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Molecular identification and real-time quantitative PCR (qPCR) for rapid detection of Thelohanellus kitauei, a Myxozoan parasite causing intestinal giant cystic disease in the Israel carp.

Seo JS, Jeon EJ, Kim MS, Woo SH, Kim JD, Jung SH, Park MA, Jee BY, Kim JW, Kim YC, Lee EH - Korean J. Parasitol. (2012)

Bottom Line: Until now, studies concerning IGCD caused by T. kitauei in the Israel carp have been limited to morphological and histopathological examinations.In this study, we cloned the full-length 18S rRNA gene of T. kitauei isolated from diseased Israel carps, and carried out molecular identification by comparing the sequence with those of other myxosporeans.Our results demonstrated that both the conventional PCR and real-time quantitative PCR systems applied herein are effective for rapid detection of T. kitauei spores in fish tissues and environmental water.

View Article: PubMed Central - PubMed

Affiliation: Pathology Division, National Fisheries Research and Development Institute, Busan 619-705, Korea.

ABSTRACT
Intestinal giant-cystic disease (IGCD) of the Israel carp (Cyprinus carpio nudus) has been recognized as one of the most serious diseases afflicting inland farmed fish in the Republic of Korea, and Thelohanellus kitauei has been identified as the causative agent of the disease. Until now, studies concerning IGCD caused by T. kitauei in the Israel carp have been limited to morphological and histopathological examinations. However, these types of diagnostic examinations are relatively time-consuming, and the infection frequently cannot be detected in its early stages. In this study, we cloned the full-length 18S rRNA gene of T. kitauei isolated from diseased Israel carps, and carried out molecular identification by comparing the sequence with those of other myxosporeans. Moreover, conventional PCR and real-time quantitative PCR (qPCR) using oligonucleotide primers for the amplification of 18S rRNA gene fragment were established for further use as methods for rapid diagnosis of IGCD. Our results demonstrated that both the conventional PCR and real-time quantitative PCR systems applied herein are effective for rapid detection of T. kitauei spores in fish tissues and environmental water.

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Characterization of spores. (A) Various sizes of giant cysts (arrows) containing mature spores found in the intestine of an Israel carp infected with Thelohanellus kitauei. (B) Spores of T. kitauei isolated from the intestine of a diseased fish. Scale bar: 20 µm.
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Figure 1: Characterization of spores. (A) Various sizes of giant cysts (arrows) containing mature spores found in the intestine of an Israel carp infected with Thelohanellus kitauei. (B) Spores of T. kitauei isolated from the intestine of a diseased fish. Scale bar: 20 µm.

Mentions: The egg-shaped giant cysts (12-45 mm size) removed from the intestines of infected Israel carps were filled with numerous mature spores (Fig. 1A). The spores released from the cyst evidenced the typical specific features of T. kitauei, including an egg-shaped balloon-like sack. The size of spores, spore sacks, polar capsules, and polar filament were identified in accordance with the descriptions provided by Egusa and Nakajima [2] and Chun et al. [3]. We also observed spores in front (Fig. 1Bc), spores in side view (Fig. 1Bb), opened spore sacks, and divided spores within the sack (Fig. 1Ba, d) as previously described [2].


Molecular identification and real-time quantitative PCR (qPCR) for rapid detection of Thelohanellus kitauei, a Myxozoan parasite causing intestinal giant cystic disease in the Israel carp.

Seo JS, Jeon EJ, Kim MS, Woo SH, Kim JD, Jung SH, Park MA, Jee BY, Kim JW, Kim YC, Lee EH - Korean J. Parasitol. (2012)

Characterization of spores. (A) Various sizes of giant cysts (arrows) containing mature spores found in the intestine of an Israel carp infected with Thelohanellus kitauei. (B) Spores of T. kitauei isolated from the intestine of a diseased fish. Scale bar: 20 µm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3375447&req=5

Figure 1: Characterization of spores. (A) Various sizes of giant cysts (arrows) containing mature spores found in the intestine of an Israel carp infected with Thelohanellus kitauei. (B) Spores of T. kitauei isolated from the intestine of a diseased fish. Scale bar: 20 µm.
Mentions: The egg-shaped giant cysts (12-45 mm size) removed from the intestines of infected Israel carps were filled with numerous mature spores (Fig. 1A). The spores released from the cyst evidenced the typical specific features of T. kitauei, including an egg-shaped balloon-like sack. The size of spores, spore sacks, polar capsules, and polar filament were identified in accordance with the descriptions provided by Egusa and Nakajima [2] and Chun et al. [3]. We also observed spores in front (Fig. 1Bc), spores in side view (Fig. 1Bb), opened spore sacks, and divided spores within the sack (Fig. 1Ba, d) as previously described [2].

Bottom Line: Until now, studies concerning IGCD caused by T. kitauei in the Israel carp have been limited to morphological and histopathological examinations.In this study, we cloned the full-length 18S rRNA gene of T. kitauei isolated from diseased Israel carps, and carried out molecular identification by comparing the sequence with those of other myxosporeans.Our results demonstrated that both the conventional PCR and real-time quantitative PCR systems applied herein are effective for rapid detection of T. kitauei spores in fish tissues and environmental water.

View Article: PubMed Central - PubMed

Affiliation: Pathology Division, National Fisheries Research and Development Institute, Busan 619-705, Korea.

ABSTRACT
Intestinal giant-cystic disease (IGCD) of the Israel carp (Cyprinus carpio nudus) has been recognized as one of the most serious diseases afflicting inland farmed fish in the Republic of Korea, and Thelohanellus kitauei has been identified as the causative agent of the disease. Until now, studies concerning IGCD caused by T. kitauei in the Israel carp have been limited to morphological and histopathological examinations. However, these types of diagnostic examinations are relatively time-consuming, and the infection frequently cannot be detected in its early stages. In this study, we cloned the full-length 18S rRNA gene of T. kitauei isolated from diseased Israel carps, and carried out molecular identification by comparing the sequence with those of other myxosporeans. Moreover, conventional PCR and real-time quantitative PCR (qPCR) using oligonucleotide primers for the amplification of 18S rRNA gene fragment were established for further use as methods for rapid diagnosis of IGCD. Our results demonstrated that both the conventional PCR and real-time quantitative PCR systems applied herein are effective for rapid detection of T. kitauei spores in fish tissues and environmental water.

Show MeSH
Related in: MedlinePlus