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2024 Vol.4, Issue 1

Original Articles

Development of a Paralytic Shellfish Poisoning Detection Method Using Bioluminescent Bacterium, Aliivibrio fischeri

해양생물 중 마비성패독 검출을 위한 발광성 박테리아 Aliivibrio fischeri의 적용 연구

Youngjin Kim1
,
Kyoungah Lee1
,
Namhyun Kim1
,
Youngra Kim1
,
Kanghyun Lee1
,
Jinyoung Ma1
,
Jungsuk Lee1
,
Young-Seok Han1*
김 영진1
,
이 경아1
,
김 남현1
,
김 영라1
,
이 강현1
,
마 진영1
,
이 정석1
,
한 영석1*
1Institute of Environmental Protection and Safety, NEB Co., Bucheon 14523, Korea
1㈜엔이비, 환경바이오연구센터
*Corresponding Author
30 June 2024. pp. 1-9
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Abstract

Paralytic shellfish poisoning (PSP) occurs when seafood contaminated with toxin-producing dinoflagellates (e.g. Alexandrium species, Gymnodinium species and Pyrodinium bahamense) is consumed. Saxitoxin (STX). Paralytic shellfish toxin has more than 30 analogues, and its toxicity varies among the causative organisms. This study evaluated the differences in toxicity of STX and nine analogues (GTX1&4, GTX2&3, NEO, dcNEO, dcSTX, dcGTX2&3, C1&C2, GTX5, GTX6) using the Bioluminescence Inhibition Assay (BIA), and the toxicity test measured the inhibition rate of luminescence of bacterium, Aliivibrio fischeri. The toxicity index was calculated as EC50 (mg/L) values. The derived EC50 values were converted to Toxic Equivalency Factors (TEF) for the STX group, using STX (20.9 mg/L) as the reference. The BIA revealed that GTX6 exhibited the highest toxicity but the toxicity of GTX2&3 was the lowest. This result differed from the TEF values suggested by WHO (2016) and EFSA (2009), indicating that BIA may lack substrate specificity for STX group. Therefore, to propose an alternative screening method to the mouse bioassay, further research is needed to confirm the specificity of BIA for paralytic shellfish toxins.

Keywords
Aliivibrio fischeri
Bioluminescence Inhibition rate
Paralytic Shellfish Poisoning (PSP)
Saxitoxin (STX)
Toxic Equivalency Factor (TEF)

마비성패독에 의한 중독(Paralytic shellfish poisoning, PSP)은 와편모조류 등이 생성하는 독소에 노출된 수산물에 의해 발생한다. 대표적인 마비성패독 삭시톡신(Saxitoxin, STX)은 30개 이상의 이성질체가 있으며, 종류에 따라 독성에 차이가 있고 체내에서의 독성 성분의 변화를 일으키기도 한다. 본 연구는 발광박테리아 알리비브리오(Aliivibrio fischeri)를 활용해 STX군 독소들을 스크리닝하고 발광저해율에 따른 독성 차이를 확인하였다. 이 독성평가방법을 Bioluminescence inhibition assay (BIA)라고 하며, 발광박테리아의 발광저해율로 독성 유무를 확인한다. 실험은 STX 과 9종의 이성질체(STX, GTX1&4, GTX2&3, NEO, dcNEO, dcSTX, dcGTX2&3, C1&C2, GTX5, GTX6)의 표준독소로 BIA를 진행하였고, 독성지수로 EC50 (mg/L)값을 산출하였다. 산출된 EC50값은 STX (20.9 mg/L)을 기준으로 STX group의 독성을 독성등가계수(Toxic Equivalency Factor, TEF)로 치환하였다. BIA 결과로 GTX6가 가장 높은 독성을 GTX2&3이 가장 낮은 독성을 나타내었는데, 이는 WHO (2016), EFSA (2009)에서 제시한 TEF값과 상이한 것으로 나타났다. BIA를 이용한 독성평가 결과, 독소별 독성의 차이는 나타났지만 마비성패독에 대한 기질 특이성은 알려지지 않았다. BIA를 MBA 대체시험법으로 제시하기 위해서는 마비성패류독소에 작용에 대한 특이성을 확인할 수 있는 연구가 필요할 것으로 사료된다.

키워드
독성등가계수
마비성패독
발광박테리아
발광저해율
삭시톡신
References
1

Aballay-Gonzalez A, Ulloa V, Rivera A, Hernández V, Silva M, Caprile T, Delgado-Rivera L and Astuya A. 2016. Matrix effects on a cell-based assay used for the detection of paralytic shellfish toxins in bivalve shellfish samples. Food Additives & Contaminants: Part A 33, 869-875.

10.1080/19440049.2016.116674127002718
2

Anderson DM. 1997. Turning back the harmful red tide. Nature 388, 513-514.

10.1038/41415
3

Anderson DM, Alpermann TJ, Cembella AD, Collos Y, Masseret E and Montresor M. 2012. The globally distributed genus Alexandrium: multifaceted roles in marine ecosystems and impacts on human health. Harmful Algae 14, 10-35.

10.1016/j.hal.2011.10.01222308102PMC3269821
4

Blaghen M and Abakar Ah. A. 2019. Bioluminescence of Vibrio fischeri: A novel application for PSP quantification. Journal of Applied Biology and Biotechnology 7, 60-64.

10.7324/JABB.2019.70111
5

Bolelli L, Ferri EN and Girotti S. 2016. The management and exploitation of naturally light-emitting bacteria as a flexible analytical tool: A tutorial. Analytica Chimica Acta 934, 22-35.

10.1016/j.aca.2016.05.03827506340
6

David LS and Nicholson RA. 2004. Quantitation of paralytic shellfish toxins using mouse brain synaptoneurosomes. Chemosphere 55, 1315-1321.

10.1016/j.chemosphere.2003.10.05615081774
7

EFSA. 2009. Marine biotoxins in shellfish-Saxitoxin group. EFSA Journal 7, 1019.

10.2903/j.efsa.2009.1019
8

Egmond HP. 2004. Marine biotoxins 80. Food & Agriculture Org.

9

Fulladosa E, Murat J, Martínez M and Villaescusa I. 2005. Patterns of metals and arsenic poisoning in Vibrio fischeri bacteria. Chemosphere 60, 43-48.

10.1016/j.chemosphere.2004.12.02615910900
10

Girotti S, Bolelli L, Roda A, Gentilomi G and Musiani M. 2002. Improved detection of toxic chemicals using bioluminescent bacteria. Analytica Chimica Acta 471, 113-120.

10.1016/S0003-2670(02)00870-X
11

Jellett JF, Marks LJ, Stewart JE, Dorey ML, Watson-Wright W and Lawrence JF. 1992. Paralytic shellfish poison (saxitoxin family) bioassays: automated endpoint determination and standardization of the in vitro tissue culture bioassay, and comparison with the standard mouse bioassay. Toxicon 30, 1143-1156.

10.1016/0041-0101(92)90430-D1440621
12

Jellett JF, Roberts RL, Laycock MV, Quilliam MA and Barrett RE. 2002. Detection of paralytic shellfish poisoning (PSP) toxins in shellfish tissue using MIST AlertTM, a new rapid test, in parallel with the regulatory AOAC® mouse bioassay. Toxicon 40, 1407-1425.

10.1016/S0041-0101(02)00153-812368111
13

Jennings VL, Rayner-Brandes MH and Bird DJ. 2001. Assessing chemical toxicity with the bioluminescent photobacterium (Vibrio fischeri): a comparison of three commercial systems. Water Research 35, 3448-3456.

10.1016/S0043-1354(01)00067-711547867
14

Lee KT, Park GS and Kim PJ. 2008. Development of marine toxicity standard method for marine luminescent bacteria: Introduction of n-tox test. The Sea 13, 156-163.

15

Lee SY, Im JH, Woo SY, Choi HY, Park SB, Yoo CN and Chun HS. 2020. Saxitoxin and Its Analogues: Toxicity, Analytical Method, Occurrence and Safety Management. Journal of Food Hygiene and Safety 35, 521-534.

10.13103/JFHS.2020.35.6.521
16

Louzao MC, Vieytes MR, Cabado AG, Vieites Baptista de Sousa JM and Botana LM. 2003. A fluorimetric microplate assay for detection and quantitation of toxins causing paralytic shellfish poisoning. Chemical Research in Toxicology 16, 433-438.

10.1021/tx025574r12703959
17

Marugán J, Bru D, Pablos C and Catalá M. 2012. Comparative evaluation of acute toxicity by Vibrio fischeri and fern spore based bioassays in the follow-up of toxic chemicals degradation by photocatalysis. Journal of Hazardous Materials 213, 117-122.

10.1016/j.jhazmat.2012.01.07522341980
18

Nam KT and Oh SJ. 2021. Influence of water temperature and salinity on the production of paralytic shellfish poisoning by toxic dinoflagellate Alexandrium catenella (Group I). Journal of the Korean Society of Marine Environment & Safety 27, 119-126.

10.7837/kosomes.2021.27.1.119
19

NIFS. 2011. Improvement of Analysis Method for Paralytic Shellfish Poisoning Toxin.

20

NIFS. 2024. Shellfish Poisoning Report. Retrieved from https://www.nifs.go.kr/board/actionBoard0021List.do on May 7.

21

Quilliam MA, Thomson BA, Scott GJ and Siu KM. 1989. Ion‐spray mass spectrometry of marine neurotoxins. Rapid Communications in Mass Spectrometry 3, 145-150.

10.1002/rcm.12900305082520236
22

Schantz E. 1985. Purified shellfish poison for bioassay standardization. Conference on Shellfish Toxicology 1957.

23

Schantz EJ, Ghazarossian VE, Schnoes HK, Strong FM, Springer JP, Pezzanite JO, and Clardy J. 1975. Structure of saxitoxin. Journal of the American Chemical Society 97(5), 1238-1239.

10.1021/ja00838a0451133383
24

Scheerer S, Gomez F and Lloyd D. 2006. Bioluminescence of Vibrio fischeri in continuous culture: Optimal conditions for stability and intensity of photoemission. Journal of Microbiological Methods 67, 321-329.

10.1016/j.mimet.2006.04.01016750278
25

Shin HH, Li Z, Kim ES, Park JW and Lim WA. 2017. Which species, Alexandrium catenella (Group I) or A. pacificum (Group IV), is really responsible for past paralytic shellfish poisoning outbreaks in Jinhae-Masan Bay, Korea? Harmful Algae 68, 31-39.

10.1016/j.hal.2017.07.00628962988
26

Shin HH, Li Z, Kim HJ, Park BS, Lee J, Shin AY, Park TG, Lee KW, Han KH and Youn JY. 2021. Alexandrium catenella (Group I) and A. pacificum (Group IV) cyst germination, distribution, and toxicity in Jinhae-Masan Bay, Korea. Harmful Algae 110, 102-122.

10.1016/j.hal.2021.10212234887002
27

Shukla DM, Bajwa V, Gajic D and Saxena PK. 2020. Quorum sensing inhibition in Vibrio fischeri: an efficient system to assess antibacterial properties of medicinal plants and their volatile compounds. Integr. Food Nutr. Metab. 7, 1-9.

10.15761/IFNM.1000281
28

Song KC, Lee KJ, Yu HS, Mok JS, Kim JH, Lim KS and Lee MA. 2013. Paralytic Shellfish Poisoning (PSP) analysis using liquid chromatography-tandem mass spectrometry. Korean Journal of Fisheries and Aquatic Sciences 46, 154-159.

10.5657/KFAS.2013.0154
29

Turrell E, Lacaze J and Stobo L. 2007. Determination of paralytic shellfish poisoning (PSP) toxins in UK shellfish. Harmful Algae 6, 438-448.

10.1016/j.hal.2006.12.002
30

Viallon J, Chinain M and Darius HT. 2020. Revisiting the neuroblastoma cell-based assay (CBA-N2a) for the improved detection of marine toxins active on voltage gated sodium channels (VGSCs). Toxins 12, 281.

10.3390/toxins1205028132349302PMC7290318
31

WHO. 2016. Toxicity equivalence factors for marine biotoxins associated with bivalve molluscs.

Information
  • Publisher :The Korean Society of Phycology
  • Publisher(Ko) :한국조류학회
  • Journal Title :Aquatic Nature
  • Journal Title(Ko) :수생생물
  • Volume : 4
  • No :1
  • Pages :1-9
  • Received Date : 2024-06-05
  • Revised Date : 2024-06-17
  • Accepted Date : 2024-06-17
  • DOI :https://doi.org/10.23135/an.2024.4.1.1
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