Multifiliis Theronts Research Articles

Immune-relevant genes expressed in rainbow trout following immunisation with a live vaccine against Ichthyophthirius multifiliis

Rainbow trout Oncorhynchus mykiss were immunised by intra-peritoneal injection using a live vaccine based on Ichthyophthirius multifiliis (Ich) theronts, which previously has shown protection against white spot disease. Samples were taken pre-vaccination and on Day 1, 7, 21 and 28 post-immunisation (p.i.). Expression of immune relevant genes in the liver, spleen and head kidney was monitored by qPCR. To describe the immune reaction following this immunisation, a series of genes encoding cytokines, complement factors, immunoglobulins and acute phase reactants were studied. Genes encoding acute phase reactants in the liver were up-regulated with serum amyloid A (SAA) as the most pronounced with a 2299-fold increase at 24 h p.i. Hepcidin and pre-cerebellin were also up-regulated in the liver 24 h p.i., by 7- and 4-fold, respectively. Complement factors C3, C5 and factor B (Bf) were up-regulated in the spleen and the head kidney 24 h and 28 d p.i. Genes encoding immunoglobulins were not up-regulated, but a specific low titer IgM response (titer 25) against parasite antigens was detected by a modified ELISA 4 wk p.i.

Apoptosis in Ichthyophthirius multifiliis is associated with expression of the Fas receptor of theronts

The expression of type I membrane Fas receptors on the surface of Ichthyophthirius multifiliis (Ich) theronts and the possible association between Fas expression and theront apoptosis induced by the immune antibody was examined. Fas receptors were detected on the theront surface using fluorescein isothiocyanate-conjugated mouse monoclonal antibody against Fas. Fas-positive theronts significantly increased with time during in vitro incubation and with increasing theront concentration. Furthermore, the immune cutaneous antibody induced theront apoptosis; however, Fas ligand did not. A highly significant correlation was noted between theront Fas expression and immune cutaneous antibody-induced theront apoptosis. Numbers of apoptotic theronts increased with increasing number of Fas-positive theronts. The data indicated that theront apoptosis induced by immune cutaneous antibody appears to be positively correlated with the expression of Fas on the surface of Ich theronts.

Induced cross‐protection in channel catfish, Ictalurus punctatus (Rafinesque), against different immobilization serotypes of Ichthyophthirius multifiliis

Abstract Channel catfish, Ictalurus punctatus (Rafinesque), were immunized with Ichthyophthirius multifiliis (Ich) theronts and trophonts, and the immune response and host protection against both homologous and heterologous serotypes of Ich were evaluated. Immunizations were done with two immobilization serotypes (ARS4 and ARS6) of live theronts by bath immersion (trial I) and with sonicated trophonts by intraperitoneal (i.p.) injection (trial II). Cutaneous and serum antibody titres against Ich following immunization were measured and survival of catfish was determined after theront challenge. Theronts were immobilized by the antiserum from fish immunized with homologous theronts or trophonts, but not by the serum of fish immunized with the heterologous serotype. Serum from fish immunized by immersion with live theronts showed higher enzyme-linked immunosorbent assay titres against both homologous and heterologous serotypes than fish immunized by i.p. injection of trophonts. Channel catfish immunized by immersion with live theronts or by i.p. injection with sonicated trophonts developed an immune response against Ich and provided cross-protection against challenge from both serotypes (ARS4 and ARS6) of the parasite. Sonicated trophont antigens in aqueous solution by i.p. injection could stimulate an immune response in fish, but the immunity was of short duration.

Cadmium effects on Ichthyophthirius: evidence for metal-sequestration in fish tissues following administration of recombinant vaccines.

We are developing Tetrahymena thermophila as a delivery system for recombinant vaccines against parasitic protozoa, including the common fish parasite, Ichthyophthirius multifiliis. T. thermophila cell lines expressing I. multifiliis genes under the control of a cadmium-inducible metallothionein gene promoter conferred strong protection against a lethal parasite challenge when administered parenterally to naive fish. Nevertheless, given that heavy metals can be toxic to parasites, a question arose as to whether protection resulted from Cd residues carried over with the vaccine, rather than acquired immunity per se. To address this issue, we examined the sensitivity of I. multifiliis to Cd in vitro and determined Cd concentrations in different host tissues following i.p. injection of juvenile channel catfish with the recombinant vaccine. We found that CdCl2 at concentrations > or = 50 ppb were lethal to I. multifiliis theronts in vitro. Furthermore, Cd concentrations were clearly elevated in fish tissues and reached levels equivalent to 74 ng/g wet weight (74 ppb) in the skin within 14 days of injection with recombinant T. thermophila. Nevertheless, fish injected with non-transformed Tetrahymena grown in the presence or absence of CdCl2 showed no significant difference in either relative survival or parasite load following direct challenge with I. multifiliis.

Prevention of an Initial Infestation ofIchthyophthirius multifiliisin Channel Catfish and Blue Tilapia by Potassium Permanganate Treatment

Abstract Potassium permanganate (KMnO4) has been used to control infestations of Ichthyophthirius multifiliis, but its effectiveness has not been reported from controlled efficacy studies. The purpose of this study was to determine the acute toxicity of KMnO4 to the I. multifiliis theront and the concentration needed to prevent an initial infestation of I. multifiliis in juvenile channel catfish Ictalurus punctatus and blue tilapia Tilapia aurea. Ichthyophthirius multifiliis theronts were exposed to concentrations of KMnO4 in 100 μL of well water in 96-well plates and observed for 4 h to determine the acute toxicity. A concentration of 0.9 mg KMnO4/L caused greater than 95% mortality of the theronts in 4 h in well water; the 4-h LC50 (concentration lethal to 50% of test animals) value was estimated to be 0.77 mg/L. Juvenile channel catfish were exposed to 10,000 theronts/L of well water and immediately treated with a single dose of KMnO4. Infestation occurred in controls 6 d after exposure. The lowest eff...

Occurrence of Ichthyophthirius multifiliis within the peritoneal cavities of infected channel catfish Ictalurus punctatus.

Ichthyophthirius multifiliis is a ciliated protozoan parasite that infects the skin and gills of freshwater fish. This report describes the unusual finding of I. multifiliis within the peritoneal cavities of experimentally infected channel catfish Ictalurus punctatus. Twenty catfish fingerlings were exposed to I. multifiliis theronts using a standardized protocol. Five infected fish and 2 control fish were killed at various time points after infection and their tissues examined. Formalin-fixed, paraffin embedded sections were processed for light microscopy and immunohistochemical detection of I. multifiliis immobilization antigen. Trophonts were observed in skin and gill sections of all exposed fish. Parasites were associated with epithelial hyperplasia, focal areas of cellular disruption and necrosis. In addition to these usual sites of infection, individual trophonts were unexpectedly found within the peritoneal cavities of 4 fish. Staining for parasite antigen facilitated their detection within abdominal adipose tissue or adjacent to intestines. This discovery is interesting as it suggests I. multifiliis may be found in tissues other than the skin and gills during the course of a normal infection.

Values for Selected Serum Analytes during ExperimentalIchthyophthirius multifiliisInfection of Channel Catfish

Channel catfish Ictalurus punctatus were exposed to a sublethal dose of infectious Ichthyophthirius multifiliis theronts, and eight serum analytes were measured. Sequential serum samples from each fish were collected for serum immediately before exposure, 4 d postexposure (PE), and 11 d PE. All fish developed moderately heavy infections with the parasite as indicated by the development of cutaneous lesions 5 d PE to infective theronts. Small but statistically significant changes in total protein, creatinine, and alkaline phosphatase values occurred after acute infection with I. multifiliis. Serum concentrations of glucose, albumin, urea nitrogen, alanine aminotransferase, and aspartate aminotransferase remained constant, indicating little disturbance of liver or kidney functions detectable by a serum biochemical profile established during an acute infection with I. multifiliis.