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Original Article

Morphological and Molecular Characteristics of Clinostomid Metacercariae from Korea and Myanmar

Eun Jeong Won1, Yu Jeong Lee1, Moon-Ju Kim1, Jong-Yil Chai2,3orcid, Byoung-Kuk Na4orcid, Woon-Mok Sohn4,*orcid
The Korean Journal of Parasitology 2020;58(6):635-645.
Published online: December 31, 2020

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*Corresponding author (wmsohn@gnu.ac.kr)
• Received: August 31, 2020   • Revised: October 18, 2020   • Accepted: November 13, 2020

Copyright © 2020 by The Korean Society for Parasitology and Tropical Medicine

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Morphological and Molecular Characteristics of Clinostomid Metacercariae from Korea and Myanmar
Korean J Parasitol. 2020;58(6):635-645.   Published online December 29, 2020
DOI: https://doi.org/10.3347/kjp.2020.58.6.635
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Morphological and Molecular Characteristics of Clinostomid Metacercariae from Korea and Myanmar
Korean J Parasitol. 2020;58(6):635-645.   Published online December 29, 2020
DOI: https://doi.org/10.3347/kjp.2020.58.6.635
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Morphological and Molecular Characteristics of Clinostomid Metacercariae from Korea and Myanmar
Image Image Image Image Image
Fig. 1 Light microscopic findings of clinostomid metacercariae (CMc) collected in fish from Korea and Myanmar. (A) Type A CMc collected from Squalidus japonicus coreanus in Yang-cheon, Sancheong-gun, Gyeongsangnam-do. (B) Type B CMc in Carassius auratus from Nakdong-gang in Sangju-si, Gyeongsangbuk-do. (C) Type C CMc in C. auratus from Songji-ho in Goseong-gun, Gangwon-do. (D) CMc detected in snakehead, Channa sp., from Yangon, Myanmar. Scale bar is 1 mm.
Fig. 2 Intestinal ceca of clinostomid metacercariae (CMc). (A) Korean CMc with smooth thin wall and folded lumen. (B) Myanmar CMc with relatively thick wall and bilaterally branched lumen. Scale bar=200 μm.
Fig. 3 Scanning electron micrographs of Korean clinostomid metacercariae collected in Carassius auratus from Nakdong-gang in Sangju-si, Gyeongsangbuk-do. (A) Whole ventral view. (B) Whole dorsal view. (C) Ventral sucker with numerous dome-shaped sensory papillae. (D) Tegumental surface on the ventro-median portion with no papillae-like spinous processes (square in Fig. 3A). (E) Tegumental surface on the dorso-median portion with numerous small spinous processes (square in Fig. 3B). (F) Tegumental surface on the dorso-posterior portion with numerous small spinous processes (encircled in Fig. 3B).
Fig. 4 Scanning electron micrographs of Myanmar clinostomid metacercariae collected in a snakehead, Channa sp., from Yangon, Myanmar. (A) Whole ventral view (Scale bar=1,000 μm). (B) Whole dorsal view (Scale bar=1,000 μm). (C) Oral sucker (Scale bar=200 μm). (D) Ventral sucker (Scale bar=100 μm). (E) Tegumental surface around the genital pore (encircled in Fig. 4A) in 2/5 posterior portion of the body (Scale bar=50 μm). (F) Tegument around the Laurer’s canal (encircled in Fig. 4B) at 1/4 dorso-posterior portion of the body. (G) Tegument between the oral and ventral suckers with no papillae-like spinous processes (squares in Fig. 4A). (H) Tegumental surface on the dorso-middle portion with numerous papillae-like spinous processes, which were broadly distributed on the body surface except for anterior portion (squares in Fig. 4A,B) (Scale bar=25 μm).
Fig. 5 Phylogenetic tree constructed based on CO1 sequences from flukes collected in this study and from platyhelminth sequences deposited in GenBank. The bootstrap values for the clade with consensus support (%) were displayed on the tree to the left of the most recent common ancestral node for that clade. The scale bar at the bottom of the tree indicated the length of the branches of the tree.

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Fig. 5

Morphological and Molecular Characteristics of Clinostomid Metacercariae from Korea and Myanmar

Comparative dimensionsa of clinostomid metacercariae (CMc) in fish from Korea and Myanmar (unit: μm)

Organs Korean CMcb Myanmar CMcc
Type A Type B Type C
Body length (BL) 5,300–7,725 (6,158) 3,700–7,600 (5,728) 4,375–5,850 (4,958) 6,550–8,950 (7,675)
Body width (BW) 1,400–1,875 (1,675) 1,100–1,975 (1,510) 2,000–2,300 (2,203) 2,125–3,375 (2,778)
Ratio of BL/BW 2.97–4.80 (3.73) 2.53–4.66 (3.89) 1.97–2.72 (2.26) 2.46–3.12 (2.78)
Oral sucker (OSL) 320–400 (360) 270–350 (314) 280–380 (311) 380–500 (443)
Oral sucker (OSW) 360–500 (433) 320–420 (381) 360–460 (407) 500–570 (543)
Ventral sucker (VSL) 760–950 (901) 740–990 (806) 810–980 (908) 1,030–1,400 (1,232)
Ventral sucker (VSW) 760–960 (900) 740–930 (794) 770–990 (900) 1,020–1,400 (1,200)
VSW/OSW 1.70–2.29 (2.09) 1.88–2.31 (2.06) 1.89–2.43 (2.22) 1.92–2.50 (2.21)
Distance between suckers 480–900 (652) 350–780 (599) 370–570 (458) 550–800 (655)
VSW/BW 0.47–0.64 (0.54) 0.42–0.67 (0.54) 0.36–0.50 (0.41) 0.40–0.51 (0.43)

a(L): length; (W): width; unit is μm.

bType A: collected in Squalidus japonicus koreanus from Yang-cheon in Sancheong-gun, Gyeongsangnam-do; Type B: in Carassius auratus from Nakdong-gang in Sangju-si, Gyeongsangbuk-do; Type C: in C. auratus from Songji-ho in Goseong-gun, Gangwon-do.

cIn Channa sp. from Yangon, Myanmar.

Matched base pairs (percent) of the CO1 sequences in specimens from the present and previous studies

Matched base pairs (%) to the sequences of
Species Clinostomum complanatum Clinostomum phalacrocoracis Clinostomum brieni Fasciola gigantica
Accession No. KM923964 MH491530 JF718595 JF718588 KY906238 KY906236 MH253048 MH253047 MF287791 KF543342
Country China Japan Italy Italy South Africa South Africa Congo Congo Viet Nam Viet Nam
Isolate No. 1 (Korea) 736/736 (100.0) 353/353 (100.0) 282/296 (95.3) 281/296 (94.9) 251/285 (88.1) 251/285 (88.1) 249/284 (87.7) 249/284 (87.7) 597/749 (79.7) 595/749 (79.4)
Isolate No. 2 (Korea) 728/728 (100.0) 344/344 (100.0) 282/295 (95.6) 281/296 (94.9) 251/285 (88.1) 251/285 (88.1) 249/284 (87.7) 249/284 (87.7) 588/741 (79.4) 587/741 (79.2)
Isolate No. 3 (Korea) 727/728 (99.9) 344/344 (100.0) 282/295 (95.6) 280/296 (94.6) 250/285 (87.7) 250/285 (87.7) 248/284 (87.3) 248/284 (87.3) 590/741 (79.6) 588/741 (79.4%)
Isolate No. 4 (Korea) 733/734 (99.9) 351/351 (100.0) 281/296 (94.9) 280/296 (94.6) 250/285 (87.7) 250/285 (87.7) NA NA NA NA
Isolate No. 5 (Korea) 731/734 (99.6) 363/366 (99.2) 268/282 (95.0) 267/282 (94.7) 239/270 (88.5) 239/270 (88.5) NA NA 575/714 (80.5) 570/714 (79.8)
Isolate No. 6 (Myanmar) 777/884 (87.9) NA 532/616 (86.4) NA 550/621 (88.6) 550/621 (88.6) 541/620 (87.3) NA NA NA

Matched base pairs (percent) of the ITS1 sequences in specimens from the present and previous studies

Matched base pairs (%) to the sequences of
Species Clinostomum complanatum Clinostomum cutaneum Clinostomum phalacrocoracis Euclinostomum heterostomum
Accession no. FJ609420 AY245701 JF718629 MH845236 GQ339114 FJ609421 FJ609422 FJ609423 KY312847 KY273278
Country Italy Israel Italy Iran Kenya Kenya Kenya Kenya India India
Isolate No. 1 (Korea) 694/695 (99.9) 694/695 (99.9) 694/695 (99.9) 659/660 (99.8) 680/695 (97.8) 680/695 (97.8) 680/695 (97.8) 680/695 (97.8) NA NA
Isolate No. 2 (Korea) 693/694 (99.9) 693/694 (99.9) 664/665 (99.8) 659/660 (99.8) 679/694 (97.8) 679/694 (97.8) 679/694 (97.8) 679/694 (97.8) NA NA
Isolate No. 3 (Korea) 697/698 (99.9) 697/698 (99.9) 697/698 (99.9) 662/663 (99.8) 683/698 (97.9) 683/698 (97.9) 683/698 (97.9) 683/698 (97.9) NA NA
Isolate No. 4 (Korea) 693/694 (99.9) 693/694 (99.9) 664/665 (99.8) 659/660 (99.8) 679/694 (97.8) 679/694 (97.8) 679/694 (97.8) 679/694 (97.8) NA NA
Isolate No. 5 (Korea) 691/692 (99.9) 691/692 (99.9) 662/663 (99.8) 657/658 (99.8) 677/692 (97.8) 677/692 (97.8) 677/692 (97.8) 677/692 (97.8) NA NA
Isolate No. 6 (Myanmar) 681/694 (98.1) 681/694 (98.1) NA NA 679/694 (97.8) 679/694 (97.8) 677/694 (97.7) 677/694 (97.7) 668/669 (99.9) 666/669 (99.6)
Table 1 Comparative dimensionsa of clinostomid metacercariae (CMc) in fish from Korea and Myanmar (unit: μm)

(L): length; (W): width; unit is μm.

Type A: collected in Squalidus japonicus koreanus from Yang-cheon in Sancheong-gun, Gyeongsangnam-do; Type B: in Carassius auratus from Nakdong-gang in Sangju-si, Gyeongsangbuk-do; Type C: in C. auratus from Songji-ho in Goseong-gun, Gangwon-do.

In Channa sp. from Yangon, Myanmar.

Table 2 Matched base pairs (percent) of the CO1 sequences in specimens from the present and previous studies
Table 3 Matched base pairs (percent) of the ITS1 sequences in specimens from the present and previous studies

Table 1

Table 2

Table 3

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