• Journal of Aquaculture
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• v.19 no.3
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• pp.197-204
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• 2006

The pathogenicity and infection route of the Scuticociliate, Philasterdies dicentrarchi, were investigated with the 3 and 5 cm-group of juvenile flounders, Paralichthys olivaceus. The infection rates of 3 cm-group were 40% four days post infection (D.P.I.) and increased to be 90.1% 24 D.P.I., whereas those of 5 cm-group were 20% 8 D.P.I., 42% 16 D.P.I., and 81% 24 D.P.I. The results showed there were several infection routes to internal organs Olive flounder. The first route was started with the infection at the soft part of caudal fin and later reached at fin ray and muscle tissue; the second one was started from lips and mouth tissue of upper jaw and later the pathogen could be observed at either muscle tissue or eyeball and brain; the third one was begun at caudal fin and later the pathogen reached at brain tissu'e through spiral cord; the fourth one was started with the infections at abdominal cavity and anus. P. dicentrarchi infected to brain tissue was first observed 14 D.P.I in 3 cm-group and 20 D.P.I. in 5 cm-group of the juvenile flounder. This indicated that the brain infection of P. dicentrarchi might occur faster in small-sized flounder than large-sized one.

• Journal of fish pathology
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• v.22 no.1
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• pp.45-52
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• 2009

Zebrafish was firstly applied to an experimental model for scuticociliatosis caused by Philasterides dicentrarchi, a facultative parasitic ciliate in cultured marine fish. The susceptibility of zebrafish to infection of P. dicentrarchi was assessed by intraperitoneal injection of the ciliates, which produced typical symptoms of scuticociliatosis and significant mortality. The potential use of zebrafish as a model to evaluate the vaccine efficacy against scuticociliatosis was analyzed by immunization of zebrafish with the ciliates lysate. Furthermore, the effect of different adjuvants, such as Quillaja saponin (QS), Montanide, and Freund’s incomplete adjuvant (FIA) on the protective efficacy of the vaccine was investigated. Groups of zebrafish injected with QS or Montanide alone showed higher survival of fish against challenge test compared to control fish. The results suggest that adjuvant-mediated enhancement of innate immune responses play important roles in protection of fish against scuticociliatosis. The considerably high survival in the fish immunized with the antigen alone indicates that the ciliate lysate itself is highly immunogenic to zebrafish, which can elicit protective immune responses. The protective potential of the antigen, ciliate lysate, was enforced through combined administration with adjuvants including QS, Montinide and FIA. No or low mortalities in the groups of fish immunized with the antigen plus adjuvants suggests that the adaptive immune responses of zebrafish might be accelerated by the adjuvants or the protective potential of the antigen and adjuvants might synergistically interact. In spite of several shortcomings such as difficulties in sampling of serum and leucocytes enough to routine immunological analyses, zebrafsih might be the most convenient experimental animal for scuticociliatosis.

• Journal of Aquaculture
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• v.20 no.2
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• pp.106-113
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• 2007

Biological characteristics of Philasteridies dicentrarchi (Ciliophora:Scuticociliate) isolated from the cultured olive flounder Paralichthys olivaceus was determined out by culture, growth conditions, and in vitro relief effect of chemical compounds. The scuticociliate has an active propagation ability by utilizing organic matters obtained from cell strain, bacteria, assorted feed, brain tissue and rotifer tissue. The ciliate achieved population growth activity under the conditions of $5{\sim}45\;ppt$ in salinity and pH 6-9. The ciliate had survived and propagated under the water temperature ranging $10{\sim}30^{\circ}C$, but active growth was observed in the temperature ranges of $10{\sim}25^{\circ}C$. Therapeutic trials were performed with formalin and hydrogen peroxide. The extermination time of the parasites with formalin was in 30 minutes both at 300 and 400 ppm, 60 minutes at 200 ppm, 90 minutes at 100 ppm, and 120 minutes at 50 ppm, respectively. In hydrogen peroxide treatments extermination time was 60 minutes in 300 ppm, 90 minutes in 200 ppm, and 150 minutes at 150 ppm and 100 ppm concentrations.

• Journal of fish pathology
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• v.17 no.2
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• pp.145-149
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• 2004

The definitive identification of ciliate species by morphological characteristics relies on time-consuming and laborious staining techniques. Therefore, in this study, we discriminated 3 scuticociliatosis causing species - Pseudocohnilembus persalinus, Uronema marinum and Philasterides dicentrarchi - in cultured olive flounder by multiplex PCR. The multiplex PCR based on the species-specific amplification of small subunit ribosomal RNA (SS rRNA) gene sequence enabled us to distinguish the 3 scuticociliate species in a simple and rapid manner, even in the sample containing the three species simultaneously. These data suggest that the multiplex PCR strategy would make it possible to avoid the cumbersome and time-consuming procedures of morphological analysis for the definitive identification of scuticociliates.

• Journal of fish pathology
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• v.19 no.2
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• pp.87-97
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• 2006

In previous study, scuticociliate P. dicentrachi was isolated from farmed flounder during the december 2004 to april 2005. Pathogenicity, mortality and infection symptom were studied using 3 cm and 5 cm groups of juvenile flounder. The ciliates were exponentially increased from bottom layer of the experimental tanks, which propagated within 2000 cells/ml (1.4 × 103 cell ㎖-1 ～ 2.5 × 103 cell ㎖-1) after 6 days of inoculation. The middle layer was maintained within 300 cells/ml. Both 3 cm and 5 cm groups were infected with ciliates, mainly 3 cm group showed high mortality during the experimental period. The death of 3 cm group was started from 5 days and showed 95.6% of mortality after 28 days of first inoculation. The control group showed 4.4% of mortality however, we could not observed any ciliates. The death of 5 cm group was started later than 3 cm group after 18 days of first inoculation. The total mortality was 71% during 28 days. No mortality and infection symptoms were observed in the control. We also studied SSU rRNA gene of ciliates which, re-isolated from infected flounder of experimental groups. When SSU rRNA in this study compared with previous data showed that the identified strain of both previous and present study was same.

• Journal of fish pathology
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• v.22 no.2
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• pp.97-105
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• 2009

Scuticociliates Miamiensis avidus (syn. Philasterides dicentrarchi) causes high mortality and bad growth in olive flounder Paralichthys olivaceus. Temperature is an important factor not only for growth of pathogens but also for host immune system in poikilothermal animal. In this study, temperature affecting ciliate growth and pathogenicity against olive flounder were examined. Doubling time for the ciliate growth was 61.82 hours at $5{^{\circ}C}$, 26.32 hours at $10{^{\circ}C}$, 21.14 hours at $15{^{\circ}C}$, 16.86 hours $20{^{\circ}C}$ and 16.21 hours at $25{^{\circ}C}$. Maximum ciliate numbers were similar at $10-20{^{\circ}C}$ at the range of $1.54-1.75{\times}10^{5}$/ ml. Duplicated intraperitoneal injections were conducted with the ciliates by the concentrations of $1{\times}10^{2}$, $1{\times}10^{6}$, $1{\times}10^{4}$and $1{\times}10^{5}$/ fish (average 8.34 cm, 4.33 g) then kept at $10{^{\circ}C}$, $15{^{\circ}C}$ and $20{^{\circ}C}$. Cumulative mortality was low at $10{^{\circ}C}$ and the mortality was increasing at higher water temperatures. In addition, cumulative mortality was higher at higher dose of infections. In conclusion, Scuticocilite M. avidus grew well at higher temperature (at $5{^{\circ}C}$, $10{^{\circ}C}$, $15{^{\circ}C}$ and $25{^{\circ}C}$) in vitro, and olive flounder mortality due to M. avidus was highly water temperature and dose dependent. The results of this study suggest that water temperature control may one of the essential factor to reduce mortality due to M. avidus infection.

• Journal of fish pathology
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• v.34 no.1
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• pp.47-53
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• 2021

Scuticociliates are one of the serious parasitic threats faced by the marine aquaculturists worldwide. To date, Uronema nigricans, Philasterides dicentrarchi, Miamiensis avidus, Uronema marinum, and Pseudocohnilembus persalinus have been reported as the important culprit species causing scuticociliatosis in fish. The present paper reports the finding of an additional scuticociliate isolate from the gill of diseased olive flounder Paralichthys olivaceus in Korea. Based on the morphological characteristics, a scuticociliate in this study was identified as Paranophrys marina. Phylogenetic analysis placed P. marina as a sister lineage to three species of Pseudocohnilembus and Mesanophrys carcini within the order Philasterida.