ABSTRACT
AIMS: To produce and characterize egg yolk immunoglobulin (IgY) against the fish intracellular pathogen Piscirickettsia salmonis as well as to evaluate the antibacterial activity of IgY in vitro and the availability in the serum of fish immunized orally. METHODS AND RESULTS: Specific IgY was produced by immunizing hens with P. salmonis proteins. The IgY was obtained from egg yolks using the ammonium sulphate precipitation method and it was characterized by SDS-PAGE, Western-blot and ELISA, demonstrating that anti-P. salmonis IgY strongly reacted specifically against P. salmonis proteins. In an in vitro neutralization assay, IgY inhibited the growth of P. salmonis in liquid medium at concentrations ranging from 128 to 256 µg ml(-1) in a dose-dependent manner. Interestingly, IgY against P. salmonis also generates a strong protective effect on the infection of P. salmonis in salmon head kidney-1 cells. In addition, the bacteriostatic function of IgY appears to result possibly from agglutination by the interaction of IgY with surface components of the pathogen. Finally, to confirm this IgY as an alternative for salmonid treatment, Atlantic salmon (Salmo salar) specimens were orally inoculated with IgY. The analysis of the sera demonstrates that IgY was effectively transported by fish intestine and that this immunoglobulins maintains its properties and recognizes several proteins of P. salmonis up to 12 h after inoculation of IgY against P. salmonis. CONCLUSIONS: Specific IgY effectively inhibited the growth of P. salmonis and this immunoglobulin can be released in the Atlantic salmon sera when administered orally to fish. SIGNIFICANCE AND IMPACT OF THE STUDY: We propose that this specific IgY against this fastidious micro-organism could be a useful strategy for the treatment of piscirickettsiosis.
Subject(s)
Anti-Bacterial Agents/pharmacology , Egg Yolk/chemistry , Fish Diseases/microbiology , Immunoglobulins/pharmacology , Piscirickettsia/drug effects , Piscirickettsiaceae Infections/veterinary , Animals , Anti-Bacterial Agents/isolation & purification , Chickens/immunology , Electrophoresis, Polyacrylamide Gel , Fish Diseases/drug therapy , Fish Diseases/immunology , Immunoglobulins/isolation & purification , Piscirickettsia/growth & development , Piscirickettsiaceae Infections/drug therapy , Piscirickettsiaceae Infections/immunology , Piscirickettsiaceae Infections/microbiology , Salmo salar/microbiologyABSTRACT
The appearance, persistence and eventual decline of IgG, IgM and IgA antibodies recognizing Cryptosporidium parvum antigens were studied in naturally infected lambs using a Western blot technique, and the results compared with those obtained using sera from immunized lambs. There was an intense recognition of some low molecular weight proteins (15-17 kDa by IgG and IgA; 28-30 kDa by IgA, IgM and IgG) during the infection and early post-infection period. These peptides were not recognized after Days 45-60 of life. Some high molecular weight antigens (94 kDa) were weakly recognized on Day 15 but more intensely recognized from Day 30 onwards, persisting until at least Day 90. Antibody recognition of these C. parvum proteins could be an indicator of recent or past exposure to the parasite.
Subject(s)
Antibodies, Protozoan/immunology , Antigens, Protozoan/immunology , Cryptosporidiosis/immunology , Cryptosporidium parvum/immunology , Sheep Diseases/immunology , Animals , Antigens, Protozoan/chemistry , Blotting, Western , Immune Sera/immunology , Immunization/veterinary , Immunoglobulins/immunology , Molecular Weight , SheepABSTRACT
The immunoglobulin (IgG), IgM and IgA responses in Castellana-Manchega cross-bred colostrum-deprived and colostrum-fed lambs infected neonatally with Cryptosporidium parvum were measured using an enzyme-linked immunosorbent assay. A comparison of oocyst shedding and anti-C. parvum serum IgG levels in lambs suffering either natural or experimental infection was undertaken. Effects on the oocyst shedding and IgG levels of C. parvum rechallenge at 30 and 120 days of age in neonatally infected lambs were also evaluated. Anti-C. parvum immunoglobulin levels in colostrum-deprived animals peaked on Day 30 of life for IgG and on Day 15 for IgM and IgA. Lambs that received maternal colostrum showed elevated anti-C. parvum IgG, IgM and IgA levels at 3 days old indicating a transfer of colostral immunoglobulins. Experimentally infected lambs showed a IgG response similar to naturally infected lambs, suggesting that the serum IgG response is independent of the infective dose. Finally, lambs rechallenged at 30 and 120 days old did not show either appreciable oocyst shedding or any increase in their anti-C. parvum IgG levels when compared with prechallenged animals.
Subject(s)
Antibodies, Protozoan/blood , Cryptosporidiosis/immunology , Cryptosporidium parvum/immunology , Immunoglobulins/blood , Sheep Diseases/immunology , Animals , Colostrum/immunology , Diarrhea/immunology , Diarrhea/veterinary , Feces/parasitology , Immunity, Maternally-Acquired , Immunoglobulin A/blood , Immunoglobulin G/blood , Immunoglobulin M/blood , SheepABSTRACT
Polyclonal antibodies raised against Cryptosporidium parvum oocysts were found to cross-react with Eimeria spp. oocyst antigens in an indirect immunofluorescence assay, and sera from Eimeria spp.-infected lambs reacted with some antigens from sonicated C. parvum oocysts (between 29 to 30 and 66 to 69 kDa) by Western blot (immunoblot). No cross-reaction was observed with cystozoites of Toxoplasma and Sarcocystis spp. These results show the existence of epitopes common to C. parvum and various Eimeria spp.
