RESUMEN
Efficient implementation of an environmental biomarker requires multi-annual comparability over a wide geographical range. The present study improved the comparability of a quantitative competitive metallothionein (MT) enzyme-linked-immuno-sorbent-assay (ELISA) in the sentinel fish Lithognathus mormyrus by introducing to the assay recombinant MT and beta-actin standards. Commercial antibodies for cod MT and mammalian actin were implemented. In addition, a sensitive anti L. mormyrus MT antibody was produced, adequate only for solid phase immunochemical assays. Cadmium was applied to the fish through injection and feeding to serve as a testing platform of the ELISA. The results demonstrated high potential protective capacity of the liver against toxic levels of transition metals through increasing MT levels. MT transcript levels were evaluated also from fish sampled at polluted and relatively clean natural sites, indicating applicability of MT as biomarker of exposure to a multi-factorial pollution, in comparison to its low revealed sensitivity to controlled cadmium exposure.
Asunto(s)
Cadmio/toxicidad , Metalotioneína/análisis , Metalotioneína/inmunología , Perciformes/metabolismo , Contaminantes del Agua/toxicidad , Actinas/genética , Actinas/metabolismo , Animales , Biomarcadores , Cadmio/metabolismo , Ecosistema , Monitoreo del Ambiente , Ensayo de Inmunoadsorción Enzimática/métodos , Hígado/química , Metalotioneína/biosíntesis , Metalotioneína/genética , Perciformes/genética , Proteínas Recombinantes/inmunología , Contaminantes del Agua/metabolismoRESUMEN
A new progressive, fatal disease called 'cup cell disease' was characterized in ex situ cultures of Botryllus schlosseri, a colonial tunicate. The disease originated as a few dark spots growing within zooids. The infected colonies then started to deteriorate, morphologically diagnosed by ampullar retraction, lethargic blood circulation and by a swollen and soft tunic matrix. In later stages of the disease, developed buds were also affected. Many large black dots were scattered within the tunic matrix, and zooids were transformed to opaque, dilated, sac-like structures, signaling impending death. Colonies were infected periodically, even without direct tissue contact. The time course from first appearance to colony death ranged between 30 and 45 d. Histological studies, in vitro culturing of blood cells and blood smears revealed the existence of numerous cup-like cells (up to 4.8 microm diameter on average) with a yellowish cell wall and transparent cytoplasm that was not stained by various dyes (except azocarmine-G). Cells were refractive under bright-field illumination and revealed a flattened wall with flanges, characteristic of species of the phylum Haplosporidia. Cup cells aggregated in blood vessels and in internal parts of zooids and buds and were phagocytosed by blood cells. In a single case, plasmodia-like structures were found only in the tunic matrix of an infected colony. This is the first record in botryllid ascidians of an infectious lethal disease associated with haplosporidian protists.
Asunto(s)
Haplosporidios , Urocordados/parasitología , Animales , Acuicultura , California , Técnicas HistológicasRESUMEN
Colonies of the urochordate Botryllus schlosseri may fuse upon contact if they share common alleles on the highly polymorphic fusibility/histocompatibility locus. While, in these chimeras, one of the partners is usually morphologically eliminated (resorbed), circulating totipotent cells of the inferior genotype on the resorption phenomenon may parasitize either the soma or the germ line of the winner. Here, we show an environmental split of the two stem cell lineages that may develop germ cell parasitism vs somatic cell cooperation. Each naturally formed Botryllus chimera can be a composite of component genotypes created through two unlinked parasitic germ and somatic cell lineage interactions. The germ line parasitism is inherited through a pedigree. Conversely, by using amplified fragment length polymorphism (AFLP) and microsatellite alleles as polymorphic genetic markers, and seawater temperature as the variable environmental factor, we documented that the somatic constituent of chimeric zooids was shifted from one genotype to another, in accordance with the changes in seawater temperatures. This variable somatic state of chimerism in the field may, thus, carry benefits to the chimeral entity, which presents synergistically, at any time, the best-fitted combination of its genetic components.