The objective of this study was to evaluate dietary supplementation with different copper sulphate (CuSO ) forms on small intestine microanatomy and large intestine microbiota. Ninety weaned piglets were divided into three experimental groups: control diet (CTR), with no added CuSO and diets supplemented with 150 ppm of CuSO in protected (150P) and unprotected form (150UP). After 18 days of dietary treatment, six piglets per treatment were randomly selected and sacrificed. Duodenum villi length and crypt depths were higher (P < 0.001) in the animals fed 150UP than other groups. Glial fibrillary acidic protein (GFAP), a marker for enteric glial cells, was unaffected by dietary treatments. The total bacteria and Enterobacteriaceae bacteria counts were lower (P < 0.05) in cecum of animals fed 150P in comparison with the other two groups. In the colon the Streptococci spp were lower (P < 0.001) in both CuSO supplemented groups than controls. The obtained results revealed a modulation of intestinal structure and microbiota exerted by the studied CuSO dietary supplementation. The present data show that dietary supplementation with 150UP in the first period post-weaning may assist in restoring the gut morphology, improving duodenal structure.
Environmental temperature is one of the critical factors affecting fish development. The aim of this study was to examine the impact of three different rearing temperatures (16, 19 and 22°C) throughout the endogenous feeding phase of the Siberian sturgeon Acipenser baerii. This was performed by assessing (a) larval survival and growth; (b) immunofluorescence localization and expression of genes involved in muscle development and growth – myog and Igf1; and (c) stress status through the expression of thermal stress genes – Hsp70, Hsp90α and Hsp90β – and whole body cortisol. Overall survival rate and larval weight did not differ significantly across temperatures. Larvae subjected to 22°C showed faster absorption of the yolk‐sac than larvae subjected to 19 or 16°C. Both at schooling and at the end of the trial, larvae reared at 16°C showed significantly lower levels of cortisol than those reared at 19 or 22°C. IGF‐1 immunopositivity was particularly evident in red muscle at schooling stage in all temperatures. The expression of all Hsps as well as the myog and Igf1 genes was statistically higher in larvae reared at 16°C but limited to the schooling stage. Cortisol levels were higher in larvae at 22°C, probably because of the higher metabolism demand rather than a stress response. The observed apparent incongruity between Hsps gene expression and cortisol levels could be due to the lack of a mature system. Further studies are necessary, especially regarding the exogenous feeding phase, in order to better understand if this species is actually sensitive to thermal stress.
This study evaluated the effects of rearing density on muscle growth and development in Siberian sturgeon (Acipenser baerii) larvae. Three different stocking densities were tested: low (LD, 30 larvae/l), mid (MD, 80 larvae/l) and high (HD, 150 larvae/l) in a recirculating aquaculture system. Larvae were sampled at hatching (T0), schooling (T1) and complete yolk-sac absorption (T2) stage and were weighed and processed for muscle tissue histometrical analyses and for qualitative morphological study analyses; fatty acid profile was also determined by Gas Chromatography-Flame Ionization Detector analysis. Low-density larvae presented a higher weight than MD or HD at T2 (p < 0.05). Histometrical analysis revealed that total muscle area was similar at T1 and T2, but higher than T0, while it was lower at HD at schooling (p < 0.05). The fatty acid profile revealed no differences between densities while, during development, there was a selective consumption: sparing or increasing of essential fatty acids to the detriment of their precursors. Our study suggests that lower densities appear to be more suitable to rear Siberian sturgeon in this particular stage of development. Indeed, larvae reared at the lower density were heavier and longer while larvae reared at the higher density showed lower muscle proliferation rate. As a consequence, LD larvae may exert an increase of potential growth at a mid-long term. K E Y W O R D S density, fatty acids, larvae, muscle structure, Siberian sturgeon 1 | INTRODUCTION During the past two centuries, the natural stocks of Siberian sturgeon (Acipenser baerii) suffered a sharp decline, due to overfishing, pollution and loss of spawning spots. In 1998, all sturgeon species were effectively added to the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES), which allowed to control the illegal trade of sturgeons and their products and the implementation of conservation plans.Survival and growth during early stages of development of Siberian sturgeon and throughout the following life periods is of great importance both for conservation aquaculture production programs and for commercial purposes. Following an increased demand of commercial production facilities, there is a growing need for the improvement of hatchery technologies that allow the production of high quality Siberian sturgeon larvae. After hatching, only endogenous feeding occurs (Balon, 2001), when larvae entirely rely on the yolk-sac reserves for energy and growth, until its digestive system is
Menisci play an essential role in shock absorption, joint stability, load resistance and its transmission thanks to their conformation. Adult menisci can be divided in three zones based on the vascularization: an avascular inner zone with no blood supply, a fully vascularized outer zone, and an intermediate zone. This organization, in addition to the incomplete knowledge about meniscal biology, composition, and gene expression, makes meniscal regeneration still one of the major challenges both in orthopedics and in tissue engineering. To overcome this issue, we aimed to investigate the role of hypoxia in the differentiation of the three anatomical areas of newborn piglet menisci (anterior horn (A), central body (C), and posterior horn (P)) and its effects on vascular factors. After sample collection, menisci were divided in A, C, P, and they were cultured in vitro under hypoxic (1% O2) and normoxic (21% O2) conditions at four different experimental time points (T0 = day of explant; T7 = day 7; T10 = day 10; T14 = day 14); samples were then evaluated through immune, histological, and molecular analyses, cell morpho-functional characteristics; with particular focus on matrix composition and expression of vascular factors. It was observed that hypoxia retained the initial phenotype of cells and induced extracellular matrix production resembling a mature tissue. Hypoxia also modulated the expression of angiogenic factors, especially in the early phase of the study. Thus, we observed that hypoxia contributes to the fibro-chondrogenic differentiation with the involvement of angiogenic factors, especially in the posterior horn, which corresponds to the predominant weight-bearing portion.
The success of cell-based approaches for the treatment of cartilage or fibro-cartilaginous tissue defects requires an optimal cell source with chondrogenic differentiation ability that maintains its differentiated properties and stability following implantation. For this purpose, the aim of this study was to evaluate the use of endostatin (COL18A1), an anti-angiogenic factor, which is physiologically involved in cell differentiation during meniscus development. Swine neonatal meniscal cells not yet subjected to mechanical stimuli were extracted, cultured in fibrin hydrogel scaffolds, and treated at two different time points (T1 = 9 days and T2 = 21 days) with different concentrations of COL18A1 (10 ng/mL; 100 ng/mL; 200 ng/mL). At the end of the treatments, the scaffolds were examined through biochemical, molecular, and histochemical analyses. The results showed that the higher concentration of COL18A1 promotes a fibro-chondrogenic phenotype and improves cellularity index (DNA content, p < 0.001) and cell efficiency (GAGs/DNA ratio, p < 0.01) after 21 days. These data are supported by the molecular analysis of collagen type I (COL1A1, a marker of fibrous-like tissue, p < 0.001), collagen type II (COL2A1, a marker of cartilaginous-like tissue, p < 0.001) and SRY-Box Transcription Factor 9 (SOX9, an early marker of chondrogenicity, p < 0.001), as well as by histological analysis (Safranin-O staining), laying the foundations for future studies evaluating the involvement of 3D endostatin hydrogel scaffolds in the differentiation of avascular tissues.
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