DOCSLIB.ORG

Effectiveness of Chemotherapeutants Against Scuticociliate Philasterides Dicentrarchi, a Parasite of Olive flounder

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Effectiveness of Chemotherapeutants Against Scuticociliate Philasterides Dicentrarchi, a Parasite of Olive flounder Veterinary Parasitology 168 (2010) 19–24 Contents lists available at ScienceDirect Veterinary Parasitology journal homepage: www.elsevier.com/locate/vetpar Effectiveness of chemotherapeutants against scuticociliate Philasterides dicentrarchi, a parasite of olive flounder Chang-Nam Jin a,1, Ramasamy Harikrishnan b,1, Young-Gun Moon b, Man-Chul Kim b, Ju-Sang Kim b, Chellam Balasundaram c, Moon-Soo Heo b,* a Jeollanamdo Fisheries Technology Office, Jindo, Jeonnan 539 802, South Korea b Marine Applied Microbes and Aquatic Organism Disease Control Lab, Aquatic Biomedical Sciences, School of Marine Biomedicals Sciences, College of Ocean Science & Marine and Enviromental Research Institute, Jeju National University, Jeju 690 756, South Korea c Department of Animal Science, Bharathidasan University, Tiruchirapalli 620 024, Tamil Nadu, India ARTICLE INFO ABSTRACT Article history: The histophagous scuticociliate Philasterides dicentrarchi causes fatal scuticociliatosis in Received 8 May 2009 farmed olive flounder Paralichthys olivaceus. The average monthly prevalence of Received in revised form 11 September 2009 scuticociliatosis with P. dicentrarchi infections was very high from May to July (40 Æ 3 Accepted 1 October 2009 to 79 Æ 2%) in olive flounder at farms of Jeju Island, South Korea, from 2000 to 2004. The prevalence of mixed infection along with Vibrio spp. infection was higher (49 Æ 8%) than that Keywords: of scuticociliatosis alone. To date no effective control measure for P. dicentrarchi infection has Formalin been described and large economic losses continue. In the present study 3 chemotheraputants Jenoclean Hydrogen peroxide (formalin, hydrogen peroxide and Jenoclean) were used. Among these, bath treatment with Philasterides dicentrarchi Jenoclean at a low concentration of 50 ppm proved effective; the results were confirmed with Paralichthys olivaceus in vitro motility assessments and morphological changes in P. dicentrarchi. A similar trend was noted following hydrogen peroxide treatment. However, formalin was only moderately effective at this concentration. Therefore, hydrogen peroxide and Jenoclean are the promising compounds effective at low concentrations with short application time. ß 2009 Elsevier B.V. All rights reserved. 1. Introduction Scuticociliatosis was first reported in olive flounder in 1990, and quickly became a challenging disease with no The olive flounder Paralichthys olivaceus is an economic- effective control (Chun, 2000). After 2000 the disease ally important food fish in Korea with 98% of the domestic inflicted severe mass mortality in many commercial farms demand met through indigenous farming (Jung et al., in Jeju Island (Oh et al., 2006). Recently, 3 species 2001); farmed production commenced in the 1990s when (Pseudocohnilembus persalinus, Uronema marinum and wild stocks of P. olivaceus were exhausted. Currently olive Philasterides dicentrarchi) have been identified as respon- flounder culture is a major activity with about 300 sible for scuticociliatosis in Korean olive flounder farms commercial farms situated in Jeju Island, South Korea. (Kim et al., 2004a,b) with P. dicentrarchi as the main species Reported annual production was 21,368 and 20,000 to involved (Iglesias et al., 2001; Kim et al., 2004a; Jung et al., 25,000 mt in 1998 and 2000, respectively (Statistical Year 2005). Book of Maritime Affairs and Fisheries, 2000; Korea Flounder culture mainly begins in March and ends in National Statistical Office, 2001). December in a land-based raceway culture system with an average mortality of 15–20% (Lee, 2007). The incidence of the disease begins in May (Jin, 2006) with a marked * Corresponding author. Tel.: +82 64 754 3473; fax: +82 64 756 3493. withering of scales followed by imbalanced swimming E-mail addresses: [email protected] (R. Harikrishnan), behavior, anemia, weight loss, brownish skin patches, [email protected] (M.-S. Heo). 1 These authors contributed equally to this work. necrotic lesions, enteritis, excessive body mucus and 0304-4017/$ – see front matter ß 2009 Elsevier B.V. All rights reserved. doi:10.1016/j.vetpar.2009.10.001 20 C.-N. Jin et al. / Veterinary Parasitology 168 (2010) 19–24 yellowish intestinal mucus. The infected fishes are dip with yeast extract and 10% fetal calf serum (Alvarez-Pellitero treated with Jenoclean, hydrogen peroxide or formalin et al., 2004). Active ciliates were washed and concentrated by (100–500 ppm) for 15–20 min per day for 3–5 days with dilution with Hanks balanced salt solution (HBSS) containing partial success. Indeed only about 50% of the farmers an antibiotic/antimycotic mixture (PSA) at 1Â to 3Â concen- attempt this and others go for final harvest though the final tration (1Â PSA = 100 U mlÀ1 penicillin, 100 mgmlÀ1 strepto- size is less (220 Æ 40 g) in August to October than in mycin and 0.25 mgmlÀ1 amphotericin B), followed by December (440 Æ 100 g). Farmers at present partially manage centrifugation at 500 Â g for 15 min. This process was repeated scuticociliatosis in the farm with crude application of as needed to obtain clear inocula with viable parasites. antibiotics such as oxy-tetracycline, gentamycine, tetracy- cline, amoxycilin and cefazolin, and/or chemicals such as 2.2. Subculture and harvesting of ciliates formalin, hydrogen peroxide, sodium chloride and Genoclean at concentrations of 350 Æ 150 ppm; however, no scientific Sterile artificial sea water (20% salinity) prepared using study has yet been conducted to determine relative efficacy Instant Ocean mixture (Aquarium Systems, Sarreborg, of these treatments. France) was used as a basal medium for the initial primary Globally, P. dicentrarchi poses a serious threat to the cultures. Ciliates were maintained in 25-cm2 T-flasks, with aquaculture industry, causing mass mortalities in marine 7 ml media at 20 8C. Subcultures were initiated by fishes such as olive flounder (Yoshinaga and Nakazoe, harvesting cultures in stationary phase, followed by 1993; Jee et al., 2001; Kwon et al., 2003), sea-bass and centrifugation (500–650 Â g, 15 min) and washing with turbot (Dykova´ and Figueras, 1994; Dragesco et al., 1995; HBSS containing 2Â PSA. Parasites were counted in a Iglesias et al., 2001; Sterud et al., 2000). However, detailed Neubauer chamber and inoculated into fresh media identification of its characteristics and possible treatment (Alvarez-Pellitero et al., 2004). measures are yet to be elucidated. For these reasons, it is essential to screen new effective drugs against P. dicen- 2.3. Determination of the efficacy of chemotherapeutants trarchi infection. Recently in vivo (piperazine N-substituted naphthyridines, pyridothienopyrimidines and pyridothie- Effects were observed using 1.5 ml of ciliate suspension notriazines, Quintela et al., 2003) studies report on the (centrifuged and resuspended in 2 mm filtered salt water) therapeutic properties of antiprotozoal compounds on the with 200 ml of suitably diluted therapeutants to give the control of the disease. In finfish culture, formalin baths are desired final concentration. Ciliate densities for each trial widely applied at concentrations of 35 Æ 15 ppm for 90 min ranged from 1.5 Â 104 to 4.3 Â 104 cells mlÀ1. Compounds (Tonguthai and Chanratchakool, 1992; Scott, 1993; Fajer- tested for efficacy in killing the ciliates were formalin Avila et al., 2003) or for a shorter duration of 60 min at (Merck, Germany) at 50, 100, 200, 300, and 400 ppm, concentrations from 167 to 250 ppm (Poupard, 1978)in hydrogen peroxide (Merck, Germany) at 25, 50, 100, 150, treating protozoan ectoparasites. 200, and 300 ppm and Jenoclean at 50, 100, 200, 300, 400, To the best of our knowledge, there is a paucity of and 500 ppm. Jenoclean was prepared by one-kg comprises studies on the chemotherapy of scuticociliatosis in olive Atacama extract 97% (Zeolites) and citric acid 3%. Sterile flounder. However, antibiotics like amprolium, monensin, polystyrene microplates (IWAKI) with 24 wells with 0.5 ml oxy-tetracycline and pyrimethamine + sulphaquinoxaline of medium per well (surface-to-volume ratio 4 cm2 mlÀ1) (Bayley, 1995) and amprolium, formalin and monesin were used. Each trial was performed in triplicate. After (Novotny et al., 1996) have been tested in vitro against inoculating each well with P. dicentrarchi, the plates were Anophryoides haernophil. The aim of this study is to assess incubated at 19 Æ 1 8C(Crosbie and Munday, 1999). Effects of the in vitro efficacy of several chemotherapeutic com- therapeutants on motility were observed periodically by pounds (formalin, hydrogen peroxide and Jenoclean) scanning several fields of view at 100Â and 200Â magnifica- against scuticociliatosis caused by P. dicentrarchi. tion using an inverted microscope (Zeiss Invertoscope ID 02) 2. Materials and methods Table 1 Scoring system for appraising the effects of various compounds on motility and morphology of Philasterides dicentrarchi (Crosbie and Munday, 1999). 2.1. Isolation of ciliates Score Interpretation P. olivaceus (mean weight 20 5 g; mean length Æ Motility 13 Æ 2 cm; n = 150) were collected from 15 local land-based 4 No effect; motility normal flounder farms affected with scuticociliatosis located in 3 Motility slowed in more than 50% of ciliates Sambo, Gujwa-eup, west of Jeju Island (338240600N, 2 Approximately 50% of ciliates stationary, but cilia 12683204600E), South Korea during May to September 2005. still beating 1 More than 50% of ciliates stationary, but cilia still The affected fish showed varying levels of tissue damage beating including severe epidermal and dermal necrotic lesions (Jin, 0 No sign of motility or cilia movement 2006). P. dicentrarchi were isolated from the brain, gill, ascites Morphology or ulcerated skin of the collected samples. We cloned and 4 No change; cells elliptical identified P. dicentrarchi using species-specific primers (Kim 3 Less than 50% of cells round or irregular shape et al., 2004b), ultrastructural and phylogenetic studies 2 Approximately 50% of cells round or irregular shape (Parama et al., 2006). P. dicentrarchi were cultured in 1 More than 50% of cells round and lysis evident 0 Extensive lysis with few cells intact minimum essential medium (MEM, Sigma) supplemented C.-N. Jin et al.
Load more

Recommended publications
  • Morphology and Phylogeny of Four Marine Scuticociliates (Protista, Ciliophora), with Descriptions of Two New Species: Pleuronema Elegans Spec
    Acta Protozool. (2015) 54: 31–43 www.ejournals.eu/Acta-Protozoologica ACTA doi:10.4467/16890027AP.15.003.2190 PROTOZOOLOGICA Morphology and Phylogeny of Four Marine Scuticociliates (Protista, Ciliophora), with Descriptions of Two New Species: Pleuronema elegans spec. nov. and Uronema orientalis spec. nov. Xuming PAN1†, Jie HUANG2†, Xinpeng FAN3, Honggang MA1, Khaled A. S. AL-RASHEID4, Miao MIAO5 and Feng GAO1 1Laboratory of Protozoology, Institute of Evolution and Marine Biodiversity, Ocean University of China, Qingdao 266003, China; 2Key Laboratory of Aquatic Biodiversity and Conservation of Chinese Academy of Science, Institute of Hydrobiology, Chinese Academy of Science, Wuhan 430072, China; 3School of Life Sciences, East China Normal University, Shanghai 200062, China; 4Zoology Department, King Saud University, Riyadh 11451, Saudi Arabia; 5College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China †Contributed equally Abstract. The morphology and infraciliature of four marine scuticociliates, Pleuronema elegans spec. nov., P. setigerum Calkins, 1902, P. gro- lierei Wang et al., 2008 and Uronema orientalis spec. nov., collected from China seas, were investigated through live observation and protargol staining methods. Pleuronema elegans spec. nov. can be recognized by the combination of the following characters: size in vivo 90–115 × 45–60 µm, slender oval in outline with a distinctly pointed posterior end; about 10 prolonged caudal cilia; consistently two preoral kineties and 18 or 19 somatic kineties; membranelle 2a double-rowed with its posterior end straight; membranelle 3 three-rowed; one macronucleus; marine habitat. Uronema orientalis spec. nov. is distinguished by the following features: in vivo about 40–55 × 20–30 μm with a truncated apical plate; consis- tently twenty somatic kineties; membranelle 1 single-rowed and divided into two parts which comprise four and three basal bodies respectively; contractile vacuole pore positioned at the end of the second somatic kinety; marine habitat.
    [Show full text]
  • Interactions Between the Parasite Philasterides Dicentrarchi and the Immune System of the Turbot Scophthalmus Maximus.A Transcriptomic Analysis
    biology Article Interactions between the Parasite Philasterides dicentrarchi and the Immune System of the Turbot Scophthalmus maximus.A Transcriptomic Analysis Alejandra Valle 1 , José Manuel Leiro 2 , Patricia Pereiro 3 , Antonio Figueras 3 , Beatriz Novoa 3, Ron P. H. Dirks 4 and Jesús Lamas 1,* 1 Department of Fundamental Biology, Institute of Aquaculture, Campus Vida, University of Santiago de Compostela, 15782 Santiago de Compostela, Spain; [email protected] 2 Department of Microbiology and Parasitology, Laboratory of Parasitology, Institute of Research on Chemical and Biological Analysis, Campus Vida, University of Santiago de Compostela, 15782 Santiago de Compostela, Spain; [email protected] 3 Institute of Marine Research, Consejo Superior de Investigaciones Científicas-CSIC, 36208 Vigo, Spain; [email protected] (P.P.); antoniofi[email protected] (A.F.); [email protected] (B.N.) 4 Future Genomics Technologies, Leiden BioScience Park, 2333 BE Leiden, The Netherlands; [email protected] * Correspondence: [email protected]; Tel.: +34-88-181-6951; Fax: +34-88-159-6904 Received: 4 September 2020; Accepted: 14 October 2020; Published: 15 October 2020 Simple Summary: Philasterides dicentrarchi is a free-living ciliate that causes high mortality in marine cultured fish, particularly flatfish, and in fish kept in aquaria. At present, there is still no clear picture of what makes this ciliate a fish pathogen and what makes fish resistant to this ciliate. In the present study, we used transcriptomic techniques to evaluate the interactions between P. dicentrarchi and turbot leucocytes during the early stages of infection. The findings enabled us to identify some parasite genes/proteins that may be involved in virulence and host resistance, some of which may be good candidates for inclusion in fish vaccines.
    [Show full text]
  • Disease of Aquatic Organisms 86:163
    Vol. 86: 163–167, 2009 DISEASES OF AQUATIC ORGANISMS Published September 23 doi: 10.3354/dao02113 Dis Aquat Org NOTE DNA identification of ciliates associated with disease outbreaks in a New Zealand marine fish hatchery 1, 1 1 1 2 P. J. Smith *, S. M. McVeagh , D. Hulston , S. A. Anderson , Y. Gublin 1National Institute of Water and Atmospheric Research (NIWA), Private Bag 14901, Wellington, New Zealand 2NIWA, Station Road, Ruakaka, Northland 0166, New Zealand ABSTRACT: Ciliates associated with fish mortalities in a New Zealand hatchery were identified by DNA sequencing of the small subunit ribosomal RNA gene (SSU rRNA). Tissue samples were taken from lesions and gill tissues on freshly dead juvenile groper, brain tissue from adult kingfish, and from ciliate cultures and rotifers derived from fish mortality events between January 2007 and March 2009. Different mortality events were characterized by either of 2 ciliate species, Uronema marinum and Miamiensis avidus. A third ciliate, Mesanophrys carcini, was identified in rotifers used as food for fish larvae. Sequencing part of the SSU rRNA provided a rapid tool for the identification and mon- itoring of scuticociliates in the hatchery and allowed the first identification of these species in farmed fish in New Zealand. KEY WORDS: Small subunit ribosomal RNA gene · Scuticociliatosis · Uronema marinum · Miamiensis avidus · Mesanophrys carcini · Groper · Polyprion oxygeneios · Kingfish · Seriola lalandi Resale or republication not permitted without written consent of the publisher INTRODUCTION of ciliate pathogens in fin-fish farms (Kim et al. 2004a,b, Jung et al. 2007) and in crustacea (Ragan et The scuticociliates are major pathogens in marine al.
    [Show full text]
  • Protozoologica Acta Doi:10.4467/16890027AP.15.027.3541 Protozoologica
    Acta Protozool. (2015) 54: 325–330 www.ejournals.eu/Acta-Protozoologica ActA doi:10.4467/16890027AP.15.027.3541 Protozoologica Short Communication High-Density Cultivation of the Marine Ciliate Uronema marinum (Ciliophora, Oligohymenophorea) in Axenic Medium Weibo ZHENG1, Feng GAO1, Alan WARREN2 1 Laboratory of Protozoology, Institute of Evolution and Marine Biodiversity, Ocean University of China, Qingdao 266003, China; 2 Department of Life Sciences, Natural History Museum, London SW7 5BD, UK Abstract. Uronema marinum is a cosmopolitan marine ciliate. It is a facultative parasite and the main causative agent of outbreaks of scuticociliatosis in aquaculture fish. This study reports a method for the axenic cultivation of U. marinum in high densities in an artificial medium comprising proteose peptone, glucose and yeast extract powder as its basic components. The absence of bacteria in the cultures was confirmed by fluorescence microscopy of DAPI-stained samples and the failure to recover bacterial SSU-rDNA using standard PCR methods. Using this axenic medium, a maximum cell density of 420,000 ciliate cells/ml was achieved, which is significantly higher than in cultures using living bacteria as food or in other axenic media reported previously. This method for high-density axenic cultivation of U. marinum should facilitate future research on this economically important facultative fish parasite. Key words: Axenic cultivation, ciliates, fish parasite, scuticociliatosis, Uronema marinum. INTRODUCTION fully cultivated axenically and was grown in a medium containing dead yeast cells, liver extract and kidney tis- sue (Glaser et al. 1933). Pure cultures of strains belong- Axenic cultures of ciliates have proved to be ex- ing to the “Colpidium-Glaucoma-Leucophrys-Tetrahy- tremely valuable in a wide range of research fields mena group” were subsequently established (Corliss including growth, nutrition, respiration, genetics, fac- 1952).
    [Show full text]
  • Miamiensis Avidus
    Parasitology Research https://doi.org/10.1007/s00436-018-6010-8 ORIGINAL PAPER Development of a safe antiparasitic against scuticociliates (Miamiensis avidus) in olive flounders: new approach to reduce the toxicity of mebendazole by material remediation technology using full-overlapped gravitational field energy Jung-Soo Seo1 & Na-Young Kim2 & Eun-Ji Jeon2 & Nam-Sil Lee2 & En-Hye Lee3 & Myoung-Sug Kim2 & Hak-Je Kim4 & Sung-Hee Jung2 Received: 5 March 2018 /Accepted: 6 July 2018 # The Author(s) 2018 Abstract The olive flounder (Paralychthys olivaceus) is a representative farmed fish species in South Korea, which is cultured in land-based tanks and accounts for approximately 50% of total fish farming production. However, farmed olive flounder are susceptible to infection with parasitic scuticociliates, which cause scuticociliatosis, a disease resulting in severe economic losses. Thus, there has been a longstanding imperative to develop a highly stable and effective antiparasitic drug that can be rapidly administered, both orally and by bath, upon infection with scuticociliates. Although the efficacy of commercially available mebendazole (MBZ) has previously been established, this compound cannot be used for olive flounder due to hematological, biochemical, and histopathological side effects. Thus, we produced material remediated mebendazole (MR MBZ), in which elements comprising the molecule wereARTICLE remediated by using full-overlapped grav- itational field energy, thereby reducing the toxicity of the parent material. The antiparasitic effect of MR MBZ against scuticociliates in olive flounder was either similar to or higher than that of MBZ under the same conditions. Oral (100 and500mg/kgB.W.)andbath(100and500mg/L) administrations of MBZ significantly (p < 0.05) increased the values of hematological and biochemical parameters, whereas these values showed no increase in the MR MBZ administration group.
    [Show full text]
  • Biodiversity of Marine Scuticociliates (Protozoa, Ciliophora) from China
    Available online at www.sciencedirect.com ScienceDirect European Journal of Protistology 51 (2015) 142–157 Biodiversity of marine scuticociliates (Protozoa, Ciliophora) from China: Description of seven morphotypes including a new species, Philaster sinensis spec. nov. a,b a,∗ a b Xuming Pan , Zhenzhen Yi , Jiqiu Li , Honggang Ma , c c Saleh A. Al-Farraj , Khaled A.S. Al-Rasheid a Key Laboratory of Ecology and Environmental Science in Guangdong Higher Education, South China Normal University, Guangzhou 510631, China b Laboratory of Protozoology, Institute of Evolution & Marine Biodiversity, Ocean University of China, Qingdao 266003, China c Zoology Department, King Saud University, Riyadh 11451, Saudi Arabia Received 21 August 2014; received in revised form 17 February 2015; accepted 18 February 2015 Available online 25 February 2015 Abstract Seven marine scuticociliates, Philaster sinensis spec. nov., Pseudocohnilembus hargisi Evans and Thompson, 1964. J. Pro- tozool. 11, 344, Parauronema virginianum Thompson, 1967. J. Protozool. 14, 731, Uronemella filificum (Kahl, 1931. Tierwelt. Dtl. 21, 181) Song and Wilbert, 2002. Zool. Anz. 241, 317, Cohnilembus verminus Kahl, 1931, Parauronema longum Song, 1995. J. Ocean Univ. China. 25, 461 and Glauconema trihymene Thompson, 1966. J. Protozool. 13, 393, collected from Chinese coastal waters, were investigated using live observations, silver impregnation methods, and, in the case of the new species, SSU rDNA sequencing. Philaster sinensis spec. nov. can be recognized by the combination of the following characters: body cylindrical, approximately 130–150 × 35–55 ␮m in vivo; apical end slightly to distinctly pointed, posterior generally rounded; 19–22 somatic kineties; M1 triangular, consisting of 13 or 14 transverse rows of kinetosomes; M2 comprising 10–12 longitudi- nal rows; CVP positioned at end of SK1; marine habitat.
    [Show full text]
  • Systematic Studies on Ciliates (Alveolata, Ciliophora) in China: Progress
    Available online at www.sciencedirect.com ScienceDirect European Journal of Protistology 61 (2017) 409–423 Review Systematic studies on ciliates (Alveolata, Ciliophora) in China: Progress and achievements based on molecular information a,1 a,b,1 a,c,1 a,d,1 a,e,1 Feng Gao , Jie Huang , Yan Zhao , Lifang Li , Weiwei Liu , a,f,1 a,g,1 a,1 a,h,∗ Miao Miao , Qianqian Zhang , Jiamei Li , Zhenzhen Yi , i j a,k Hamed A. El-Serehy , Alan Warren , Weibo Song a Institute of Evolution and Marine Biodiversity, Ocean University of China, Qingdao 266003, China b Key Laboratory of Aquatic Biodiversity and Conservation, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China c Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China d Marine College, Shandong University, Weihai 264209, China e Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Science, Guangzhou 510301, China f College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China g Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai 264003, China h Guangzhou Key Laboratory of Subtropical Biodiversity and Biomonitoring, South China Normal University, Guangzhou 510631, China i Department of Zoology, King Saud University, Riyadh 11451, Saudi Arabia j Department of Life Sciences, Natural History Museum, London SW7 5BD, UK k Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266003, China Available online 6 May 2017 Abstract Due to complex morphological and convergent morphogenetic characters, the systematics of ciliates has long been ambiguous.
    [Show full text]
  • Miamiensis Avidus (Ciliophora: Scuticociliatida) Causes Systemic
    DISEASES OF AQUATIC ORGANISMS Vol. 73: 227–234, 2007 Published January 18 Dis Aquat Org Miamiensis avidus (Ciliophora: Scuticociliatida) causes systemic infection of olive flounder Paralichthys olivaceus and is a senior synonym of Philasterides dicentrarchi Sung-Ju Jung*, Shin-Ichi Kitamura, Jun-Young Song, Myung-Joo Oh Department of Aqualife Medicine, Chonnam National University, Chonnam 550-749, Korea ABSTRACT: The scuticociliate Miamiensis avidus was isolated from olive flounder Paralichthys oli- vaceus showing typical symptoms of ulceration and hemorrhages in skeletal muscle and fins. In an infection experiment, olive flounder (mean length: 14.9 cm; mean weight: 26.8 g) were immersion challenged with 2.0 × 103, 2.0 × 104 and 2.0 × 105 ciliates ml–1 of the cloned YS1 strain of M. avidus. Cumulative mortalities were 85% in the 2.0 × 103 cells ml–1 treatment group and 100% in the other 2 infection groups. Many ciliates, containing red blood cells in the cytoplasm, were observed in the gills, skeletal muscle, skin, fins and brains of infected fish, which showed accompanying hemorrhagic and necrotic lesions. Ciliates were also observed in the lamina propria of the digestive tract, pharynx and cornea. The fixed ciliates were 31.5 ± 3.87 µm in length and 18.5 ± 3.04 µm in width, and were ovoid and slightly elongated in shape, with a pointed anterior and a rounded posterior, presenting a caudal cilium. Other morphological characteristics were as follows: 13 to 14 somatic kineties, oral cil- iature comprising membranelles M1, M2, M3, and paroral membranes PM1 and PM2, contractile vacuole at the posterior end of kinety 2, shortened last somatic kinety and a buccal field to body length ratio of 0.47 ± 0.03.
    [Show full text]
  • Identification and Molecular Characterization Of
    www.nature.com/scientificreports OPEN Identifcation and Molecular Characterization of Superoxide Dismutases Isolated From Received: 19 June 2019 Accepted: 22 August 2019 A Scuticociliate Parasite: Published: xx xx xxxx Physiological Role in Oxidative Stress Iria Folgueira1, Jesús Lamas2, Ana Paula de Felipe1, Rosa Ana Sueiro1 & José Manuel Leiro 2 Philasterides dicentrarchi is a free-living microaerophilic scuticociliate that can become a facultative parasite and cause a serious parasitic disease in farmed fsh. Both the free-living and parasitic forms of this scuticociliate are exposed to oxidative stress associated with environmental factors and the host immune system. The reactive oxygen species (ROS) generated by the host are neutralized by the ciliate by means of antioxidant defences. In this study we aimed to identify metalloenzymes with superoxide − dismutase (SOD) activity capable of inactivating the superoxide anion (•O2 ) generated during induction of oxidative stress. P. dicentrarchi possesses the three characteristic types of SOD isoenzymes in eukaryotes: copper/zinc-SOD, manganese-SOD and iron-SOD. The Cu/Zn-SOD isoenzymes comprise three types of homodimeric proteins (CSD1-3) of molecular weight (MW) 34–44 kDa and with very diferent AA sequences. All Cu/Zn-SODs are sensitive to NaCN, located in the cytosol and in the alveolar sacs, and one of them (CSD2) is extracellular. Mn- and Fe-SOD transcripts encode homodimeric proteins (MSD and FSD, respectively) in their native state: a) MSD (MW 50 kDa) is insensitive to H2O2 and NaN3 and is located in the mitochondria; and b) FSD (MW 60 kDa) is sensitive to H2O2, NaN3 and the polyphenol trans-resveratrol and is located extracellularly.
    [Show full text]
  • Metagenomic Next-Generation Sequencing Reveals Miamiensis Avidus (Ciliophora: Scuticociliatida)
    bioRxiv preprint doi: https://doi.org/10.1101/301556; this version posted April 15, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Retallack et al., mNGS reveals M. avidus in epizootic of leopard sharks 1 Metagenomic next-generation sequencing reveals Miamiensis avidus (Ciliophora: Scuticociliatida) in the 2017 epizootic of leopard sharks (Triakis semifasciata) in San Francisco Bay, California Hanna Retallack1, Mark S. Okihiro*2, Elliot Britton3, Sean Van Sommeran4, Joseph L. DeRisi1,5 1 Department of Biochemistry and Biophysics, University of California San Francisco, 1700 4th St., San Francisco, CA 94158 2 Fisheries Branch, Wildlife and Fisheries Division, California Department of Fish and Wildlife, 1880 Timber Trail, Vista, CA 92081 3 San Francisco University High School, 3065 Jackson St., San Francisco CA 94115 4 Pelagic Shark Research Foundation, 750 Bay Ave. #2108, Capitola CA 95010 5 Chan-Zuckerberg Biohub, 499 Illinois St., San Francisco, CA 94158 *Corresponding author: Mark S. Okihiro California Department of Fish and Wildlife Wildlife and Fisheries Division, Fisheries Branch 1880 Timber Trail Vista, California 92081 Phone: (760) 310-4212 Email: [email protected] Word count: 4042 bioRxiv preprint doi: https://doi.org/10.1101/301556; this version posted April 15, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.
    [Show full text]
  • Further Analyses on the Phylogeny of the Subclass Scuticociliatia (Protozoa, Ciliophora) Based on Both Nuclear and Mitochondrial
    Molecular Phylogenetics and Evolution 139 (2019) 106565 Contents lists available at ScienceDirect Molecular Phylogenetics and Evolution journal homepage: www.elsevier.com/locate/ympev Further analyses on the phylogeny of the subclass Scuticociliatia (Protozoa, Ciliophora) based on both nuclear and mitochondrial data T ⁎ Tengteng Zhanga,b,1, Xinpeng Fanc,1, Feng Gaoa,b, , Saleh A. Al-Farrajd, Hamed A. El-Serehyd, ⁎ Weibo Songa,b,e, a Institute of Evolution & Marine Biodiversity, Ocean University of China, Qingdao 266003, China b Key Laboratory of Mariculture (Ocean University of China), Ministry of Education, Qingdao 266003, China c School of Life Sciences, East China Normal University, Shanghai 200241 China d Zoology Department, College of Science, King Saud University, Riyadh 11451, Saudi Arabia e Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266003, China ARTICLE INFO ABSTRACT Keywords: So far, the phylogenetic studies on ciliated protists have mainly based on single locus, the nuclear ribosomal Scuticociliatia DNA (rDNA). In order to avoid the limitations of single gene/genome trees and to add more data to systematic COI analyses, information from mitochondrial DNA sequence has been increasingly used in different lineages of mtSSU-rDNA ciliates. The systematic relationships in the subclass Scuticociliatia are extremely confused and largely un- nSSU-rDNA resolved based on nuclear genes. In the present study, we have characterized 72 new sequences, including 40 Phylogeny mitochondrial cytochrome oxidase c subunit I (COI) sequences, 29 mitochondrial small subunit ribosomal DNA Secondary structure (mtSSU-rDNA) sequences and three nuclear small subunit ribosomal DNA (nSSU-rDNA) sequences from 47 isolates of 44 morphospecies.
    [Show full text]
  • Phylogenetic Relationship
    ABSTRACT Title of Document: PHYLOGENETIC RELATIONSHIP AMONG POLYMORPHIC OLIGOHYMENOPHOREAN CILIATES, WITH GENE EXPRESSION IN LIFE-HISTORY STAGES OF MIAMIENSIS AVIDUS (CILIOPHORA, OLIGOHYMENOPHOREA) Glenn Frederick Gebler Doctor of Philosophy, 2007 Directed By: Dr. Eugene B. Small Department of Biology The Class Oligohymenophorea is a monophyletic group possessing polymorphic taxa. Thus far, relationships within subclasses of oligohymenophorean ciliates and between polymorphic taxa within families are not well resolved. Here, nuclear small subunit rRNA (SSU rRNA) gene sequences from 63 representative taxa, including several polymorphic species, were used to construct phylogenies and test monophyly of the subclass Scuticociliatia and of the polymorphic taxa within the Oligohymenophorea. In addition, suppression subtraction hybridization (SSH) was used to test the hypothesis that genes are differentially expressed during microstome- to-macrostome and tomite-to-microstome transformation in the polymorphic scuticociliate Miamiensis avidus . Phylogenetic analyses confirmed monophyly of the subclasses Peritrichia and Hymenostomatia. The monophyletic scuticociliates encompassed most, but not all, taxa included in this study. The conditional acceptance of the hypothesis supporting monophyly of the Scuticociliatia was due to the ambiguous placement of three taxa, the apostome Anoplophrya marylandensis , the scuticociliate Dexitrichides pangi , and the peniculine Urocentrum turbo . The polymorphic trait most likely arose on at least four, and perhaps on as many as six, separate occasions within the oligohymenophorean ciliates. Several genes previously implicated in morphogenetic processes in eukaryotes were upregulated during microstome-to-macrostome transformation in M. avidus . Those genes were, elongation factor-1 alpha ( Ef-1α), Constans, Constans-like TOC1 (CCT) transcription factor, a disulfide isomerase, heat shock protein 70, step II splicing factor ( Slu7 ), U1 zinc finger protein, and WD40-16 repeat protein.
    [Show full text]