A technique for the preservation of Cryptocaryon irritans at low temperatures

Dan, Xue-Ming; Lin, Xiaotan; YuChen, Yan; Teng, N.; Tan, Zhijun; Li, A.X.

doi:10.1016/j.aquaculture.2009.10.001

Cited by 18 publications

(15 citation statements)

References 11 publications

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“…Although the recovery rate is one of the important issues to be resolved, it appears to be acceptable in this assay: it is important to estimate the order of magnitude, not the precise number of C. irritans cells for this disease. Dan et al [17,18] have shown that the lethal dose of theronts is more than 12,000 theronts per fish for the host pompano Trachinotus ovatus. Thus, the lethal dose, which is much higher than our detection limit (20 theronts), allows us to ignore the recovery rate.…”

Section: Discussionmentioning

confidence: 98%

Quantitative PCR assay for the detection of the parasitic ciliate Cryptocaryon irritans

2011

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We developed a quantitative PCR assay for detecting the parasitic ciliate Cryptocaryon irritans, which causes ''white spot disease'' in marine fishes, from the natural environment. A specific primer set for C. irritans was designed and its high specificity was confirmed in silico: almost all of the sequences deposited in the GenBank nucleotide database were covered, 22/23 for the forward primer and 7/7 for the reverse primer. We estimated that there were 3,415.9 rRNA gene copies per genome of C. irritans. In artificial mixture experiments to validate whether the qPCR assay is applicable to natural samples, the estimated copy numbers showed significantly positive correlations with the number of theronts added (p \ 0.001). When we applied this qPCR assay to natural samples collected bimonthly from surface and bottom seawaters at an aquaculture site (water depth, 10 m) from

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Section: Discussionmentioning

confidence: 98%

Quantitative PCR assay for the detection of the parasitic ciliate Cryptocaryon irritans

2011

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“…; Dan et al . ). The use of recombinant proteins allows antigens with uniform specificity obtained in large quantities at lower costs and less manpower (Ndifuna et al .…”

Section: Discussionmentioning

confidence: 97%

“…Even though the recombinant proteins produced were insoluble and purified under denaturing conditions, these proteins were suitable to be used in a cryptocaryonosis screen, specifically the CN56, CN57, CN72 and CN246 recombinant proteins that were identified as promising molecular markers. The utility of these proteins supersedes the use of antigens extracted from C. irritans as obtaining homogenous parasite stocks is difficult due to antigenic variation and strain heterogeneity, inability to propagate C. irritans by in vitro methods and difficulties in obtaining stable parasite stocks through in vivo propagation (Dickerson & Clark 1996;Yoshinaga et al 2007;Dan et al 2009). The use of recombinant proteins allows antigens with uniform specificity obtained in large quantities at lower costs and less manpower (Ndifuna et al 1998;Liljeqvist & Stahl 1999).…”

Section: Figurementioning

confidence: 99%

Cryptocaryon irritans recombinant proteins as potential antigens for sero‐surveillance of cryptocaryonosis

Lokanathan

Mohd-Adnan

et al. 2016

Journal of Fish Diseases

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Cryptocaryonosis is a major problem for mariculture, and the absence of suitable sero-surveillance tools for the detection of cryptocaryonosis makes it difficult to screen Cryptocaryon irritans-infected fish, particularly asymptomatic fish. In this study, we proposed a serum-based assay using selected C. irritans proteins to screen infected and asymptomatic fish. Eight highly expressed genes were chosen from an earlier study on C. irritans expressed sequence tags and ciliate glutamine codons were converted to universal glutamine codons. The chemically synthesized C. irritans genes were then expressed in an Escherichia coli expression host under optimized conditions. Five C. irritans proteins were successfully expressed in E. coli and purified by affinity chromatography. These proteins were used as antigens in an enzyme-linked immunosorbent assay (ELISA) to screen sera from experimentally immunized fish and naturally infected fish. Sera from both categories of fish reacted equally well with the expressed C. irritans recombinant proteins as well as with sonicated theronts. This study demonstrated the utility of producing ciliate recombinant proteins in a heterologous expression host. An ELISA was successfully developed to diagnose infected and asymptomatic fish using the recombinant proteins as antigens.

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“…Usaha memisahkan dari inangnya dan mengembangbiakannya secara in vitro telah dilakukan oleh beberapa peneliti namun masih belum memberikan hasil yang memuaskan. Oleh karena itu untuk saat ini pemeliharaan secara in vitro lebih ditujukan untuk memperpanjang masa simpan parasit (Dan et al, 2009). Meskipun tingkat patogenisitas I. multifiliis juga ditentukan oleh faktor lain seperti pH dan kesadahan air budidaya saat infeksi terjadi (Garcia et al, 2011), namun perlakuan saat penyimpanan parasit diduga juga memiliki pengaruh terhadap patogensisitas parasit.…”

Section: Abstrakunclassified

“…Abnormalitas parasit yang teramati antara lain mobilitas sel yang rendah, dan ukuran sel tomite yang tidak seragam. Penelitian sebelumnya yang telah dilakukan oleh Dan et al, (2009) pada parasit Cryptocaryon irritans juga menemukan gejala yang sama.…”

Section: Hasil Dan Pembahasanunclassified

Infektivitas parasit Ichtyophthirius multifiliis yang disimpan pada suhu rendah

Rahman

Sukenda

Nuryati³

et al. 2016

Jurnal Akuakultur Indonesia

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ABSTRACT The aim of this study was to evaluate infectivity of Ichtyophthirius multifiliis which caused white spot disease maintained at low temperature without its host. Briefly, the trophont stage of parasites were subjected at control (28 °C) and lower temperature (9 °C) for 14 consecutive days of observation. The rate of survival, and excystment of descendants were examined descriptively at the last day of observation. Here, the infectivity of parasite then performed by means infecting the model fish Poecilia sphenops (black moly) with escaping theronts. The results revealed that the survival rate and excystment rate of parasite were decreased as maintaining period increased. The final rate of survival, and excystment of parasite were 35% and 33,3% respectively. Additionally, the descendants came out with high abnormality which recognized by weak mobility and lower infectivity (50%) compared to the control (80%). Then, it is concluded that, maintaining I. multifiliis at low temperature without its host for 14 consecutive days will decreased the infectivity. Keywords: white spot, obligat parasite, excystment, infectivity ABSTRAK Penelitian ini bertujuan untuk mengevaluasi infektivitas parasit Ichtyophthirius multifiliis penyebab penyakit bintik putih (white spot) yang dipelihara tanpa inang pada suhu rendah. Parasit dengan stadia trophont dipelihara pada suhu ruang (28 °C) dan suhu rendah (9 °C) selama 14 hari. Selama masa pemeliharaan tersebut tingkat kelulusan hidup, dan tingkat eksismen parasit diukur dan dibandingkan secara deskriptif. Hari terakhir pemeliharaan dilakukan uji tantang pada ikan black moly Poecilia sphenops untuk menilai infektivitas parasit. Hasil penelitian memperlihatkan bahwa tingkat kelulusan hidup dan eksismen parasit semakin menurun dengan bertambahnya masa pemeliharaan. Akhir pengamatan kelangsungan hidup, dan nilai eksismen tersebut berturut-turut adalah 35% dan 33%. Parasit yang disimpan pada suhu rendah selama 14 hari memperlihatkan infektivitas yang lebih rendah (50%) dibandingkan dengan perlakuan kontrol (80%). Kesimpulannya, penyimpanan parasit I. multifiliis pada suhu rendah selama 14 hari dapat menurunkan infektivitas parasit pada inang. Kata kunci: bintik putih, parasit obligat, eksismen, infektivitas

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A technique for the preservation of Cryptocaryon irritans at low temperatures

Cited by 18 publications

References 11 publications

Quantitative PCR assay for the detection of the parasitic ciliate Cryptocaryon irritans

Quantitative PCR assay for the detection of the parasitic ciliate Cryptocaryon irritans

Cryptocaryon irritans recombinant proteins as potential antigens for sero‐surveillance of cryptocaryonosis

Infektivitas parasit Ichtyophthirius multifiliis yang disimpan pada suhu rendah

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