RESUMO
European eel (Anguilla anguilla) is a commercially important species for fisheries and aquaculture in Europe and the attempt to close the lifecycle in captivity is still at pioneering stage. The first feeding stage of this species is characterized by a critical period between 20 to 24 days post hatch (dph), which is associated with mortalities, indicating the point of no return. We hypothesized that this critical period might also be associated with larvae-bacterial interactions and the larval immune status. To test this, bacterial community composition and expression of immune and stress-related genes of hatchery-produced larvae were explored from the end of endogenous feeding (9 dph) until 28 dph, in response to three experimental first-feeding diets (Diet 1, Diet 2 and Diet 3). Changes in the water bacterial community composition were also followed. Results revealed that the larval stress/repair mechanism was activated during this critical period, marked by an upregulated expression of the hsp90 gene, independent of the diet fed. At the same time, a shift towards a potentially detrimental larval bacterial community was observed in all dietary groups. Here, a significant reduction in evenness of the larval bacterial community was observed, and several amplicon sequence variants belonging to potentially harmful bacterial genera were more abundant. This indicates that detrimental larvae-bacteria interactions were likely involved in the mortality observed. Beyond the critical period, the highest survival was registered for larvae fed Diet 3. Interestingly, genes encoding for pathogen recognition receptor TLR18 and complement component C1QC were upregulated in this group, potentially indicating a higher immunocompetency that facilitated a more successful handling of the harmful bacteria that dominated the bacterial community of larvae on 22 dph, ultimately leading to better survival, compared to the other two groups.
Assuntos
Anguilla , Animais , Anguilla/genética , Larva/genética , Dieta/veterinária , Aquicultura , Expressão GênicaRESUMO
Closing the life cycle of European eel (Anguilla anguilla) in captivity is targeted to provide a sustainable, year-round supply of juveniles for aquaculture. Present focus is on the nutritional requirements during the larval first-feeding period. In this study, three experimental diets were tested on hatchery-produced European eel larvae from the onset of the first-feeding stage commencing 10 days post hatch (dph) until 28 dph. Larval mortality was recorded daily, while sampling was conducted at regular intervals to record larval biometrics and analyze the expression of genes related to digestion, appetite, feed intake and growth. Two periods of high mortality were identified: the first appeared shortly after introduction of feeds (10-12 dph), while the second occurred 20-24 dph, indicating the "point of no return". This interpretation was supported at the molecular level by the expression of the gene encoding the "hunger hormone" ghrelin (ghrl) that peaked at 22 dph in all dietary trials, suggesting that most larvae were fasting. However, in larvae fed diet 3, ghrl expression was downregulated beyond 22 dph, which indicated that those larvae were no longer starving at this stage, while upregulation of genes encoding the major digestive enzymes (try, tgl, and amyl2a) advocated their healthy development. Moreover, for larvae fed diet 3, the expression of those genes as well as genes for feed intake (pomca) and growth (gh) continued to increase towards 28 dph. These results together with the registered highest survival, largest dry weight increase, and enhanced biometrics (length and body area) pointed to diet 3 as the best-performing. As a whole, this first-feeding study represents a landmark being the first to document European eel larval growth and survival beyond the point of no return, providing novel insights into the molecular development of digestive functions during the first feeding stage.
