Biomedical subjects

J Leiro

Publications and source records attributed to J Leiro.

At least 19 recordsLinked to original sources

PubMed2006

Scuticociliate proteinases may modulate turbot immune response by inducing apoptosis in pronephric leucocytes.

The role of proteinases of the histiophagous ciliate Philasterides dicentrarchi, purified by affinity chromatography in bacitracin-Sepharose, on apoptosis (programmed cell death) of turbot pronephric leucocytes (PL) was investigated. The results showed that more than 90% of proteinases purified by bacitracin-Sepharose were cysteine proteinases, which lacked significant caspase-3-like activity and generated three main gelatinolytic bands of molecular weights 36, 45 and 77 kDa as determined by gelatine-SDS-PAGE and immunoblot. Viability of PL cells after 24 h stimulation with P. dicentrarchi cysteine proteinases did not differ from that of non-stimulated cells. Apoptosis was confirmed by: (i) caspase activity, (ii) DNA fragmentation, and (iii) nucleus fragmentation. The caspase-3-like activity in PL incubated for 4h in the presence of 125, 250 and 500 microg/ml of proteinases increased in a dose-dependent fashion. The PL DNA was fragmented following 24-h exposure to P. dicentrarchi cysteine proteinases and characteristic DNA ladders consisting of multimers of approximately 180-200 pb were produced. Morphological changes, such as chromatin condensation and nucleus fragmentation, were observed under fluorescence microscopy after DAPI staining of the PL cells incubated with cysteine proteinase-incubated for 24 h. The results suggest that the pathogenic scuticociliate P. dicentrarchi may induce host leucocyte programmed cell death via the production of cysteine proteinases, as a mechanism of pathogenesis and evasion of the turbot innate immune response.

Animals↗

PubMed2006

Ultrastructure and phylogeny of Philasterides dicentrarchi (Ciliophora, Scuticociliatia) from farmed turbot in NW Spain.

Several species of opportunistic histophagous scuticociliates have been implicated in systemic infections of farmed fish. In turbot, scuticociliatosis is an emerging disease, and the identification of the parasite species involved is controversial. We have previously isolated Philasterides dicentrarchi from farmed turbot scuticociliatosis outbreaks in northwest Spain. In the present study, we report detailed ultrastructural studies of this parasite, and investigate phylogenetic relations with other members of the order Philasterida on the basis of sequence comparison of the small-subunit rRNA (SSUrRNA) gene. Ultrastructural study indicates the presence of dikinetids in the anterior two-thirds of the body; micronucleus closely associated with the macronucleus, though not physically connected; numerous mitochondria located below the cell cortex, parallel to the surface; numerous spherical and fusiform extrusomes located close to the plasma membrane. We consider that these characteristics are useful for diagnosis of infections by this parasite. A nested 350-bp nucleotide sequence of the SSUrRNA gene of the turbot P. dicentrachi isolate showed high identity with previously reported SSUrRNA gene sequences from 2 scuticociliates isolated from olive flounder Paralichthys olivaceus in Korea, namely P. dicentrarchi (98%) and Miamiensis avidus (99%); conversely, our P. dicentrarchi sequence showed low identity (86%) with that of Uronema marinum, a scuticociliate that has also been implicated in scuticociliatosis outbreaks in turbot in Europe and olive flounder in Asia. Phylogenetic tree construction on the basis of the SSUrRNA gene sequences, using the neighbour-joining method, confirm that the different P. dicentrarchi isolates and M. avidus are closely related and a possible synonymy between both ciliates species should be considered.

Animals↗

PubMed2005

In vitro efficacy of glutaraldehyde-crosslinked chitosan microspheres against the fish-pathogenic ciliate Philasterides dicentrarchi.

Philasterides dicentrarchi is a protozoan ciliate which causes significant economic losses in fish aquaculture. This study investigated the effects of chitosan microspheres cross linked with glutaraldehyde and containing beta-cyclodextrin (betaCD) on the survival of this parasite in 7 d cultures. When used alone in assays, neither chitosan nor betaCD showed any activity, whereas free glutaraldehyde was strongly toxic to the parasite. Microspheres were likewise strongly toxic, at total glutaraldehyde concentrations much lower than with free glutaraldehyde: near-100% ciliate death was obtained (1) with 50 microg ml(-1) of microspheres prepared with 5% glutaraldehyde and no betaCD, or (2) with 10 microg ml(-1) of microspheres prepared with 0.15% glutaraldehyde and 0.1% betaCD. This suggests that the main active component is glutaraldehyde, but that the presence of small amounts of betaCD enhances efficacy. This high efficacy, together with the low toxicity to fish and rapid biodegradability of the individual components, suggest that these microspheres may be an attractive alternative to the formaldehyde baths traditionally used for the control of this parasite.

Analysis of Variance↗

PubMed2005

Effect of cis-resveratrol on genes involved in nuclear factor kappa B signaling.

This study investigated for the first time the effects of the cis isomer of RESV (c-RESV), a polyphenol present in red wine, on an array of genes whose expression is controlled by nuclear factor kappa B (NF-kappaB) and whose transcriptional activation is critical in a number of pathologies (including some cardiovascular diseases). In inflammatory peritoneal macrophages stimulated with lipopolysaccharide (LPS) and gamma interferon (IFN-gamma), c-RESV significantly blocked the expression of genes related to the REL/NF-kappaB/IkappaB family, adhesion molecules and acute-phase proteins; however, the greatest modulatory effect was obtained on the expression of genes related to the pro-inflammatory cytokines. c-RESV down-regulated the nuclear factor of kappa light chain gene enhancer in B-cells 1 (NFkappaBL1) gene product p105 and up-regulated the nuclear factor of kappa light chain gene enhancer in B-cells inhibitor alpha (IkappaBalpha) gene. c-RESV also significantly inhibited intercellular adhesion molecule-1 (ICAM-1) gene expression and the transmembrane receptors RIP (receptor TNFRSF) and TLR3 (toll-like receptor 7). At 100 muM, c-RESV significantly inhibited transcription of Scya2 (chemokine MCP-1), the chemokine RANTES (regulated on activation, normal T cell expressed and secreted), pro-inflammatory cytokines that attract monocyte-granulocyte cells such as M-CSF (colony-stimulating factor 1), GM-CSF (colony-stimulating factor 2) and G-CSF (colony-stimulating factor 3), the cytokine tumor growth factor beta (TGF-beta) and the extracellular ligand IL-1alpha. In contrast, c-RESV stimulated transcription of the pro-inflammatory cytokines IL-6 and tumor necrosis factor alpha (TNF-alpha), the extracellular ligand IL-1beta, and the IFN regulatory factor (IRF)-1. In conclusion, c-RESV has a significant modulatory effect on the NF-kappaB signaling pathway and, consequently, an important antioxidant role that may partially explain the cardioprotective effects attributed to long-term moderate red wine consumption.

Animals↗

PubMed2005

Helminth fauna of the yellow-legged gull Larus cachinnans in Galicia, north-west Spain.

Thirty-six helminth species were found in 324 gulls examined during June 1994 to February 1996 from different localities of Galicia: 25 trematodes (Brachylaima sp., Brachylecithum microtesticulatum, Cardiocephaloides longicollis, Cryptocotyle lingua, Cryptocotyle concavum, Diplostomum spathaceum, Echinostephilla virgula, Galactosomum phalacrocoracis, Gigantobilharzia acotylea, Gymnophallus deliciosus, Gynaecotyla longiintestinata, Himasthla elongata, Himasthla quissetensis, Knipowitschiatrema nicolai, Levinseniella (Levinseniella) propinqua, Maritrema gratiosum, Maritrema linguilla, Microphallus primas, Microphallus similis, Ornithobilharzia canaliculata, Parorchis acanthus, Phagicola minuta, Psilostomum brevicolle, Renicola sp. and Stephanoprora denticulata), four cestodes (Alcataenia micracantha, Microsomacanthus ductilis, Tetrabothrius (Oriana) erostris and Wardium cirrosa), six nematodes (Anisakis simplex, Contracaecum rudolphii, Cosmocephalus obvelatus), Eucoleus contortus, Paracuaria adunca and Tetrameres (Tetrameres) skrjabini) and one acanthocephalan (Arhythmorhynchus longicollis). Tetrabothrius erostris was the most prevalent species (79.6%), followed by C. obvelatus (47.8%), C. lingua (37.4%), G deliciosus (30.9%), G. longiintestinata (22.8%), P. adunca (21.9%), B. microtesticulatum (17.6%), E. contortus (14.5%) and M. similis (9.3%). Microphallus similis was the dominant species, with a Berger-Parker index (BP) of 0.32, followed by T. erostris (BP=0.10). All species presented an aggregated dispersion except G. acotylea and G. phalacrocoracis, which showed a random dispersion. Species that seem to have the greatest predilection for specific sites along the intestine are: C. longicollis and A. micracantha (first third), Brachylaima sp., M. similis and G. longiintestinata (last third) and A. longicollis (second half). Eight species are known to be pathogenic to commercially important fish or molluscan species and several are pathogenic to humans.

Animals↗

PubMed2005

Glugea vincentiae n. sp. (Microsporidia: Glugeidae) infecting the Australian marine fish Vincentia conspersa (Teleostei: Apogonidae).

A parasite of the marine fish Vincentia conspersa was examined by light microscopy and transmission electron microscopy. This parasite develops in the subcutaneous tissue of the body and fins, forming spherical xenomas about 1-2 mm in diameter surrounded by a layer of amorphous material. The observed characteristics of the new parasite are in line with those of the other Glugea species; merogony takes place in the outer zone of the cytoplasm of the host cell, sporogony takes place in sporophorous vesicles, and mature spores are located in the central part of the xenoma. Meronts were cylindrical uninucleate or occasionally triradiate multinucleate, with plasmodia in direct contact with the host cytoplasm. Sporogonic plasmodia divided by multiple cleavage to produce sporoblast mother cells, which after binary fission became sporoblasts. Two types of spores were recognized, both uninucleate, i.e., ovoid or slightly ovoid microspores with a mean size of 5.1 x 2.2 microm and much less frequent as elongated oval macrospores with a mean size of 8.9 x 3.1 microm. The polar tube has between 12 and 14 coils arranged in 1, 2, or 3 layers. Taken together, these characteristics suggest that this microsporidian infecting V. conspersa is a new species of Glugea, which we have named Glugea vincentiae.

Animals↗

PubMed2004

In vitro effects of the polyphenols resveratrol, mangiferin and (-)-epigallocatechin-3-gallate on the scuticociliate fish pathogen Philasterides dicentrarchi.

This study investigated the in vitro effects of the polyphenols resveratrol, mangiferin and (-)-epigallocatechin-3-gallate (EGCG) on the histiophagous ciliate Philasterides dicentrarchi, which causes fatal scuticociliatosis in farmed turbot Scophthalmus maximus L. Of the 3 polyphenols, resveratrol showed strongest antiprotozoal activity, reducing ciliate density after 1 wk culture by, on average, 91% at 50 microM, and 96% at 500 microM. EGCG reduced ciliate density by, on average, 93% at 500 microM, with no significant effect at 50 microM. Mangiferin reduced ciliate density by, on average, 56% at 500 microM, again with no significant effect at 50 microM. In view of these findings, we discuss the potential utility of chemotherapy with polyphenols as a strategy for the control of scuticociliatosis in farmed turbot.

Analysis of Variance↗

PubMed2004

Helminth fauna of Falconiform and Strigiform birds of prey in Galicia, Northwest Spain.

This is a survey of the helminth fauna of 285 individuals of 14 species of birds of prey (Falconiformes and Strigiformes) from Galicia (northwest Spain), namely Buteo buteo, Accipiter nisus, A. gentilis, Milvus migrans, M. milvus, Pernis apivorus, Circus pygargus, Falco tinnunculus, F. peregrinus, F. subbuteo, Tyto alba, Strix aluco, Asio otus and Athene noctua. A total of 15 helminth species were detected, namely 8 nematodes ( Eucoleus dispar, Capillaria tenuissima, Synhimantus laticeps, Microtetrameres sp., Physaloptera alata, Procyrnea leptoptera, Hovorkonema variegatum and Porrocaecum angusticolle), 4 cestodes ( Cladotaenia globifera, Paruterina candelabraria and Mesocestoides sp.), 2 trematodes ( Neodiplostomum attenuatum and Strigea falconis), and 1 acanthocephalan ( Centrorhynchus globocaudatus). The helminth communities observed were basically similar, although there were marked differences in species richness, which was higher in falconiforms (except for A. gentilis) than in strigiforms. More specifically, species richness was highest in B. buteo (13 species), followed by A. nisus (11 species). In the falconiforms, the helminth species present generally exhibited a clear relationship with host diet. In the strigiforms, by contrast, species richness was lower than expected given the host's diet, suggesting that a different explanation is needed.

Animals↗

PubMed2004

Effects of the histiophagous ciliate Philasterides dicentrarchi on turbot phagocyte responses.

Philasterides dicentrarchi is an opportunistic histiophagous ciliate parasite causing systemic scuticociliatosis in cultured turbot (Scophthalmus maximus L.). This study investigated the effects of inoculation with live or killed trophozoites of this ciliate (plus 3% thioglycollate) on the in vitro phagocytic activity and respiratory-burst responses of inflammatory peritoneal leucocytes obtained from the fish thus treated. The phagocytic activity of leucocytes from fish inoculated with killed P. dicentrarchi was higher in the presence than in the absence of infected turbot serum (ITS). The effect of ITS was smaller in fish inoculated with live P. dicentrarchi, indicating modulation of the opsonic activity of ITS. Inoculation with live ciliates led to a significant increase in subsequent in vitro extracellular ROS production, but only when normal turbot serum (NTS) or ITS was included in the assay medium. Inclusion of live P. dicentrarchi in the medium abolished this increase, suggesting ROS-scavenging activity. Inoculation with live P. dicentrarchi led to a significant decline in subsequent in vitro intracellular ROS production; when NTS was included in the medium, there was a significant increase in intracellular ROS production, but no such increase was observed when ITS was included in the medium. Inoculation with live P. dicentrarchi alone did not increase subsequent in vitro NO? production in response to LPS; a significant increase was observed when NTS or ITS was included in the assay medium, but this increase was not affected by prior inoculation with P. dicentrarchi. These results suggest that the amphizoic nature of this parasite may reflect the ease with which it can develop mechanisms of evasion of the host immune response.

Analysis of Variance↗

PubMed2004

An Anacardiaceae preparation reduces the expression of inflammation-related genes in murine macrophages.

This study investigated the effects of an aqueous extract of the stem bark of Mangifera indica L. (Anacardiaceae; Vimang), which contains a defined mixture of components including polyphenols (principally mangiferin, MA), triterpenes, phytosteroids, fatty acids and microelements, on expression of inflammation mediators in inflammatory murine macrophages after stimulation in vitro with lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma). In vitro treatment with Vimang at 4 microg/ml reduced levels of NOS-2 mRNA and NOS-2, while treatment at 40 microg/ml also reduced levels of COX-2 mRNA, COX-2, and prostaglandin E2 (PGE2). Results suggested that MA is involved in these effects. In vitro treatment with Vimang at 40 microg/ml also inhibited mRNA levels of the proinflammatory cytokines interleukin 1beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha) and colony-stimulating factor (GM-CSF), but did not affect mRNA levels of IL-6 or tumor growth factor-beta (TGF-beta). Extracellular release of TNF-alpha by inflammatory macrophages was inhibited by in vitro treatment with Vimang at the same concentrations that showed inhibition of TNF-alpha mRNA levels. The inhibition of TNF-alpha production appears to be at least partially attributable to MA. Vimang at 4 microg/ml decreased mRNA levels of nuclear factor-kappaB (NF-kappaB) but did not affect expression of the NF-kappaB inhibitor (IkappaB). These data indicate that the potent anti-inflammatory effects of Vimang are due to selective modulation of the expression of inflammation-related genes, leading to attenuation of macrophage activation.

Animals↗

PubMed2004

Chemotactic responses of the fish-parasitic scuticociliate Philasterides dicentrarchi to blood and blood components of the turbot Scophthalmus maximus, evaluated using a new microplate multiassay.

This study describes a new capillary-type microplate multiassay for characterization of protozoal chemotactic responses, allowing up to 32 assays to be run simultaneously. We used the new multiassay to evaluate the chemoattractant activity of turbot blood components and turbot cells for the facultative parasite Philasterides dicentrarchi, which is responsible for significant losses in turbot farming. Preliminary tests indicated that the assay requires 3-4 h for detection of chemoattractant activity, that it can be performed effectively using the ciliate axenic culture medium, and that it distinguishes clearly between different concentrations of chemoattractant. Application of the assay indicated that whole blood and serum from normal turbot, and especially infected turbot, have strong chemoattractant activity for P. dicentrarchi trophozoites, whereas neither turbot blood cells nor other turbot cells nor bacteria were significant chemoattractants. These results raise the possibility that turbot serum components are involved in host detection and host invasion by P. dicentrarchi, in line with previous findings indicating that turbot with skin lesions show increased susceptibility to P. dicentrarchi infection.

Animals↗

PubMed2004

Cysteine proteinase activities in the fish pathogen Philasterides dicentrarchi (Ciliophora: Scuticociliatida).

This study investigated protease activities in a crude extract and in vitro excretion/secretion (E/S) products of Philasterides dicentrarchi, a ciliate fish parasite causing economically significant losses in aquaculture. Gelatin/SDS-PAGE analysis (pH 4, reducing conditions) detected 7 bands with gelatinolytic activity (approximate molecular weights 30-63 kDa) in the crude extract. The banding pattern observed in analysis of E/S products was practically identical, except for 1 low-molecular-weight band detected in the crude extract but not in the E/S products. In assays with synthetic peptide p-nitroanilide substrates, the crude extract hydrolysed substrates characteristic of cysteine proteases, namely Z-Arg-Arg pNA, Bz-Phe-Val-Arg pNA and Z-Phe-Arg pNA. These activities were strongly inhibited by the cysteine protease inhibitor E-64 and by Ac-Leu-Val-Lys aldehyde, a potent inhibitor of cysteine proteases of the cathepsin B protease subfamily. The proteases present in the crude extract degraded both type-I collagen and haemoglobin in vitro, consistent with roles in tissue invasion and nutrition respectively. Again, E-64 completely (collagen) or markedly (haemoglobin) inhibited this degradation. Finally, the histolytic activity of the ciliate in turbot fibroblast monolayers was strongly reduced in the presence of E-64, confirming the importance of secreted cysteine proteinases in the biology of Philasterides dicentrarchi.

Animals↗

PubMed2003

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

Philasterides dicentrarchi is a scuticociliate causing fatal disease in farmed turbot and sea bass. In view of its high virulence and endoparasitic location, this parasite cannot be effectively controlled by formalin baths, and no systemic chemotherapeutic treatments have yet proved effective; immunoprophylaxis may thus be an attractive alternative approach. Since vaccine development is greatly facilitated by axenic culture of the pathogen, we have developed a simple axenic culture system based on commercially available Leibovitz L-15 medium, supplemented with fetal bovine serum, lipids (lecithin and Tween 80), nucleosides and glucose. After 1 week's culture under optimal conditions (salinity 10 per thousand, pH 7.2, temperature between 18 and 23 degrees C), yields of 1-2 x 10(5)cells/ml were obtained. Even cultures with seeding densities as low as 20 cells/ml were found to produce a good yield of ciliates (about 6 x 10(4)cells/ml) after 11 days of incubation. The ciliates thus obtained were free of contamination by other microorganisms, enabling preparation of pure P. dicentrarchi antigens for vaccine development studies.

Animals↗

PubMed2003

Mangifera indica L. extract (Vimang) and mangiferin modulate mouse humoral immune responses.

The present study investigated the effects of orally administered Vimang (an aqueous extract of Mangifera indica) and mangiferin (the major polyphenol present in Vimang) on mouse antibody responses induced by inoculation with spores of microsporidian parasites. Inoculation induced specific antibody production with an exponential timecourse, peaking after about one month. Vimang significantly inhibited this antibody production from about three weeks post-inoculation, and most markedly by four weeks post-inoculation; by contrast, mangiferin had no significant effect. Determination of Ig isotypes showed that the IgM to IgG switch began about four weeks post-inoculation, with IgG2a predominating. Vimang significantly inhibited IgG production, but had no effect on IgM. Mangiferin did no affect either IgM or IgG2a, but significantly enhanced production of IgG1 and IgG2b. Neither Vimang nor mangiferin enhanced specific antibody secretion by splenic plasma cells from mice inoculated with microsporidian spores, whether administered in vivo before serum extraction or in vitro to the culture medium. Inoculation with spores induced splenomegaly, which was significantly reduced by Vimang and significantly enhanced by mangiferin. These results suggest that components of Mangifera indica extracts may be of potential value for modulating the humoral response in different immunopathological disorders.

Adjuvants, Immunologic↗

PubMed2003

Anthelminthic and antiallergic activities of Mangifera indica L. stem bark components Vimang and mangiferin.

This study investigated the antiallergic and anthelmintic properties of Vimang (an aqueous extract of Mangifera indica family stem bark) and mangiferin (the major polyphenol present in Vimang) administered orally to mice experimentally infected with the nematode, Trichinella spiralis. Treatment with Vimang or mangiferin (500 or 50 mg per kg body weight per day, respectively) throughout the parasite life cycle led to a significant decline in the number of parasite larvae encysted in the musculature; however, neither treatment was effective against adults in the gut. Treatment with Vimang or mangiferin likewise led to a significant decline in serum levels of specific anti-Trichinella IgE, throughout the parasite life cycle. Finally, oral treatment of rats with Vimang or mangiferin, daily for 50 days, inhibited mast cell degranulation as evaluated by the passive cutaneous anaphylaxis test (sensitization with infected mouse serum with a high IgE titre, then stimulation with the cytosolic fraction of T. spiralis muscle larvae). Since IgE plays a key role in the pathogenesis of allergic diseases, these results suggest that Vimang and mangiferin may be useful in the treatment of diseases of this type.

Administration, Oral↗

PubMed2003

Philasterides dicentrarchi (Ciliophora:Scuticociliatida) expresses surface immobilization antigens that probably induce protective immune responses in turbot.

Philasterides dicentrarchi is a histophagous ciliate causing systemic scuticociliatosis in cultured turbot. This study demonstrates that turbot which survive this disease have serum antibodies that recognize ciliary antigens of this ciliate in ELISA and immobilize/agglutinate the ciliate in vitro. Mouse sera raised against ciliary antigens and integral membrane proteins are likewise capable of immobilizing/agglutinating the ciliates, indicating that P. dicentrarchi, like other ciliates, expresses surface immobilization antigens. Furthermore, the antigen agglutinating reaction induces the parasite to shed its surface antigens rapidly, replacing them with others with different specific serology. This antigen shedding and variation response is similar to that detected in other protozoan parasites. Immunization of turbot with ciliate lysate plus adjuvant or with formalin-fixed ciliates induced synthesis of agglutinating antibodies and conferred a degree of protection against challenge infection, suggesting that the response to surface antigens may play an important role in defence against this pathogen, SDS-PAGE and immunoblotting studies indicated the existence of a predominant polypeptide of about 38 kDa in the ciliary antigen and membrane protein fractions, and this may be the principal surface antigen of P. dicentrarchi.

Agglutination↗

PubMed2002

Histodytes microocellatus gen. et sp. nov. (Dracunculoidea: Guyanemidae), a parasite of Raja microocellata on the European Atlantic coast (north-western Spain).

We describe a new genus, Histodytes, within the family Guyanemidae (Nematoda: Spirurida: Camallanina: Dracunculoidea). The type species, Histodytes microocellatus n. sp., is found in the gill, heart, kidney, spleen and gonad tissues of Raja microocellata from the continental shelf off the estuary of Muros y Noia (north-western Iberian Peninsula). Histodytes differs morphologically from the three other genera described to date in this family ( Guyanema, Travassosnema, Pseudodelphys) because the vulva is situated a long way back from the oesophageal-intestinal union, and the anterior uterine branch almost reaches to the level of this union. In addition, it can be distinguished from Guyanema and Travassosnema by the absence of caudal alae in the male, and from Travassosnema by the much greater length of the glandular oesophagus and the lack of an oesophageal appendix. Histodytes is the only guyanemid genus described to date from an elasmobranch and the first one to be found on the European Atlantic coast.

Animals↗

PubMed2002

Mouse antibody response to a microsporidian parasite following inoculation with a gene coding for parasite ribosomal RNA.

This study found that a plasmid construct encoding the small-subunit ribosomal RNA (SSUrRNA) of the microsporidian Microgemma caulleryi generates a humoral response upon intramuscular inoculation in mice. The plasmid used was pCMV, following preliminary trials indicating efficient beta-galactosidase gene expression in mouse muscle cells transfected with pCMV/beta-Gal. The antibodies produced after inoculation with pCMV/SSUDNA recognized parasite spore antigens and reached maximum levels at 30 days postinoculation, subsequently remaining stable for at least 120 days. Due to the highly conserved sequence of the SSUrDNA in different microsporidian species, these results open up interesting prospects for broad-spectrum vaccination.

Animals↗