In vitro growth requirements for the fish pathogen Philasterides dicentrarchi (Ciliophora, Scuticociliatida)

Iglesias, R.; Paramá, A.; Álvarez, M. F.; Leiro, J.; Aja, C; Sanmartín, M. L.

doi:10.1016/s0304-4017(02)00350-3

Cited by 57 publications

(55 citation statements)

References 23 publications

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“…They are also believed to play key roles in the processes of penetration and migration of parasites through host tissues (Klemba & Goldberg 2002 et al 1972, Cain & Morado 2001. Densities of the scuticociliate fish pathogen Philasterides dicentrarchi have also been observed to multiply 5-fold over 7 d (reaching a plateau phase after 3 d) during in vitro culture using a similar seeding density to that used in the present study (Inglesias et al 2003).Associated with the increase in growth of the ciliate in vitro, was the release of multiple extracellular proteases. As the ciliates were apparently always motile, and were still increasing in number when the experiment was terminated (Day 7), we concluded that the extracellular proteases detected in the culture medium had been actively secreted.…”

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confidence: 61%

Identification and partial characterisation of metalloproteases secreted by a Mesanophrys-like ciliate parasite of the Norway lobster Nephrops norvegicus

Small¹,

Neil²,

Taylor³

et al. 2005

Dis. Aquat. Org.

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A ciliate parasite, tentatively identified as Mesanophrys sp. of Norway lobstersNephrops norvegicus, is demonstrated to secrete several proteases into the culture medium (modified Nephrops saline). Analyses using substrate-impregnated sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed 12 activity bands differing greatly in mobility in the gels. The complete inhibition of proteolytic activity by 1,10-phenanthroline indicated that the proteases are of the metallo class. The proteases were active at the physiological temperature (8°C) and haemolymph pH (7.8) of the host. The proteases were selective in the degradation of several host proteins, including the myosin heavy chain, which is a major structural component of lobster muscle. Consequently, these proteases may have important roles in several aspects of the host -parasite interaction including invasion, nutrient uptake by the ciliate, and pathogenesis.KEY WORDS: Ciliate · Parasite · Proteases · Secreted · Pathology Resale or republication not permitted without written consent of the publisherDis Aquat Org 67: [225][226][227][228][229][230][231] 2005 ( Lee et al. 2003); in this case, secreted proteases may be directly responsible for many of the clinical symptoms associated with the disease, such as tissue necrosis.Several reports of Mesanophrys spp. infections in crustaceans have indicated that these ciliates infiltrate and consume host tissues and haemocytes (Poisson 1930, Bang et al. 1972, Messick & Small 1996. The discovery of a Mesanophrys-like parasitic ciliate infecting Norway lobsters Nephrops norvegicus, and isolation of the ciliate via in vitro cultivation, has allowed proteolytic factors that may play a role in the parasite's establishment and proliferation to be investigated. This paper reports on the characterisation of these proteases, and suggests ways in which they may be involved in the pathogenesis of ciliate disease in lobsters. MATERIALS AND METHODSIsolation and cultivation of the ciliate parasite. Routine screening of Nephros norvegicus from the Clyde Sea area for the parasitic dinoflagellate Hematodinium sp. led to the discovery and identification of a ciliate infection in several Norway lobsters (Small et al. 2005). Haemolymph samples were withdrawn aseptically from the base of the fifth pereiopod using a 1 ml disposable syringe and 25-gauge needle following sterilisation of the cuticle with 70% (v/v) ethanol. Ciliate infections were diagnosed by viewing haemolymph smears using light microscopy. Parasitic ciliates were isolated in 3.5 cm well plates with 0.2 ml infected haemolymph added to 5 ml culture medium in each well. The culture medium comprised 10% (v/v) ) and streptomycin (10 µg ml -1 ) were added to inhibit bacterial contamination. The medium was filter-sterilised (0.2 µm) after the addition of all constituents. Cultures were incubated at 8°C. Serial subculturing gave rise to axenic cultures.Experimental subcultures for protease analyses were initiated with the addition of 1 × 10 5 ciliate...

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confidence: 61%

Identification and partial characterisation of metalloproteases secreted by a Mesanophrys-like ciliate parasite of the Norway lobster Nephrops norvegicus

Small¹,

Neil²,

Taylor³

et al. 2005

Dis. Aquat. Org.

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“…The influence of temperature was demonstrated by Iglesias et al (2003a) for cultured Philasterides dicentrarchi, with optimum growth at 18-23ºC and negligible growth at 13ºC. All the isolates studied in our work grow well at 15ºC (the temperature of routine maintenance), although growth is much faster at 20ºC (Fig.…”

Section: In Vitro Studiesmentioning

confidence: 89%

“…The different isolates of this study have been maintained in axenic culture in this medium for up to 24 months. Iglesias et al (2003a) reported the use of L-15 with 10% FBS for the culture of the ciliate obtained from turbot and identified as Philasterides dicentrarchi. However, they supplemented the medium with glucose, nucleosides, lecithin and Tween 80.…”

Section: In Vitro Studiesmentioning

confidence: 99%

“…Main clinical signs and general histopathology are similar in the different outbreaks, but some differences have been pointed out (Iglesias et al 2001). Apart from the description of the disease, the most complete information available deals with the parasite reported as Philasterides dicentrarchi by Iglesias et al (2001) and includes data on cultivation (Iglesias et al 2003a), antigenicity (Iglesias et al 2003b) and invasion routes of the ciliate (Paramá et al 2003).…”

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confidence: 99%

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Histophagous scuticociliatids (Ciliophora) parasitizing turbot Scophthalmus maximus: morphology, in vitro culture and virulence

Álvarez-Pellitero¹,

Palenzuela²,

Padrós³

et al. 2004

FOLIA PARASIT

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Abstract. Systemic ciliatosis caused by histophagous ciliates constitutes a serious disease of cultured turbot. Six ciliate isolates were obtained from parasitized turbot during six epizootics at four different farms located in Spain, France and Portugal. Axenic cultures of the six isolates were obtained by periodical subculturing in ATCC 1651MA or supplemented L-15 media. In basal media or seawater, the parasites could survive starving for long periods with no apparent proliferation. In adequate media, growth kinetics was found to be very similar for isolates A and B, with a clear influence of temperature. Morphological studies demonstrated that all isolates share common features that allows their assignment to either Philasterides Kahl, 1931 or Miamiensis Thompson et Moewus, 1964. However, statistically significant differences were evident in pairwise comparisons of the isolates from the four farm sites in 16 taxonomically relevant morphometric features. This could allow the discrimination of different species or strains. Virulence of isolates A and B for healthy turbot was tested in several experiments. Differences in the virulence were especially evident after long-term in vitro culturing, isolate A being clearly attenuated after 35-42 passages, whereas isolate B became more virulent after 20-42 passages. The need of further studies to confirm such virulence variability and its implications in pathogenesis and prevention of turbot scuticociliatoses is stressed.Scuticociliates are ciliate protozoans (Ciliophora) occurring abundantly in eutrophic coastal and saprobic maricultural waters. Some of them can behave as opportunistic histophagous parasites causing severe infections in crustaceans and fish. Small (1967) proposed the new order Scuticociliatida including the suborders Pleuronematina, Pseudocohnilembina and Philasterina. Further revisions of the Ciliophora (Lynn and Small 2000) granted scuticociliates the level of subclass (Scuticociliatia), and the suborder Philasterina was ranked as order (Philasterida). Since the criteria used hitherto to classify the families, genera and species of scuticociliates have been variable, the taxonomy of these organisms is currently rather confusing.Recently, different scuticociliates have become an important threat for cultured marine fish such as tuna and different flatfish. Fatal encephalitis of bluefin tuna, attributed to Uronema nigricans, was reported by Munday et al. (1997). Among flatfish, Jee et al. (2001) described scuticociliatosis of Japanese flounder, Paralichthys olivaceus. In turbot, different episodes of scuticociliatosis have been reported in Spain (Dyková and Figueras 1994, Iglesias et al. 2001, authors' unpublished data) and Norway (Sterud et al. 2000). Only in one of these turbot epizootics the causative organism was identified to specific level as Philasterides dicentrarchi Dragesco, Dragesco, Coste, Gasc, Romestand, Raymond et Bouix, 1995by Iglesias et al. (2001. This species was originally described from an episode of Mediterranean seabass (Dice...

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“…Este parámetro ambiental también ha sido relacionado con brotes de la enfermedad en otros peces y crustáceos (Cawthorn 1997, Munday et al 1997, Iglesias et al 2001, Azad 2007. Iglesias et al (2003) reportaron una mayor tasa de crecimiento y reproducción de P. dicentrarchi, a 22 °C que a 15 °C en cultivo. Puig et al (2007) realizaron infecciones experimentales con P. dicentrarchi en el turbot S. maximus y reportaron que el curso de la infección fue rápido a mayor temperatura, concluyendo que este parámetro influye en el desarrollo de los parásitos.…”

Section: Discussionunclassified

Ocurrencia de escuticociliatosis en el lenguado Paralichthys adspersus causado por Miamiensis avidus, en Perú

et al. 2016

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Información sobre los autores:MM y GS diseñaron el estudio, MM colectó las muestras, realizó la descripción histológica, GS realizó el análisis de identificación molecular, VF y CF realizaron el análisis microbiológico y MM, GS, VF y CF redactaron y aprobaron el manuscrito Los autores no incurren en conflictos de intereses. ResumenLa escuticociliatosis es una enfermedad causada por ciliados del orden Scuticociliatida los cuales se caracterizan por su alto potencial para invadir al huésped, significando un serio problema en la acuicultura marina. El presente trabajo describe la infección con escuticociliados en lenguado Paralichthys adspersus en cautiverio. Los signos externos e internos de la infección incluyen zonas necróticas en el tegumento, abundante mucosidad, abultamiento de la cavidad visceral con acumulación de líquido ascítico, necrosis de las fibras musculares, licuefacción del cerebro, entre otros. Se identificó y caracterizó molecularmente al parásito ciliado como M. avidus, utilizando secuencias de gen mitocondrial citocromo oxidasa I (COI), y de los genes nucleares β-tubulina y la región de la subunidad menor rRNA 18S, mostrando una sinonimia con P. dicentrarchi. Todas las lesiones estuvieron invadidas de ciliados. A nivel histológico, se detectaron ciliados en casi todos los tejidos siendo el cerebro el órgano más parasitado. Los ejemplares infectados, acompañados de infecciones bacterianas secundarias predominantemente por Vibrio alginolyticus, después de un periodo de letargia mueren.Palabras clave: escuticociliatosis; Miamiensis avidus; lenguado Paralichthys adpsersus; acuicultura; parásitos ciliados. AbstractThe Scuticociliatosis, a disease caused by ciliates from the order Scuticociliatida characterized by their high potential to invade the host, is a serious problem in marine aquaculture. This paper describes scuticociliatosis infection in farmed flounder Paralichthys adspersus. External and internal signs of infection include necrotic areas in the tegument, abundant mucus, swelling of the visceral cavity with ascitic fluid accumulation, necrotic muscle fibers and brain liquefaction, among others. The ciliate parasite was molecularly identified and characterized as M. avidus, using sequences of mitochondrial gene cytochrome oxidase I (COI), and nuclear genes β-tubulin and the region of the small subunit 18S rRNA, showing synonymy with P. dicentrarchi . All lesions were infested by ciliates. Histologically, ciliates are detected in almost all tissues being the brain the organ more parasitized. The infected specimens associated with secondary bacterial infections, predominantly Vibrio alginolyticus, died after a lethargy period.

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In vitro growth requirements for the fish pathogen Philasterides dicentrarchi (Ciliophora, Scuticociliatida)

Cited by 57 publications

References 23 publications

Identification and partial characterisation of metalloproteases secreted by a Mesanophrys-like ciliate parasite of the Norway lobster Nephrops norvegicus

Identification and partial characterisation of metalloproteases secreted by a Mesanophrys-like ciliate parasite of the Norway lobster Nephrops norvegicus

Histophagous scuticociliatids (Ciliophora) parasitizing turbot Scophthalmus maximus: morphology, in vitro culture and virulence

Ocurrencia de escuticociliatosis en el lenguado Paralichthys adspersus causado por Miamiensis avidus, en Perú

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