• ALGAE
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• 제38권2호
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• pp.127-139
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• 2023

Cryptic stages of diverse macroalgae present in natural substrata, "the bank of microscopic forms", were isolated into clonal cultures and identified based on both morphological characteristics and DNA barcoding. Approximately 120 clonal isolates from 308 natural substratum samples were collected from the entire coastline of Kuwait. Amongst these isolates, 77 (64%) were identified through DNA barcoding using the nuclear ribosomal small subunit, RuBisCO spacer (ITS2, tufa, rbcL, psaA, and psbA) and sequencing. Twenty-six isolates (34%) were identified in the division Chlorophyta, 18 (23%) as Phaeophyceae, and 33 (43%) as Rhodophyta. For all DNA sequences in this study, species-level cut off applied was ≥98% homology which depend entirely on the markers used. Three putative new records of Chlorophyta new for the Arabian Gulf were made: Cladophora laetevirens (Dillwyn) Kützing, Ulva torta (Mertens) Trevisan and Ulvella leptochaete (Huber) R. Nielsen, C. J. O'Kelly & B. Wysor in Nielsen, while Cladophora gracilis Kützing and Ulva ohnoi M. Hiraoka & S. Shimada are new records for Kuwait. For Phaeophyceae, Ectocarpus subulatus Kützing and Elachista stellaris Areschoug were new records for the Gulf and Kuwait. In the Rhodophyta, Acrochaetium secundatum (Lyngbye) Nägeli in Nägeli & Cramer, Ceramium affine Setchell & N. L. Gardner, Gelidium pusillum var. pakistanicum Afaq-Husain & Shameel and Dasya caraibica Børgesen are new records for the Gulf and Kuwait, while the red alga Stylonema alsidii (Zanardini) K. Drew is a new record for Kuwait. Several isolates identified corresponded to genera not previously reported in Kuwait and / or the Arabian Gulf, such as Porphyrostromium Trevisan, a new genus from the Bangiales, and two unidentified species for the Planophilaceae Škaloud & Leliaert. The isolates cultivated from substrata enhance understanding of the marine macroalgal diversity in the region and confirmed that the Germling Emergence Method is suitable for determining the actual diversity of a given study area through isolation from cryptic life-history phases.

• Mycobiology
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• 제31권1호
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• pp.19-22
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• 2003

The primary objective of the current research was to detect genetic variations within the Indonesian isolates of Cochliobolus heterostrophus collected from ecologically different places of the country at molecular level using PCR-RFLP analyses. The primer pair of NS3 and NS6 produced amplification fragment in all of the isolates tested. A single fragment of estimated 907 bp was observed in the PCR product pattern. RFLP analysis of the PCR product employing three restriction enzymes, HaeIII, HhaI, and RsaI, respectively, did not reveal intraspecific variations within the fungus. Similarly, nucleotide sequences of portion of small subunit of the ribosomal DNA gene of two of the isolates collected showed no appreciable differences, indicating the absence of genetic diversities among the isolates tested. A phylogenetic tree was constructed and the Indonesian C. heterostrophus, represented by SM-1 isolate, was found to be phylogenetically located near C. sativus, a closely related species.

• Animal Systematics, Evolution and Diversity
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• 제31권4호
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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.

• Parasites, Hosts and Diseases
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• 제52권3호
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• pp.299-304
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• 2014

This study aimed to identify the assemblages (or subassemblages) of Giardia duodenalis by using normal or nested PCR based on 4 genetic loci: glutamate dehydrogenase (gdh), triose phosphate isomerase (tpi), ${\beta}$-giardin (bg), and small subunit ribosomal DNA (18S rRNA) genes. For this work, a total of 216 dogs' fecal samples were collected in Guangdong, China. The phylogenetic trees were constructed with MEGA5.2 by using the neighbor-joining method. Results showed that 9.7% (21/216) samples were found to be positive; moreover, 10 samples were single infection (7 isolates assemblage A, 2 isolates assemblage C, and 1 isolate assemblage D) and 11 samples were mixed infections where assemblage A was predominant, which was potentially zoonotic. These findings showed that most of the dogs in Guangdong were infected or mixed-infected with assemblage A, and multi-locus sequence typing could be the best selection for the genotype analysis of dog-derived Giardia isolates.

• Mycobiology
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• 제42권2호
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• pp.174-180
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• 2014

We analyzed the genetic diversity of Cylindrocarpon destructans isolates obtained from Korean ginseng (i.e., Panax ginseng) roots by performing virulence tests and nuclear ribosomal gene internal transcribed spacer (ITS) and mitochondrial small subunit (mt SSU) rDNA sequence analysis. The phylogenetic relationship analysis performed using ITS DNA sequences and isolates from other hosts helped confirm that all the Korean C. destructans isolates belonged to Nectria/Neonectria radicicola complex. The results of in vivo and ex vivo virulence tests showed that the C. destructans isolates could be divided into two groups according to their distinctive difference in virulence and the genetic diversity. The highly virulent Korean isolates in pathogenicity group II (PG II), together with foreign isolates from P. ginseng and P. quinquefolius, formed a single group. The weakly virulent isolates in pathogenicity group I, together with the foreign isolates from other host plants, formed another group and exhibited a greater genetic diversity than the isolates of PG II, as confirmed by the mt SSU rDNA sequence analysis. In addition, as the weakly virulent Korean isolates were genetically very similar to the foreign isolates from other hosts, they were likely to originate from hosts other than the ginseng plants.

• ALGAE
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• 제37권4호
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• pp.249-264
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• 2022

Two genera of the Rhodymeniales, Halopeltis and Leptofauchea, are here reported for the first time from the Hawaiian Islands and represent the deepest records for both genera. Molecular phylogenetic analyses of cytochrome oxidase subunit I (COI), rbcL, and large subunit ribosomal DNA (LSU) sequences for Hawaiian specimens of Leptofauchea revealed one well-supported clade of Hawaiian specimens and three additional lineages. One of these clades is described here as Leptofauchea huawelau sp. nov., and is thus far known only from mesophotic depths at Penguin Bank in the Main Hawaiian Islands. L. huawelau sp. nov. is up to 21 cm, and is the largest known species. An additional lineage identified in the LSU and rbcL analyses corresponds to the recently described L. lucida from Western Australia, and is a new record for Hawai'i. Hawaiian Halopeltis formed a well-supported clade along with H. adnata from Korea, the recently described H. tanakae from mesophotic depths in Japan, and H. willisii from North Carolina, and is here described as Halopeltis nuahilihilia sp. nov. H. nuahilihilia sp. nov. has a distinctive morphology of narrow vegetative axes that harbor constrictions along their length. The current distribution of H. nuahilihilia includes mesophotic depths around W. Maui, W. Moloka'i, and the island of Hawai'i in the Main Hawaiian Islands. Few reproductive characters were observed because of the small number of specimens available; however, both species are distinct based on phylogeny and morphology. These descriptions further emphasize the Hawaiian mesophotic zone as a location harboring many undescribed species of marine macroalgae.

• Journal of Microbiology and Biotechnology
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• 제15권5호
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• pp.1028-1038
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• 2005

To elucidate phylogenetic relationships of Phellinus and its related genera, nuclear internal transcribed spacer and mitochondrial small subunit ribosomal DNA sequences from 65 strains were determined and compared. The combined dataset of two sequences increased informative characters and led to the production of trees with higher levels of resolution. Phylogenetic analysis of the combined dataset revealed thirteen evolutionary lineages and several unresolved species that were together subdivided into two large clusters consisting of oligonucleate species and binucleate species. These results coincided with previous cytological, morphological, and molecular studies. It is newly recognized that the Phellinus linteus complex forms a sister clade to Inonotus, and that Fulvifomes is somehow related to Inocutis. The Phellinus linteus complex of dimitic perennial taxa made an independent clade from Inonotus and suggested that hyphal miticity and fruitbody permanence had enough phylogenetic significance to keep the complex within the traditional genus Phellinus. Taxa lacking setae were clustered into Fulvifomes, Phylloporia, Inocutis, and Fomitiporia, and the first three were closely related sister groups, but Fomitiporia was a genus distantly related to them. Several taxa with branched setae were shown among distantly related genera. Molecular evidence indicated that the ancestral nuclear type could be a binucleate feature, and that there might be parallel gains of branched setae and parallel losses of setae in the Hymenochaetales.

• Animal Systematics, Evolution and Diversity
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• 제27권1호
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• pp.25-33
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• 2011

Three morphospecies of the genus Uronychia, i.e. U. setigera Calkins, 1902, U. binucleata Young, 1922, and U. multicirrus Song, 1997, were collected from the coastal waters of Gumjin-ri on the East Sea and the public waterfront of Incheon on the Yellow Sea in Korea, respectively. These species are described based on live observation, protargol impregnation, silver nitrate impregnation, and their morphometrics. Diagnostic keys for these species are also provided. In addition, their small subunit ribosomal DNA sequences were compared with previously known sequences of Uronychia species. Diagnostics of three Uronychia species are as follows: U. setigera: $50-80\;{\mu}m$ long in vivo, oval-shaped, 2 macronuclear nodules (Ma), 1 spur on the left margin, 11 adoral membranelles (AM) 1, 4 AM2, 1 buccal cirrus (BC), 4 frontal cirri (FC), 3 left marginal cirri (LMC), 2 ventral cirri (VC), 5 transverse cirri (TC), 3 caudal cirri (CC), 6 dorsal kineties (DK), and approximately 23 cilia in the leftmost kinety. U. binucleata: $70-110\;{\mu}m$ long in vivo, oval to slightly rectangular shaped, 2 Ma, 1 micronucleus (Mi), 2 spurs on the posterior region, 11 AM1, 4 AM2, 1 BC, 4 FC, 3 LMC, 2 VC, 5 TC, 3 CC, 6 DK, and approximately 37 cilia in the leftmost kinety. U. multicirrus: $140-200\;{\mu}m$ long in vivo, oval to slightly rectangular shaped, ca. 7 Ma, 1 Mi, 11 AM1, 4 AM2, 1 BC, 4 FC, 3 LMC, approximately 8 VC, 5 TC, 3 CC, and 6 DK. This study presents the first record of this genus in Korea.

• Parasites, Hosts and Diseases
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• 제42권3호
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• pp.93-119
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• 2004

Acanthamoeba and Naegleria are widely distributed in fresh water, soil and dust throughout the world, and cause meningoencephalitis or keratoconjunctivitis in humans and other mammals. Korean isolates, namely, Naegleria sp. YM-1 and Acanthamoeba sp. YM-2, YM-3, YM-4, YM-5, YM-6 and YM-7, were collected from sewage, water puddles, a storage reservoir, the gills of a fresh water fish, and by corneal washing. These isolates were categorized into three groups based on the mortalities of infected mice namely, highly virulent (YM-4), moderately virulent (YM-2, YM-5 and YM-7) and nonpathogenic (YM-3). In addition, a new species of Acanthamoeba was isolated from a freshwater fish in Korea and tentatively named Korean isolate YM-4. The morphologic characters of its cysts were similar to those of A. culbertsoni and A. royreba, which were previously designated as Acanthamoeba group III. Based on experimentally infected mouse mortality, Acanthamoeba YM-4 was highly virulent. The isoenzymes profile of Acanthamoeba YM-4 was similar to that of A. royreba. Moreover, an anti-Acanthamoeba YM-4 monoclonal anti-body reacted only with Acanthamoeba YM-4, and not with A. culbertsoni. Random amplified polymorphic DNA marker analysis and RFLP analysis of mitochondrial DNA and of a 188 small subunit ribosomal RNA, placed Acanthamoeba YM-4 in a separate cluster based on phylogenic distances. Thus Acanthamoeba YM-4 was identified as a new species, and assigned Acanthamoeba sohi. Up to the year 2002 in Korea, two clinical cases were found to be infected with Acanthamoeba spp. These patients died of meningoencephalitis. In addition, one case of Acanthamoeba pneumonia with an immunodeficient status was reported and Acanthamoeba was detected in several cases of chronic relapsing corneal ulcer, chronic conjunctivitis, and keratitis.

• Parasites, Hosts and Diseases
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• 제39권2호
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• pp.161-170
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• 2001

We conducted both the small subunit ribosomal DNA (SSU rDNA) polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and mitochondrial (mt) DNA RFLP analyses for a genetic characterization of Acanthamoeba isolates from contact lens storage cases of students in Seoul, Korea. Twenty-three strains of Acanthamoeba from the American Type Culture Collection and twelve clinical isolates from Korean patients were used as reference strains. Thirty-nine isolates from contact lens storage cases were classified into seven types (KA/LS1 , KA/LS2, KA/LS4, KA/LS5, KA/LS7 KA/LS18, KA/LS31). Four types (KA/LS1 , KA/LS2, KA/LS5, KA/LS18) including 33 isolates were regarded as A. castellanii complex by riboprints. KA/LS1 type was the most predominant (51.3%) in the present survey area, followed by KA/LS2 (20.9%), and KA/LSS (7.7%) types. Amoebae of KA/LS1 type had the same mtDNA RFLP and riboprint patterns as KA/E2 and KA/E12 strains, clinical isolates from Korean keratitis patients. Amoebae of KA/LS2 type had the identical mtDNA RFLP patterns with A. castellanii Ma strain, a corneal isolate from an American patient as amoebae of KA/LS5 type, with KA/E3 and KA/E8 strains from other Korean keratitis patients. Amoebae of KA/LS 18 type had identical patterns with JAC/E1, an ocular isolate from a Japanese patient. Three types , which remain unidentified at species level, were not corresponded with any clinical isolate in their mtDNA RFLP and riboprint patterns. Out of 39 isolates analyzed in this study, mtDNA RFLP and riboprint patterns of 33 isolates (84.6%) were identical to already known clinical isolates, and therefore, they may be regarded as potentially keratopathogenic. These results suggest that contact lens wearers in Seoul should pay more attention to hygienic maintenance of contact lens storage cases for the prevention of Acanthamoeba keratitis.