• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.54 no.5
• /
• pp.652-659
• /
• 2021

Microorganism-derived compounds, such as peptidoglycan, lipoteichoic acid, and β-glucan were supplemented in the scuticociliate Miamiensis avidus (M. avidus) vaccine to verify the specify component contribution to the adjuvant effect. Vaccine was formulated with the inactivated M. avidus antigen (YS2, 4.44×10⁵ cells/fish) in combination with either peptidoglycan (10 ㎍ and 100 ㎍/fish), lipoteichoic acid (5 ㎍ and 50 ㎍/fish), or β-glucan (10 ㎍ and 100 ㎍/fish). Olive flounder injected with peptidoglycan supplemented vaccine (10 ㎍ and 100 ㎍/fish) exhibited significant protection, and the relative percent survival (RPS) was 55% and 65% at 4 weeks post vaccination (wpv), respectively, at the corresponding doses. The vaccine groups with added lipoteichoic acid (5 ㎍ and 50 ㎍/fish) exhibited RPS of 40% and 5%, respectively. Additionally, the group with added β-glucan (100 ㎍/fish) exhibited RPS of 35%, but no effect was observed in the group with added 10 ㎍/fish β-glucan. At 8 wpv, olive flounder injected with peptidoglycan and lipoteichoic acid supplemented vaccines exhibited protection with RPS range of 11/11% and 5/21%, respectively, at the respective doses. M. avidus vaccine containing 10 ㎍ and 100 ㎍/fish of β-glucan exhibited the RPS of 32% and 37%, respectively. Conclusively, peptidoglycan contributed in high protection of the M. avidus vaccine, and thus, it can be used as an effective adjuvant in the M. avidus vaccine.

• Fisheries and Aquatic Sciences
• /
• v.26 no.10
• /
• pp.605-616
• /
• 2023

Scuticociliatosis, caused by the parasitic pathogen Miamiensis avidus, poses a significant threat to olive flounder farms in South Korea. Infected fish suffer from severe systemic infections affecting various organs, with potential secondary bacterial diseases. This study investigated the emergence of different M. avidus serotypes in 20 olive flounder farms on Jeju island, South Korea, from 2015 to 2020. Additionally, we identified Tenacibaculum maritimum as a co-infecting bacteria. Based on serotyping and monitoring data, we developed a trivalent vaccine targeting two serotypes of M. avidus and one strain of T. maritimum. The efficacy of the vaccine was evaluated under laboratory conditions and demonstrated a relative percentage of survival (RPS) of 75%, 80%, and 93% for M. avidus serotype I, M. avidus serotype II and T. maritimum, respectively. Furthermore, successful field trials conducted on four different olive flounder farms resulted in significantly higher survival rates (52%-76% RPS) and weight gains in vaccinated fish. Overall, this study presents an effective vaccine against M. avidus and T. maritimum infections in farmed olive flounder, making a valuable contribution to sustainable aquaculture in South Korea.

• Journal of fish pathology
• /
• v.28 no.3
• /
• pp.171-174
• /
• 2015

Previously, we had reported that some Miamiensis avidus, a major pathogen of scuticociliatosis in cultured olive flounder, strongly agglutinated by flounder immune sera could escape from the agglutinated mass within a few hours. In the present study, we observed that M. avidus not only escaped from the agglutinated mass but also conducted division(s) before shedding its old covering. Furthermore, ciliates that survived the antibody-dependent complement killing (ADCK) assay were not killed even when re-exposed to a freshly prepared ADCK assay. This result suggests that the liberated ciliates from the ADCK assay might change not only their i-antigen types but also the epitopes of major surface antigens, which debilitate antibody-mediated complement killing ability.

• Journal of fish pathology
• /
• v.31 no.2
• /
• pp.109-113
• /
• 2018

The purpose of the present study was to determine whether the scuticocidal activity of olive flounder (Paralichthys olivaceus) head-kidney leucocytes can be enhanced by stimulation with polyinosine-polycytosine [poly (I:C)]. The growth of Miamiensis avidus was not affected by exposure to unstimulated or poly (I:C)-stimulated leucocytes alone, heat-inactivated immune serum alone, or unstimulated leucocytes plus heat-inactivated immune serum. However, leucocytes stimulated with poly (I:C) showed clearly high scuticocidal activity against M. avidus in the presence of heat-inactivated immune serum. Furthermore, numerous poly (I:C)-stimulated leucocytes occupied the surface of scuticociliates in the presence of the heat-inactivated immune serum, which led to lysis of scuticociliates. These results suggest that both of the stimulation of leukocytes and the immobilization of scuticociliates are necessary to kill scuticociliates by leukocytes.

• Journal of fish pathology
• /
• v.31 no.1
• /
• pp.15-21
• /
• 2018

Differences in in vivo virulence and in sensitivity to drugs among different isolates of Miamiensis avidus were analyzed. The isolate III showed the highest resistance against the scuticocidal activity of olive flounder (Paralichthys olivaceus) serum, and induced the highest mortalities of olive flounder fingerlings. The isolate II showed significantly higher serum resistance than the isolate I, but in vivo virulence of isolate II was not significantly different from that of isolate I. The secreted proteinases activity of isolate III was significantly higher than that of isolate I and II, and the activity was significantly reduced by the addition of E-64, a cysteine proteinases inhibitor. There were no differences among isolates in the sensitivity to doxycycline, however, there were significant differences in sensitivities to mebendazole and bithionol. These results suggest that the different characteristics of different M. avidus isolates should be taken into consideration for the development of control measures against scuticociliatosis.

• Journal of fish pathology
• /
• v.22 no.2
• /
• pp.97-105
• /
• 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
• /
• v.36 no.2
• /
• pp.311-321
• /
• 2023

The efficacy of the alginate microsphere (Alginate MS) oral vaccine against Miamiensis avidus in olive flounder (Paralichthys olivaceus) was confirmed through challenge infections by both immersion and injection routes. In trial 1, the formalin-inactivated M. avidus coated with alginate, designated as 'IMa+Alginate MS' group, and the IMa group were administered with vaccines mixed with feed, with a total antigen dose of 3.75 × 10⁶ cells/fish. When challenged with immersion infection at five weeks post vaccination, the relative percent survival (RPS) in the IMa+Alginate MS group was 50% (immersed in 50% seawater) and 37.5% (immersed in 100% seawater). The group that received only IMa showed a low survival rate. In trial 2, the antigen was fed mixed with feed at a total dose of 2.38 × 10⁶ cells/fish for 5 days. Two weeks after oral vaccination, fish were intraperitoneally injected for infection. The RPS in the IMa+Alginate MS group was 30.8%, while the IMa-only group showed no vaccine efficacy. At five weeks post vaccination, when subjected to challenge infection by immersion in 50% seawater, the IMa+Alginate MS group recorded a RPS of 42.9%, whereas the IMa group had a RPS of 14.3%. The results of this study indicate that coating M. avidus antigen with alginate can provide higher protection in olive flounder compared to administering the antigen alone.

• Journal of fish pathology
• /
• v.19 no.2
• /
• pp.173-181
• /
• 2006

Miamiensis avidus is a scuticociliate causing mortality in olive flounder Paralichthys olivaceus. To evaluate immune response of olive flounder against M. avidus, 2.6×106cells/fish of Formalin killed cell (FKC) was intraperitoneally (i.p.) injected, and 2.4 × 106cells/㎖ of sonicated FKC was immersion immunized to 14.9 cm (26.8g) fish. Fish were immunized 2 times with 2 weeks intervals. Antiserum from immunized fish caused agglutination and immobilization of the ciliate. In ELISA test, immunized group exhibited higher titers than control group. In addition, i.p. immunized fish showed higher phagocytic ratio, phahgocytic index and chemotaxic activity. To evaluate in vivo efficacy of FKC on fish, 1.43 × 105cells/fish was i.p.. injected, and 2.2 × 105 cells/㎖ of sonicated cell was immersed into 8.6cm (6.3g) fish. After 2 times immunization with 2 weeks intervals, fish were infected with 2.0 × 104 and 2.0 × 103 cells/ml of live ciliates by immersion. After 3 weeks, cumulative mortality was lower in the i.p. immunized group and mortality was delayed in immersion immunized group. In conclusion, specific immune response of oliver flounder against M. avidus was elevated after immunization and these immune response may prevent and/or delay for the M. avidus infection to olive flounder.

• Journal of fish pathology
• /
• v.26 no.2
• /
• pp.65-75
• /
• 2013

Population growth equation of scuticociliate Miamiensis avidus was obtained from the experimental results of in vitro culture condition to estimate the growth rate and carrying capacity from the growth equation. In addition, intraperitoneal infections into olive flounder Paralichthys olivaceus were carried out into 2 different conditions: different concentrations of M. avidus in same water temperature and same concentration of M. avidus in different water temperatures. Olive flounder mortality was threshold dependent with both the temperature and M. avidus density parameters. In this paper, we propose a mathematical model to study M. avidus growth in olive flounder based upon the interactions between parasite and host. The mathematical model was logistic growth differential equation (1.2). The parameters were found with Matlab program through the Levenberge-Marquardt method. In theorem, equilibrium values between the infected fish population and dead population could found. Our equilibrium points were a stable equilibrium and an unstable equilibrium. From the equation (1.6), it was possible to predict the amount of cumulative mortality of olive flounder along with the time after M. avidus infection.

• Journal of fish pathology
• /
• v.35 no.1
• /
• pp.113-120
• /
• 2022

Scuticociliatosis caused by Miamiensis avidus is a very important parasitic disease in olive flounder farming industry. The aim of this study was to determine effect of combined treatment with blue LED (light-emitting diode) illumination and formalin on olive flounder (Paralichthys olivaceus) infected with M. avidus. Different intensity of 405 nm LED (20, 40, and 60 μmol·m^-2·s^-1) was illuminated on 2.2×10⁴ cells/well of M. avidus in a 24 well microplate for 24 h. Also, 2.4×10⁴ cells/well of M. avidus were exposed to varying combinations of 60 μmol·m^-2·s^-1 of 405 nm LED and serial 10-fold dilutions of formalin (from 10 to 100 ppm) for 15, 30, 45, and 60 min. Surviving M. avidus were counted using a hemocytometer. For in vivo test, flounder acclimatized at 11-12 practical salinity unit (psu) were challenged with 2×10⁶ cells/ml of M. avidus by immersion method for 1 h. Then, fish were moved and divided into four groups; "F" group, treated with formalin at 50 ppm; "L" group, treated with 60 μmol·m^-2·s^-1 of 405 nm LED; "C" group, treated with combination of the two methods; and the control group. After treatment for 30 min, fish were transferred to new tanks (salinity = 11-12 psu) and observed for 3 weeks. As a result, illumination of 405 nm LED at 60 μmol·m^-2·s^-1 killed 100% of M. avidus after 12 h, while 67% and 90% of the scuticociliate died at 20 and 40 μmol·m^-2·s^-1, respectively, after 24 h exposure. One hundred percent of M. avidus was killed at 90, 80, 80 and 70 ppm after exposure to formalin for 15, 30, 45 and 60 min, respectively. However, combined method (e.g., 60 μmol·m^-2·s^-1 of 405 nm-LED plus 50 ppm formalin) killed the parasite within 30 min. From in vivo test, similarly, survival rates of fish challenged with M. avidus were 100%, 43%, 29% and 0% in the C, F, L, and control groups, respectively. Results obtained in this study demonstrates that the combined treatment method has clear synergistic effect on scuticociliatosis in fish.