphological study of these ciliates was performed based on observation of both living and stained cells: Rigidocortex quadrinucleatus Bharti, Kumar and La Terza, 2017, Rigidohymena tetracirrata (Gellért, 1942) Berger, 2011, Urosoma salmastra (Dragesco and Dragesco-Kernéis, 1986) Berger, 1999, Urosomoida agilis (Engelmann, 1862) Foissner, 1982, Rigidosticha italiensis Bharti, Kumar and La Terza, 2016, Eschaneustyla terricola Foissner, 1982 and Pseudokeronopsis similis (Stokes, 1886) Borror and Wicklow, 1983. Among the seven genera in this study, the genus Rigidosticha Bharti, Kumar and La Terza, 2016 is firstly reported in Korea. In this study, we provide brief descriptions and remarks for the ciliates with photographs.

New record of three hypotrich soil ciliates(Ciliophora: Hypotricha) from South Korea: Oxytricha multilineata, Mixophrya pantanalensis pantanalensis and Caudiurostyla sinensis

  • Kyu-Seok Chae;Gi-Sik Min
    • Journal of Species Research
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    • v.12 no.4
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    • pp.307-312
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    • 2023
  • Oxytricha multilineata, Mixophrya pantanalensis pantanalensis, and Caudiurostyla sinensis were isolated from soil samples collected from Cheongju-si and Yeoju-si, confirmed as new to South Korea. Oxytricha multilineata was distinguished from other congeners by seven dorsal kineties and dorsal bristles about 15 ㎛ long. Mixophrya pantanalensis pantanalensis was characterized by five to seven lithosomes and six dorsal kineties. Caudiurostyla sinensis was characterized by colorless cortical granules present, 10-14 midventral pairs, 7-9 left and 6-9 right marginal rows and four or five dorsal kineties. We determined the ribosomal DNA sequences (including 18S rDNA, ITS1, 5.8S rDNA, ITS2, and partial 28S rDNA) from above three species. And the genetic distances were compared with their congeners.

Novel Discovery of Two Heterotrichid Ciliates, Climacostomum virens and Fabrea salina (Ciliophora: Heterotrichea: Heterotrichida) in Korea

  • Kim, Ji Hye;Shin, Mann Kyoon
    • Animal Systematics, Evolution and Diversity
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    • v.31 no.3
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    • pp.182-190
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    • 2015
  • Two heterotrichid ciliates, Climacostomum virens (Ehrenberg, 1838) Stein, 1859 from brackish water and freshwater, and Fabrea salina Henneguy, 1890 from a solar saltern, were collected in Korea. They are novelly investigated in Korea by means of live observation, protargol staining and nuclear small subunit (SSU) rRNA gene sequencing. Climacostomum virens is characterized by pouch-like body shape, body length of $200-370{\mu}m$ in vivo, conspicuous cytopharyngeal tube, macronuclei ribbon-like shape, and one to four in number, with or without symbiont algae in cytoplasm, 34-66 somatic kineties, 67-113 adoral zone of membranelles, 8-42 peristomial kineties, 24-37 apical membranelles. SSU rDNA sequence size is 1,591 bp and GC contents 48.52%. Fabrea salina is also characterized by scoop-like body shape with proboscis, body length of $190-240{\mu}m$ in vivo, one to two rod-shaped macronuclei, oval micronuclei, grayish green cortical granules, 104-186 somatic kineties, 4-8 preoral kineties, 7-19 peristomial kineties and fragmented paroral membrane. SSU rDNA sequence size is 1,598 bp and GC contents 47.50%.

Taxonomic Redescription of Loxophyllum perihoplophorum and L. rostratum (Ciliophora: Pleurostomatida) from Korea

  • Kim, Se-Joo;Min, Gi-Sik
    • Animal Systematics, Evolution and Diversity
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    • v.31 no.4
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    • pp.277-283
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    • 2015
  • Two pleurostomatid ciliates, Loxophyllum perihoplophorum Buddenbrock, 1920 and L. rostratum Cohn, 1866, were collected from the coastal waters of the East Sea, Korea. Their morphologies are described based on live observation and protargol staining, and morphometrics are provided. Loxophyllum perihoplophorum is characterized by the following features: 200-650 μm long in vivo; body slender leaf-shaped, flexible and contractile, with thin and wide extrusome-belted zone; 2 macronuclear nodules (Ma) and 1 micronucleus (Mi); 7-9 contractile vacuoles (CV) positioned along dorsal margin; extrusomes (Ex) evenly distributed along edge of entire body, with about 10 dorsal warts (Wa); 9-11 left (LSK) and 19-22 right somatic kineties (RSK), 4-5 furrows (Fu) on left side. Loxophyllum rostratum is about 100-130 μm long in vivo; body oblate leaf-shaped, contractile, convex ventral side and S-shaped dorsal side, beak-like anterior end; 2 Ma and 1 Mi; 1 CV terminally located; Ex distributed along edge of entire body, with about 9-10 dorsal Wa; 7-8 LSK and 15-19 RSK, ca. 5 Fu on left body side. In addition, sequences of small subunit ribosomal DNA were determined from these two Loxophyllum species and compared with the known Loxophyllum sequences.