• 한국어병학회지
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• 제27권1호
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• pp.25-33
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• 2014

Anisakis simplex sensu stricto와 A. pegreffii의 3기 유충과 4기 유충에서 얻은 excretory-secretory (ES) products 및 somatic extracts의 가수분해효소 활성을 API ZYM kit를 이용하여 비교하였다. Esterase 그룹의 가수분해효소 중 acid phosphatase는 A. simplex (s.s.)와 A. pegreffii 모두에서 높은 활성을 나타냈으며, esterase (C 4)의 경우 somatic extracts에서만 가수분해효소 활성이 나타났는데 A. simplex (s.s.)가 A. pegreffii와 비교하여 3기와 4기 유충 모두에서 2배 가량 높은 활성을 나타냈다. alkaline phosphatase, acid phosphatase 그리고 naphthol-AS-BI-phosphohydrolase의 경우 A. simplex (s.s.)와 A. pegreffii 모두 3기 유충보다 4기 유충에서 더 높은 가수분해효소 활성이 확인되었다. Aminopeptidase 그룹의 가수분해효소 활성은 leucine arylamidase에서 관찰되었는데, somatic extracts의 경우 A. pegreffii 보다 A. simplex (s.s.)에서 가수분해 효소 활성이 두 배 가량 높게 확인되었으며, 대부분의 다른 효소들에서는 활성이 거의 나타나지 않았다. Glycosidase 그룹의 가수분해효소 활성은 N-acetyl-${\beta}$-glucosaminidase, ${\alpha}$-mannosidase 그리고 ${\alpha}$-fucosidase에서 확인되었는데, A. simplex (s.s.) 보다 A. pegreffii에서 높은 가수분해효소 활성을 확인할 수 있었으며, 대부분 4기 유충보다 3기 유충에서 더 높은 가수분해효소 활성이 확인되었다. 이러한 아니사키스속 선충의 종과 유충단계에 따른 가수분해 효소의 활성 차이는 선충의 성장, 탈피, 소화, 섭이 등의 대사과정의 차이에 기인한 것으로 생각된다.

• Parasites, Hosts and Diseases
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• 제61권3호
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• pp.292-297
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• 2023

Extra-gastrointestinal anisakidosis is rare. We herein report an Anisakis pegreffii infection in a patient with hepatic anisakidosis diagnosed based on its molecular identification. A 71-year-old male patient had a hepatic tumor presenting as a low-density area of 20 mm in diameter in segment 6 of the liver on abdominal ultrasonography, computed tomography, and magnetic resonance imaging. The surgically resected pathological specimen revealed a necrotizing eosinophilic granuloma containing nematode larvae, possibly an Anisakis larva. Molecular and phylogenetic analysis demonstrated Anisakis larvae belonging to A. pegreffii. The present results will help identify and characterize unknown Anisakis species in histological sections.

• Parasites, Hosts and Diseases
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• 제53권3호
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• pp.349-353
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• 2015

Anisakis simplex sensu stricto (s.s.), Anisakis pegreffii, Anisakis berlandi (=A. simplex sp. C), and Anisakis typica are the 4 major species of Anisakis type I larvae. In the Republic of Korea (Korea), A. pegreffii, A. berlandi, and A. typica larvae in fish hosts has seldom been documented. In this study, molecular analysis was performed on Anisakis larvae from the sea eels (Astroconger myriaster), the major source of human anisakiasis in Korea, collected from Tongyeong City, a southern coastal area of Korea. All 20 sea eels examined were infected with Anisakis type I larvae (160 larvae; 8 per fish). Their species were analyzed using PCR-RFLP patterns and nucleotide sequences of internal transcribed spacers (ITS1, 5.8 subunit gene, and ITS2) and mitochondrial cytochrome c oxidase 2 (cox2). Most (86.8%; 112/129) of the Anisakis type I larvae were A. pegreffii, and 7.8% (10/129) were A. typica. The remaining 5.4% (7/129) was not identified. Thus, A. pegreffii is the major species of anisakid larvae in sea eels of the southern coast of Korea.

• Parasites, Hosts and Diseases
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• 제57권2호
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• pp.207-211
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• 2019

Anisakiasis is a zoonotic disease induced by anisakid nematodes, and endoscopic inspection is used for a diagnosis or remedy for it. Anisakis simplex, Anisakis physeteris, and Pseudoterranova decipiens had been reported to be the major species causing human infections, particularly, in Japan. However, in Korea, recent studies strongly suggested that Anisakis pegreffii is the major species of human infections. To support this suggestion, we collected anisakid larvae (n=20) from 20 human patients who were undergone gastrointestinal endoscopy at a health check-up center in Korea, and molecular identification was performed on the larvae using PCR-RFLP analysis and gene sequencing of rDNA ITS regions and mtDNA cox2. In addition, anisakid larvae (n=53) collected from the sea eel (Astroconger myriaster) were also examined for comparison with those extracted from humans. The results showed that all human samples (100%) were identified as A. pegreffii, whereas 90.7% of the samples from the sea eel were A. pegreffii with the remaining 9.3% being Hysterothylacium aduncum. Our study confirmed that A. pegreffii is the predominant species causing human anisakiasis in Korea, and this seems to be due to the predominance of this larval type in the fish (sea eels) popularly consumed by the Korean people. The possibility of human infection with H. aduncum in Korea is also suggested.

• Parasites, Hosts and Diseases
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• 제52권4호
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• pp.383-389
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• 2014

Anisakiasis, a human infection of Anisakis L3 larvae, is one of the common foodborne parasitic diseases in Korea. Studies on the identification of anisakid larvae have been performed in the country, but most of them have been focused on morphological identification of the larvae. In this study, we analyzed the molecular characteristics of 174 Anisakis type I larvae collected from 10 species of fish caught in 3 different sea areas in Korea. PCR-RFLP and sequence analyses of rDNA ITS and mtDNA cox1 revealed that the larvae showed interesting distribution patterns depending on fish species and geographical locations. Anisakis pegreffii was predominant in fish from the Yellow Sea and the South Sea. Meanwhile, both A. pegreffii and A. simplex sensu stricto (A. simplex s.str.) larvae were identified in fish from the East Sea, depending on fish species infected. These results suggested that A. pegreffii was primarily distributed in a diverse species of fish in 3 sea areas around Korea, but A. simplex s.str. was dominantly identified in Oncorhynchus spp. in the East Sea.

• Fisheries and Aquatic Sciences
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• 제19권5호
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• pp.26.1-26.8
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• 2016

Background: The nematode species belonging to genus Anisakis occur at their third larval stage in numerous marine teleost fish species worldwide and known to cause accidental human infection through the ingestion of raw or undercooked fish or squids. They may also draw the attention of consumers because of the visual impact of both alive and dead worms. Therefore, the information on their geographical distribution and clear species identification is important for epidemiological survey and further prevention of human infection. Results: For identification of anisakid nematodes species isolated from largehead hairtail (Trichiurus japonicus), polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of internal transcribed spacers of ribosomal DNA were conducted. Mitochondrial cytochrome c oxidase subunit 2 gene was also sequenced, and phylogenetic analysis was conducted. From the largehead hairtail (n = 9), 1259 nematodes were isolated in total. Most of the nematodes were found encapsulated throughout the viscera (56.2 %, 708/1259) or moving freely in the body cavity (41.5 %, 523/1259), and only 0.3 % (4/1259) was found in the muscles. By PCR-RFLP, three different nematode species were identified. Anisakis pegreffii was the most dominantly found (98.7 %, 1243/1259) from the largehead hairtail, occupying 98.7 % (699/708) of the nematodes in the mesenteries and 98.1 % (513/523) in the body cavity. Hybrid genotype (Anisakis simplex ${\times}$ A. pegreffii) occupied 0.5 %, and Hysterothylacium sp. occupied 0.2 % of the nematodes isolated in this study. Conclusions: The largehead hairtail may not significantly contribute accidental human infection of anisakid nematode third stage larvae because most of the nematodes were found from the viscera or body cavity, which are not consumed raw. But, a high prevalence of anisakid nematode larvae in the largehead hairtail is still in concern because they may raise food safety problems to consumers. Immediate evisceration or freezing of fish after catch will be necessary before consumption.

• 한국어병학회지
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• 제25권3호
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• pp.189-197
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• 2012

2010~2012년까지 남해안 연안에서 서식하는 총 9목 30과 50종 243마리의 자연산 해산어류를 대상으로 아니사키스과 선충 유충의 감염현황을 조사하였다. 아니사키스과 선충 유충의 감염률은 10.7% (26/243마리) 로 나타났으며, 2010년 여수 시료에서 7.4% (7/95마리), 2011년 여수와 제주 시료에서 각각 22.7% (5/22마리) 와 8.2% (5/61마리), 2012년 완도 시료에서 40.9% (9/22마리) 로 나타났다. 2011년 통영 및 완도 시료에서는 아니사키스과 선충 유충이 발견되지 않았다. 총 10종의 어류 (성대, 쏨뱅이, 넙치, 볼락, 삼치, 흰꼬리볼락, 점농어, 황놀래기, 독가시치 및 노래미) 26마리에서 분리된 89마리의 아니사키스과 선충 유충을 ITS gene을 타겟으로 한 PCR-RFLP 및 염기서열 분석을 통해 동정한 결과, 6종의 아니사키스충이 동정되었고 성대, 쏨뱅이, 넙치 및 볼락에서 상대적으로 높은 감염율을 보였다: Anisakis pegreffii (53.9%, 48/89 아니사키스 유충), Hysterothylacium aduncum (38.2%, 34/89), H. fabri (3.4%, 3/89), hybird (A. simplex X A. pegreffii) (2.4%, 2/89), A. simplex (1.1%, 1/89), Raphidascaris lophii (1.1%, 1/89). 유충의 감염은 혼합감염 (19.2%, 5/26마리) 보다는 단독감염 (80.8%, 21/26마리) 이 높게 나타났다.

• Parasites, Hosts and Diseases
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• 제57권6호
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• pp.699-704
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• 2019

Anisakiasis (anisakidosis) refers to a foodborne zoonosis caused by ingesting raw or undercooked marine fish or cephalopods infected with anisakid larvae. The present study was performed to investigate the prevalence of anisakid larvae in anchovies (Engraulis japonica) purchased from 2 local markets in Gyeongsangnam-do, the Republic of Korea (=Korea), during 2018-2019. Anchovies were transported to our laboratory and examined by pepsin-HCl artificial digestion technique followed by microscopic observations and molecular analyses. The overall prevalence of anisakid larvae was 19.5% (39/200), from which a total of 51 larvae (av. 1.3 larvae/infected anchovy) were recovered. Sequencing of the larvae targeting the ITS region, including ITS1, 5.8S rRNA, and ITS2 genes confirmed the species of larvae as Anisakis pegreffii (54.9%; 28/51), Hysterothylacium sinense (23.5%; 12/51), and Hysterothylacium aduncum (21.5%; 11/51). The results suggested that anchovies could be a potential source of human anisakiasis in Korea.

• Parasites, Hosts and Diseases
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• 제58권1호
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• pp.15-25
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• 2020

The third stage larvae (L3) of Anisakis typica were detected in 2 species of threadfin bream, Nemipterus hexodon and N. japonicus, from the Gulf of Thailand, and were morphologically and molecularly characterized. Total 100 threadfin breams, 50 Nemipterus hexodon and 50 N. japonicus, were examined with naked eyes after the opening of abdominal cavity with scissors. Almost all infected larvae remained alive and active even the fish were transported for 1-2 days. Anisakid larvae were exclusively distributed in the body cavity and rarely in the liver. The prevalence of A. typica L3 were 68.0% and 60.0% in N. hexodon and N. japonicus and their infection intensities were 3.5 and 4.2 per fish infected each. Morphological and morphometric analysis were performed by viewing specimens under both a light microscope and a scanning electron microscope. Interestingly, the protruded mucron of Anisakis typica under SEM showed a distinct cylindrical shape that differed from the cone shape of A. simplex. The protruded mucron could be used to identify A. typica L3 larvae in the future. A comparison of the ITS1-5.8S-ITS2 rDNA nucleotide sequences of these species revealed high blast scores with A. typica. Conclusively, it was confirmed that A. typica L3 are prevalent in threadfin breams from the Gulf of Thailand, and their morphological and molecular characters are something different from those of other anisakid larvae, including A. simplex and A. pegreffii.

• Applied Microscopy
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• 제41권3호
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• pp.205-213
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• 2011

조피볼락, Sebastes schlegeli 소화관 주변에는 다수의 아니사키 스가 기생하고 있었으며, 몇몇 개체들은 간의 표면에 존재하고 있었다. 유전학적 결과 조피볼락에 기생한 아니사키스는 3종으로 분석되었으며, Hysterothylacium sp.가 가장 많이 분포하고 있었다. Hysterothylacium sp.는 Anisakis simplex보다 짧고 가늘었다. 두 종 모두 전방부분에는 lip, 입, 신경환, 분비공, 분비관이 관찰되었다. 두 종 모두 중간부분에는 식도, 위, 장이 존재하고 있었 지만, intestinal ceacum과 ventricular appendage는 Hysterothylacium sp.에서만 관찰되었다. Hysterothylacium sp.의 후방부분 끝에는 conical nodulose apex이 관찰되었지만, A. simplex의 후방부분 끝에는 spine이 관찰되었다. SEM으로 관찰한 결과 Hysterothylacium sp.의 입 주위에는 3개의 lip이 존재하고 있었는데, dorsal lip은 넓이 65 ${\mu}m$ 내외였고, 한쌍의 lateroventral lip은 넓이 60 ${\mu}m$ 내 외였다. Lip의 표면에는 넓이 8 ${\mu}m$ 내외의 유두돌기 한쌍이 존재하고 있었다. Hysterothylacium sp.와 A. simplex의 충체 넓이는 각각 480 ${\mu}m$와 900 ${\mu}m$ 내외로 측정되었다. Hysterothylacium sp. 의 lateral alae는 높이 7 ${\mu}m$ 내외였으며, cornical nodulose apex에 존재하는 돌기는 지름 약 3.3 ${\mu}m$였다. A. simplex의 후방부분 끝 에는 높이 약 20 ${\mu}m$의 spine이 관찰되었다. 조피볼락 장의 내강 에 존재하는 아니사키스들은 대부분 Hysterothylacium sp.였고, 점막주름 근처 혹은 점막하층에 기생하고 있었다. 아니사키스의 기생으로 인하여 숙주의 점막상피에는 다수의 점액세포가 증가하였다.