RFamide peptides are expressed in the early stages of development in most vertebrates. Gonadotrophin‐inhibitory hormone (GnIH) belongs to the RFamide family, and its role in reproduction has been widely studied in adult vertebrates, ranging from fish to mammals. Because only 3 reports have evaluated GnIH during development, the present study aimed to characterise the ontogeny of GnIH in the fish model Cichlasoma dimerus. We detected the presence of 2 GnIH‐immunoreactive (‐IR) cell clusters with spatial and temporal differences. One cluster was observed by 3 days post‐hatching (dph) in the nucleus olfacto‐retinalis (NOR) and the other in the nucleus posterioris periventricularis by 14 dph. The number of GnIH‐IR neurones increased in both nuclei, whereas their size increased only in the NOR from hatchling to juvenile stages. These changes occurred from the moment that larvae started feeding exogenously and during development and differentiation of gonadal primordia. We showed by double‐label immunofluorescence that only GnIH‐IR neurones in the NOR co‐expressed gonadotrophin‐releasing hormone 3 associated peptide. In addition, GnIH‐IR fibre density increased in all brain regions from 5 dph. GnIH‐IR fibres were also detected in the retina, optic tract and optic tectum, suggesting that GnIH acts as a neuromodulator of photoreception and the integration of different sensory modalities. Furthermore, there were GnIH‐IR fibres in the pituitary from 14 dph, which were in close association with somatotrophs. Moreover, GnIH‐IR fibres were observed in the saccus vasculosus from 30 dph, suggesting a potential role of GnIH in the modulation of its function. Finally, we found that gnih was expressed from 1 dph, and that the pattern of variation of its transcript levels was in accordance with that of cell number. The results of the present study comprise a starting point for the study of new GnIH roles during development.

Vitamin K (VK) is a fat-soluble vitamin that vertebrates have to acquire from the diet, since they are not able to de novo synthesize it. VK has been historically known to be required for the control of blood coagulation, and more recently, bone development and homeostasis. Our understanding of the VK metabolism and the VK-related molecular pathways has been also increased, and the two main VK-related pathways—the pregnane X receptor (PXR) transactivation and the co-factor role on the γ-glutamyl carboxylation of the VK dependent proteins—have been thoroughly investigated during the last decades. Although several studies evidenced how VK may have a broader VK biological function than previously thought, including the reproduction, little is known about the specific molecular pathways. In vertebrates, sex differentiation and gametogenesis are tightly regulated processes through a highly complex molecular, cellular and tissue crosstalk. Here, VK metabolism and related pathways, as well as how gametogenesis might be impacted by VK nutritional status, will be reviewed. Critical knowledge gaps and future perspectives on how the different VK-related pathways come into play on vertebrate’s reproduction will be identified and proposed. The present review will pave the research progress to warrant a successful reproductive status through VK nutritional interventions as well as towards the establishment of reliable biomarkers for determining proper nutritional VK status in vertebrates.

Aquaculture’s sustainability deeply relies on the identification and inclusion of alternative raw materials. Although meals from insects and/or byproducts from different industries are being recently tested, the meal from terrestrial vegetable species is still the main substitution candidate for fish meal. Here the effects of 0% (Control), 10% (A10) and 30% (A30) inclusion of Narbonne vetch (Vicia narbonensis; ZV-156 strain) meal in rainbow trout (Oncorhynchus mykiss) diets was assessed in a 63-day feeding trial by means of growth performance, histopathological, nutritional value of the fish fillet and blood biochemistry analyses. A dose-response trial was conducted in triplicate with 25 rainbow trout juveniles (20 g average body weight) per 500 L tank. Narbonne vetch meal decreased total intestine protease activity in vitro (from 26.81% to 48% inhibition), although high temperature partially inhibited the action of antinutritional factors (ANFs). No differences in fish growth performance and no severe histopathological alterations on the proximal intestine were observed between 10% Narbonne vetch inclusion and Control groups. In contrast, high inclusion (30%) of Narbonne vetch led to poor growth performance (30% reduction on final growth) and severe histopathological alterations (e.g., loss of brush border integrity, high number of villi fusion, reduced goblet cells density as well as reduced width of submucosa, muscular and serosa layers). Furthermore, while the A30 diet decreased docosahexaenoic fatty acid (FA) content in fish fillets, the A10 diet improved monounsaturated FA content when compared to that of the Control group. No altered levels of cholesterol, glucose or triglycerides in blood plasma and/or histopathological effects on the liver were observed among fish fed the different experimental diets. Although further research efforts (e.g., identifying potential enzymatic treatments to decrease the action of ANFs from Narbonne vetch meal) might be required, present results show that a low inclusion (10%) of Narbonne vetch in rainbow trout diets is possible. The inclusion of locally produced legumes such a Narbonne vetch might be an interesting approach to reduce carbon footprint in European aquaculture and the dependency on other alternative raw materials such as soybean (Glycine max) imported from third countries.

Skeletal deformities reduce fish viability, growth, wellbeing, and feed efficiency but also degrade the consumer’s perception of aquaculture products. Herein, the skeletal development and the incidence of skeletal deformities in tench (Tinca tinca) reared in semi-extensive conditions has been described in detail for the first time. Larval skeletons were assessed through an acid-free double-staining procedure in 157 individuals, while 274 specimens at the juvenile stage were evaluated through X-ray analysis. The first skeletal structures to be formed were those related with breathing and feeding activities (e.g., Meckel’s cartilage and opercula) and were visible in larvae of 4 mm of standard length (SL). The axial skeleton was fully ossified in larvae of 12–17 mm of SL, and the caudal fin complex in larvae with 17–26 mm of SL. At the larval stage, no upper-jaw or opercula deformities were observed, while a low incidence (1–9%) of other severe deformities in the heads of the fish (e.g., lower-jaw deformities) were reported. The incidence of vertebral deformities in tench reared in natural ponds was considerable in larvae (54%) and juveniles (52%). Vertebral deformities (fusion and compression) were the most common deformities found in tench larvae (approximately 30%) and vertebral shape deformity in juveniles (around 10%), being mainly located in the caudal region. Thus, a regular monitoring of the skeletal deformities in tench might help to identify better rearing protocols and improve product quality sold at markets. Characterizing the skeletal development not only in semi-extensive systems such as artificial and natural ponds but also under intensive rearing conditions, seems vital for a sustainable and profitable European tench aquaculture.