178
M.R. Coscia and U. Oreste
Once obtained, the complete nucleotide and deduced amino acid
sequences will help understand the structural characteristics accounting for
the adaptation of the Antarctic fish Ig to low temperatures.
Humoral Immune Response of Antarctic Fish Against
Nematode Parasites
The nematode species most frequently found infesting fish are Anisakis
simplex [10], Contracaecum osculatum [11] and Pseudoterranova
decipiens [12], all belonging to Anisakidae family. Fish are intermediate
hosts of the third stage nematode larvae and marine mammals are the final
hosts of the adult parasites; the liver is the main site of infestation [13]. The
nematode proteins involved in fish immune response certainly include
secreted/excreted protein and the components of the cuticle (collagens,
cuticlins and surface-associated proteins)[14].
Fish immune response against parasites has been poorly investigated: the
presence of Anguilla anguilla antibodies, specific for the nematode
Anguillicola crass us [15], the monogenean Pseudodactylogyrus bini [16]
and the trematode Telogaster opisthorchis [17] has been demonstrated.
We addressed our interests to the molecular bases of the parasitism of
Antarctic fish by nematodes: our first aim was to verify the presence of
Antarctic fish antibodies specific for nematode antigens; a further goal was
the purification of both antibodies and nematode antigens.
Nematodes were collected from the liver of specimens of different
teleostean species: those recovered from N coriiceps, C. aceratus, T.
bernacchii, Chionodraco rastrospinosus, Pseudochaenichthys georgian us
and Cryodraco antarcticus were identified as third stage larvae of
Pseudoterranova decipiens, whereas those from C. hamatus as third stage
larvae of Contracaecum osculatum complex. Both the nematode species
belong to the superfamily of Ascaridoidea, family Anisakidae, subfamily
Anisakinae.
Some P. decipiens nematodes were individually maintained alive in
vessels containing phosphate buffer saline (PBS) and the proteins released
within 8 days were collected: this preparation, containing the
secreted/excreted proteins, was referred to as PdRP. Dead larvae were
incubated, at 37 °e, in PBS containing a cocktail of protease inhibitors; the
extract obtained (PdAP) contained, presumably, the cuticular components
corresponding to surface-associated proteins. Finally, the larvae were
boiled for 10 min in the lysis buffer containing 2% SDS and 10% f3mercaptoethanol and incubated in the same buffer, at 37 °e overnight. This
preparation (PdSP) presumably contained collagens and cuticlins in
addition to solubilized cellular components. The extract of C. osculatum
M.R. Coscia and U. Oreste
Once obtained, the complete nucleotide and deduced amino acid
sequences will help understand the structural characteristics accounting for
the adaptation of the Antarctic fish Ig to low temperatures.
Humoral Immune Response of Antarctic Fish Against
Nematode Parasites
The nematode species most frequently found infesting fish are Anisakis
simplex [10], Contracaecum osculatum [11] and Pseudoterranova
decipiens [12], all belonging to Anisakidae family. Fish are intermediate
hosts of the third stage nematode larvae and marine mammals are the final
hosts of the adult parasites; the liver is the main site of infestation [13]. The
nematode proteins involved in fish immune response certainly include
secreted/excreted protein and the components of the cuticle (collagens,
cuticlins and surface-associated proteins)[14].
Fish immune response against parasites has been poorly investigated: the
presence of Anguilla anguilla antibodies, specific for the nematode
Anguillicola crass us [15], the monogenean Pseudodactylogyrus bini [16]
and the trematode Telogaster opisthorchis [17] has been demonstrated.
We addressed our interests to the molecular bases of the parasitism of
Antarctic fish by nematodes: our first aim was to verify the presence of
Antarctic fish antibodies specific for nematode antigens; a further goal was
the purification of both antibodies and nematode antigens.
Nematodes were collected from the liver of specimens of different
teleostean species: those recovered from N coriiceps, C. aceratus, T.
bernacchii, Chionodraco rastrospinosus, Pseudochaenichthys georgian us
and Cryodraco antarcticus were identified as third stage larvae of
Pseudoterranova decipiens, whereas those from C. hamatus as third stage
larvae of Contracaecum osculatum complex. Both the nematode species
belong to the superfamily of Ascaridoidea, family Anisakidae, subfamily
Anisakinae.
Some P. decipiens nematodes were individually maintained alive in
vessels containing phosphate buffer saline (PBS) and the proteins released
within 8 days were collected: this preparation, containing the
secreted/excreted proteins, was referred to as PdRP. Dead larvae were
incubated, at 37 °e, in PBS containing a cocktail of protease inhibitors; the
extract obtained (PdAP) contained, presumably, the cuticular components
corresponding to surface-associated proteins. Finally, the larvae were
boiled for 10 min in the lysis buffer containing 2% SDS and 10% f3mercaptoethanol and incubated in the same buffer, at 37 °e overnight. This
preparation (PdSP) presumably contained collagens and cuticlins in
addition to solubilized cellular components. The extract of C. osculatum