Quantitative detection of parasitic ciliate Cryptocaryon irritans in seawater with an optimized sample processing method
The protozoan Cryptocaryon irritans is one of the most important ectoparasites of marine fish, causing ‘white spot disease’ and mass mortality in aquaculture. To accurately predict disease outbreaks and develop prevention strategies, improved detection methods are required that are sensitive, conven...
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| Veröffentlicht in: | Journal of fish diseases 2022-05, Vol.45 (5), p.623-630 |
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| container_title | Journal of fish diseases |
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| creator | Tang, Jia‐Jia Zhong, Zhi‐Hong Li, Zhi‐Cheng Guo, Qing‐Kai Li, Shi‐Yu Guo, Yi‐Xuan Jiang, Biao Li, An‐Xing |
| description | The protozoan Cryptocaryon irritans is one of the most important ectoparasites of marine fish, causing ‘white spot disease’ and mass mortality in aquaculture. To accurately predict disease outbreaks and develop prevention strategies, improved detection methods are required that are sensitive, convenient and rapid. In this study, a pair of specific primers based on the C. irritans 18S rRNA gene was developed and used in a real‐time PCR (qPCR) assay. This assay was able to detect five theronts in 1 L of natural seawater. Furthermore, a linear model was established to analyse the log of Ct value and parasite abundance in seawater (y = –2.9623x + 24.2930), and the coefficient of determination (R2) value was 0.979. A lysis buffer was optimized for theront DNA extraction and used for storage sample. This method was superior to the commercial water DNA kit, and there was no significant degradation of DNA at room temperature for 24–96 hr. A dilution method was developed to manage qPCR inhibitors and used to investigate natural seawater samples in a net cage farm with diseased fish, and the findings were consistent with the actual situation. This study provides a valuable tool for assisting in the early monitoring and control of cryptocaryoniasis in aquaculture. |
| doi_str_mv | 10.1111/jfd.13587 |
| format | Article |
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To accurately predict disease outbreaks and develop prevention strategies, improved detection methods are required that are sensitive, convenient and rapid. In this study, a pair of specific primers based on the C. irritans 18S rRNA gene was developed and used in a real‐time PCR (qPCR) assay. This assay was able to detect five theronts in 1 L of natural seawater. Furthermore, a linear model was established to analyse the log of Ct value and parasite abundance in seawater (y = –2.9623x + 24.2930), and the coefficient of determination (R2) value was 0.979. A lysis buffer was optimized for theront DNA extraction and used for storage sample. This method was superior to the commercial water DNA kit, and there was no significant degradation of DNA at room temperature for 24–96 hr. A dilution method was developed to manage qPCR inhibitors and used to investigate natural seawater samples in a net cage farm with diseased fish, and the findings were consistent with the actual situation. This study provides a valuable tool for assisting in the early monitoring and control of cryptocaryoniasis in aquaculture.</description><identifier>ISSN: 0140-7775</identifier><identifier>EISSN: 1365-2761</identifier><identifier>DOI: 10.1111/jfd.13587</identifier><identifier>PMID: 35176179</identifier><language>eng</language><publisher>England: Blackwell Publishing Ltd</publisher><subject>Animals ; Aquaculture ; Chemical analysis ; Ciliophora ; Ciliophora Infections - diagnosis ; Ciliophora Infections - parasitology ; Ciliophora Infections - veterinary ; Cryptocaryon irritans ; Deoxyribonucleic acid ; Detection ; Dilution ; Disease outbreaks ; DNA ; Ectoparasites ; Fish ; Fish diseases ; Fish Diseases - parasitology ; Fish farms ; Husbandry diseases ; Lysis ; lysis buffer ; Marine fish ; Marine fishes ; Nucleotide sequence ; Parasites ; PCR ; Perciformes - parasitology ; Primers ; real‐time PCR ; Room temperature ; rRNA 18S ; sample storage ; Seawater ; Specimen Handling ; Storage ; Water analysis ; White spot disease</subject><ispartof>Journal of fish diseases, 2022-05, Vol.45 (5), p.623-630</ispartof><rights>2022 John Wiley & Sons Ltd</rights><rights>2022 John Wiley & Sons Ltd.</rights><rights>Copyright © 2022 John Wiley & Sons Ltd</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3537-333f13be17f62b72f06488b5663bb738ca681b6f737a59cde24cf680174cdc2b3</citedby><cites>FETCH-LOGICAL-c3537-333f13be17f62b72f06488b5663bb738ca681b6f737a59cde24cf680174cdc2b3</cites><orcidid>0000-0002-3851-7115</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fjfd.13587$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fjfd.13587$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27903,27904,45553,45554</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/35176179$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Tang, Jia‐Jia</creatorcontrib><creatorcontrib>Zhong, Zhi‐Hong</creatorcontrib><creatorcontrib>Li, Zhi‐Cheng</creatorcontrib><creatorcontrib>Guo, Qing‐Kai</creatorcontrib><creatorcontrib>Li, Shi‐Yu</creatorcontrib><creatorcontrib>Guo, Yi‐Xuan</creatorcontrib><creatorcontrib>Jiang, Biao</creatorcontrib><creatorcontrib>Li, An‐Xing</creatorcontrib><title>Quantitative detection of parasitic ciliate Cryptocaryon irritans in seawater with an optimized sample processing method</title><title>Journal of fish diseases</title><addtitle>J Fish Dis</addtitle><description>The protozoan Cryptocaryon irritans is one of the most important ectoparasites of marine fish, causing ‘white spot disease’ and mass mortality in aquaculture. To accurately predict disease outbreaks and develop prevention strategies, improved detection methods are required that are sensitive, convenient and rapid. In this study, a pair of specific primers based on the C. irritans 18S rRNA gene was developed and used in a real‐time PCR (qPCR) assay. This assay was able to detect five theronts in 1 L of natural seawater. Furthermore, a linear model was established to analyse the log of Ct value and parasite abundance in seawater (y = –2.9623x + 24.2930), and the coefficient of determination (R2) value was 0.979. A lysis buffer was optimized for theront DNA extraction and used for storage sample. This method was superior to the commercial water DNA kit, and there was no significant degradation of DNA at room temperature for 24–96 hr. A dilution method was developed to manage qPCR inhibitors and used to investigate natural seawater samples in a net cage farm with diseased fish, and the findings were consistent with the actual situation. This study provides a valuable tool for assisting in the early monitoring and control of cryptocaryoniasis in aquaculture.</description><subject>Animals</subject><subject>Aquaculture</subject><subject>Chemical analysis</subject><subject>Ciliophora</subject><subject>Ciliophora Infections - diagnosis</subject><subject>Ciliophora Infections - parasitology</subject><subject>Ciliophora Infections - veterinary</subject><subject>Cryptocaryon irritans</subject><subject>Deoxyribonucleic acid</subject><subject>Detection</subject><subject>Dilution</subject><subject>Disease outbreaks</subject><subject>DNA</subject><subject>Ectoparasites</subject><subject>Fish</subject><subject>Fish diseases</subject><subject>Fish Diseases - parasitology</subject><subject>Fish farms</subject><subject>Husbandry diseases</subject><subject>Lysis</subject><subject>lysis buffer</subject><subject>Marine fish</subject><subject>Marine fishes</subject><subject>Nucleotide sequence</subject><subject>Parasites</subject><subject>PCR</subject><subject>Perciformes - parasitology</subject><subject>Primers</subject><subject>real‐time PCR</subject><subject>Room temperature</subject><subject>rRNA 18S</subject><subject>sample storage</subject><subject>Seawater</subject><subject>Specimen Handling</subject><subject>Storage</subject><subject>Water analysis</subject><subject>White spot disease</subject><issn>0140-7775</issn><issn>1365-2761</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp10U1P4zAQBmALsaLl48AfQJa4wCFgx4mdHlGhsCukFRKcI8cZU1f5wnYo3V_PQGEPK60vc3nm1XiGkGPOLji-y5WtL7jIC7VDplzIPEmV5LtkynjGEqVUPiH7IawY4yrnco9MRM5RqNmUvD2Muosu6uhegdYQwUTXd7S3dNBeBxedocY1Tkegc78ZYm-036Bw3mNbF6jraAC9RuDp2sUl1dg-RNe6P1DToNuhATr43kAIrnumLcRlXx-SH1Y3AY6-6gF5Wtw8zu-S-9-3P-dX94kRuVCJEMJyUQFXVqaVSi2TWVFUuZSiqpQojJYFr6RVQul8ZmpIM2NlgT_NTG3SShyQs20uTvAyQohl64KBptEd9GMoUynYTPC0YEhP_6GrfvQdTocqU0zOMBXV-VYZ34fgwZaDdy3upOSs_DhHiecoP8-B9uQrcaxaqP_K7_0juNyCtWtg8_-k8tfiehv5Dns7lb0</recordid><startdate>202205</startdate><enddate>202205</enddate><creator>Tang, Jia‐Jia</creator><creator>Zhong, Zhi‐Hong</creator><creator>Li, Zhi‐Cheng</creator><creator>Guo, Qing‐Kai</creator><creator>Li, Shi‐Yu</creator><creator>Guo, Yi‐Xuan</creator><creator>Jiang, Biao</creator><creator>Li, An‐Xing</creator><general>Blackwell Publishing Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7TN</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>F1W</scope><scope>FR3</scope><scope>H94</scope><scope>H95</scope><scope>H98</scope><scope>H99</scope><scope>L.F</scope><scope>L.G</scope><scope>M7N</scope><scope>P64</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-3851-7115</orcidid></search><sort><creationdate>202205</creationdate><title>Quantitative detection of parasitic ciliate Cryptocaryon irritans in seawater with an optimized sample processing method</title><author>Tang, Jia‐Jia ; 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To accurately predict disease outbreaks and develop prevention strategies, improved detection methods are required that are sensitive, convenient and rapid. In this study, a pair of specific primers based on the C. irritans 18S rRNA gene was developed and used in a real‐time PCR (qPCR) assay. This assay was able to detect five theronts in 1 L of natural seawater. Furthermore, a linear model was established to analyse the log of Ct value and parasite abundance in seawater (y = –2.9623x + 24.2930), and the coefficient of determination (R2) value was 0.979. A lysis buffer was optimized for theront DNA extraction and used for storage sample. This method was superior to the commercial water DNA kit, and there was no significant degradation of DNA at room temperature for 24–96 hr. A dilution method was developed to manage qPCR inhibitors and used to investigate natural seawater samples in a net cage farm with diseased fish, and the findings were consistent with the actual situation. This study provides a valuable tool for assisting in the early monitoring and control of cryptocaryoniasis in aquaculture.</abstract><cop>England</cop><pub>Blackwell Publishing Ltd</pub><pmid>35176179</pmid><doi>10.1111/jfd.13587</doi><tpages>8</tpages><orcidid>https://orcid.org/0000-0002-3851-7115</orcidid></addata></record> |
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| ispartof | Journal of fish diseases, 2022-05, Vol.45 (5), p.623-630 |
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| subjects | Animals Aquaculture Chemical analysis Ciliophora Ciliophora Infections - diagnosis Ciliophora Infections - parasitology Ciliophora Infections - veterinary Cryptocaryon irritans Deoxyribonucleic acid Detection Dilution Disease outbreaks DNA Ectoparasites Fish Fish diseases Fish Diseases - parasitology Fish farms Husbandry diseases Lysis lysis buffer Marine fish Marine fishes Nucleotide sequence Parasites PCR Perciformes - parasitology Primers real‐time PCR Room temperature rRNA 18S sample storage Seawater Specimen Handling Storage Water analysis White spot disease |
| title | Quantitative detection of parasitic ciliate Cryptocaryon irritans in seawater with an optimized sample processing method |
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