• Journal of fish pathology
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• v.17 no.2
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• pp.145-149
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• 2004

The definitive identification of ciliate species by morphological characteristics relies on time-consuming and laborious staining techniques. Therefore, in this study, we discriminated 3 scuticociliatosis causing species - Pseudocohnilembus persalinus, Uronema marinum and Philasterides dicentrarchi - in cultured olive flounder by multiplex PCR. The multiplex PCR based on the species-specific amplification of small subunit ribosomal RNA (SS rRNA) gene sequence enabled us to distinguish the 3 scuticociliate species in a simple and rapid manner, even in the sample containing the three species simultaneously. These data suggest that the multiplex PCR strategy would make it possible to avoid the cumbersome and time-consuming procedures of morphological analysis for the definitive identification of scuticociliates.

• Journal of Species Research
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• v.10 no.1
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• pp.63-71
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• 2021

Anteholosticha bergeri and Bakuella granulifera were isolated from soil samples collected from Muuidong and Songdo-dong, Incheon and confirmed new to South Korea. Including these two newly recorded species, 11 species of Anteholosticha and four species of Bakuella have been recorded in South Korea to date. Anteholosticha bergeri was discriminated from congeners by following characters: cortical granules, 12-16 macronuclei, 5-8 midventral pairs, 2-3 pretransverse cirri, 4-6 transverse cirri, and three dorsal kineties. Bakuella granulifera was identified by cortical granules, 5-11 buccal cirri, 2-5 frontoterminal cirri, 2-5 midventral cirri rows, and 8-12 transverse cirri. The Korean A. bergeri population corresponds to the Austrian population, except for the number of marginal and transverse cirri, and the Korean B. granulifera population corresponds to the Namibian population, except for body size. In addition, small subunit ribosomal RNA(18S rRNA) gene sequences from both species were determined.

• Parasites, Hosts and Diseases
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• v.37 no.3
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• pp.181-188
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• 1999

We investigated the value of mitochondrial small subunit rRNA gene (mt SSU rDNA) PCR-RFLP as a taxonomic tool for Acanthamoeba isolates with close inter-relationships. Twenty-five isolates representing 20 species were included in the analysis. As in nuclear 18s rDNA analysis, two type strains (A. astronyxis and A. tubiashi) of morphological group 1 diverged earliest from the other strains, but the divergence between them was less than in 18s riboprinting. Acanthamoeba griffini of morhological group 2 branched between pathogenic (A. culbertsoni A-1 and A. healyi OC-3A) and nonpathogenic (A.palestinensis Reich, A. pustulosa GE-3a, A. royreba Oak Ridge, and A lenticulata PD2S) strains of morphological group 3. Among the remaining isolates of morphological group 2, the Chang strain had the identical mitochondrial riboprints as the type strain of A. hatchetti. AA2 and AA1, the type strains of A. divionensis and A. paradivionensis, respectively, had the identical riboprints as A. quina Vil3 and A. castellanii Ma. Although the branching orders of A. castellanii Neff, A. polyphaga P23, A. triangularis SH621, and A. lugdunensis L3a were different from those in 18S riboprinting analysis, the results obtained from this study generally coincided well with those from 18S riboprinting. Mitochondrial riboprinting may have an advantage over nuclear 18S rDNA riboprinting beacuse the mt SSU rDNAs do not seem to have introns that are found in the 18S genes of Acanthamoeba and that distort phylogenetic analyses.

• ALGAE
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• v.32 no.3
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• pp.181-187
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• 2017

During a sampling at Nokdong harbor, southern coast of Korea in January 2017, the marine diatom Coscinodiscus wailesii cells infected by a novel parasitoid nanoflagellate were observed. While the development process of the trophosomes of the parasitoid was more similar to that of Pseudopirsonia mucosa, division pattern of the auxosomes was similar to that of Pirsonia species. Phylogenetic analyses inferred from 18S rRNA gene sequences revealed that the parasitoid infecting C. wailesii fell within the cercozoan groups and branched as a sister lineage of the clade consisting of Pseudopirsonia mucosa and the undescribed Cercomonas sp. SIC7235, with the sequence dissimilarity of 7.3% with Pseudopirsonia mucosa. All of these developmental and molecular characteristics suggest that the parasitoid nanoflagellate infecting the diatom C. wailesii is a new Pseudopirsonia species.

• Animal Systematics, Evolution and Diversity
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• v.38 no.4
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• pp.226-237
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• 2022

Arcuospathidium is a haptorian ciliate genus composed of 18 species, and only one species has been reported in Korea. Here, we identify two unrecorded Arcuospathidium subspecies by morphological observation of both living and protargol-impregnated specimens with the small subunit ribosomal RNA (18S rRNA) gene sequence. These subspecies, Arcuospathidium cultriforme cultriforme (Penard, 1922) Foissner, 1984 and A. cultriforme scalpriforme (Kahl, 1930) Foissner, 2003, were isolated from various terrestrial habitats in July and August 2013, respectivley. Arcuospathidium cultriforme cultriforme is similar to A. cultriforme scalpriforme by a knife-shaped body, a twisted-shaped macronucleus, number of dorsal brushes, position of dorsal brushes, and shape of macronucleus but former mainly differs from the body length to oral bulge length ratio (27-38% vs. 41-53%), extrusome (one types vs. three types), cyst shape (roughly faceted wall vs. smooth surface and thin wall) and number of somatic kinety rows(18-30 vs. 30-44). Additionally, we analyzed the 18S rRNA gene sequences of two A. cultriforme subspecies and compared them with the sequences from GenBank to confirm their identification at the molecular level. As the results of genetic analysis, the 18S rRNA gene sequence of the Korean A. cultriforme cultriforme population is most similar to that of Austrian population. Also, the sequence of the Korean A. cultriforme scalpriforme population is most similar to that of another population with some nucleotide differences.

• Parasites, Hosts and Diseases
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• v.47 no.3
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• pp.293-297
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• 2009

Improved methods for detection of Cryptosporidium oocysts in environmental and clinical samples are urgently needed to improve detection of cryptosporidiosis. We compared the sensitivity of 7 PCR primer sets for detection of Cryptosporidium parvum. Each target gene was amplified by PCR or nested PCR with serially diluted DNA extracted from purified C. parvum oocysts. The target genes included Cryptosporidium oocyst wall protein (COWP), small subunit ribosomal RNA (SSU rRNA), and random amplified polymorphic DNA. The detection limit of the PCR method ranged from $10^3$ to $10^4$ oocysts, and the nested PCR method was able to detect $10^0$ to $10^2$ oocysts. A second-round amplification of target genes showed that the nested primer set specific for the COWP gene proved to be the most sensitive one compared to the other primer sets tested in this study and would therefore be useful for the detection of C. parvum.

• Animal Systematics, Evolution and Diversity
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• v.29 no.2
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• pp.144-151
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• 2013

Two marine hypotrichous ciliates, Anteholosticha petzi and Ponturostyla enigmatica, were collected from the Yellow Sea and the Korea Strait, respectively, and described using live observation and protargol-impregnated specimens. Furthermore, the nuclear small subunit ribosomal RNA gene of each was sequenced and compared to previously annotated sequences retrieved from the GenBank. Anteholosticha petzi is characterized by 3 frontal cirri (FC), 2 frontoterminal cirri (FTC), 8-12 transverse cirri (TC), 1 buccal cirrus (BC), 9-12 midventral pairs (MP), 3 bipolar dorsal kineties (DK), and 3 types of colorless cortical granules. Ponturostyla enigmatica is characterized by 8 FC, 5 ventral cirri (VC), 5-7 TC, 6-7 marginal rows (MR) on each side, 4 complete and 2-3 partial DK, and greenish cortical granules. This is the first identification and description of these 2 species, A. petzi and P. enigmatica, in South Korea.

• Parasites, Hosts and Diseases
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• v.53 no.1
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• pp.113-117
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• 2015

Cryptosporidium spp., ubiquitous enteric parasitic protozoa of vertebrates, recently emerged as an important cause of economic loss and zoonosis. The present study aimed to determine the distribution and species of Cryptosporidium in post-weaned and adult pigs in Shaanxi province, northwestern China. A total of 1,337 fresh fecal samples of post-weaned and adult pigs were collected by sterile disposable gloves from 8 areas of Shaanxi province. The samples were examined by Sheather's sugar flotation technique and microscopy at${\times}400$ magnification for Cryptosporidium infection, and the species in positive samples was further identified by PCR amplification of the small subunit (SSU) rRNA gene. A total of 44 fecal samples were successfully amplified by the nested PCR of the partial SSU rRNA, with overall prevalence of 3.3%. The average prevalence of Cryptosporidium infection in each pig farms ranged from 0 to 14.4%. Species identification by sequencing of SSU rRNA gene revealed that 42 (3.1%) samples were Cryptosporidium suis and 2 (0.15%) were Cryptosporidium scrofarum. C. suis had the highest prevalence (7.5%) in growers and the lowest in breeding pigs (0.97%). C. suis was the predominant species in pre-weaned and adult pigs, while C. scrofarum infected pigs older than 3 months only. A season-related difference of C. suis was observed in this study, with the highest prevalence in autumn (5.5%) and the lowest (1.7%) in winter. The present study provided basic information for control of Cryptosporidium infection in pigs and assessment of zoonotic transmission of pigs in Shaanxi province, China.

• Animal Systematics, Evolution and Diversity
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• v.27 no.3
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• pp.220-227
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• 2011

The morphology of the two marine urostyloid ciliates, Pseudokeronopsis carnea (Cohn, 1866) and Uroleptopsis citrina Kahl, 1932, in the family Pseudokeronopsidae, collected from the Yellow Sea, and the East Sea, Korea, respectively, were studied using live observation and protargol impregnation. Additionally, the small subunit ribosomal RNA (SSU rRNA) gene was sequenced. These two species are firstly recorded in Korea. The main diagnostic key is as follows. Pseudokeronopsis carnea: body outline elongate-elliptical, brown-reddish or orange-red in colour in vivo; bicorona of 16-24 frontal cirri; one buccal and two frontoterminal cirri; 7-10 transverse cirri; 5-7 dorsal kineties; two types of cortical granules (one orange-red pigment, mainly grouped around cirri and dorsal bristles, arranged in typical rubra-pattern; the other, colourless and blood-cell-shaped, and densely distributed); contractile vacuole in the posterior half of the cell on the left side, usually in posterior 1/3-2/5. Uroleptopsis citrina: body outline elongate-elliptical, lemon-yellow in colour in vivo; two types of cortical granules (one yellow pigment; the other, blood-cell-shaped, densely distributed); bicorona of 12-18 frontal cirri; 2-3 frontoterminal cirri; two midventral rows comprising 26-35 cirri (consisting of anterior paired cirri, non-paired single cirri, and posterior paired cirri); three dorsal kineties. In addition, the SSU rRNA sequences of the two species were compared with public database of these species and consequently, showed high similarity.

• Parasites, Hosts and Diseases
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• v.53 no.2
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• pp.237-242
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• 2015

Analysis of ancient DNA (aDNA) extracted from Ascaris is very important for understanding the phylogenetic lineage of the parasite species. When aDNAs obtained from a Joseon tomb (SN2-19-1) coprolite in which Ascaris eggs were identified were amplified with primers for cytochrome b (cyt b) and 18S small subunit ribosomal RNA (18S rRNA) gene, the outcome exhibited Ascaris specific amplicon bands. By cloning, sequencing, and analysis of the amplified DNA, we obtained information valuable for comprehending genetic lineage of Ascaris prevalent among pre-modern Joseon peoples.