福氏志贺菌mdoC基因对其在洗菜水中生长及生物膜形成的影响

【目的】研究福氏志贺菌mdoC基因对细菌在不同洗菜水中生长及生物膜形成的影响。【方法】以福氏志贺菌野生型和敲除mdoC基因的opgC突变体为出发菌株,采用生长曲线法和结晶紫染色法,在低渗透压及正常渗透压的菠菜、芹菜、生菜及白菜洗菜水中,研究福氏志贺菌野生型和opgC突变体生长及生物膜的产生能力。【结果】在不同洗菜水中,福氏志贺菌opgC突变体生长速率明显较野生型慢,加盐提高渗透压之后,野生型和opgC突变体稳定生长期均提前。野生型和opgC突变体的生物膜产量在菠菜水、生菜水和白菜水中有显著区别;提高渗透压之后,福氏志贺菌野生型在菠菜水中的生物膜产量显著提高,而在生菜水和白菜水中生物膜产量显著下降,opgC突变体在菠菜水、生菜水和白菜水中生物膜产量均显著提高。【结论】低渗条件下,福氏志贺菌mdoC基因的缺失显著地延缓了细菌生长。提高渗透压后,在菠菜、生菜和白菜洗菜水中,mdoC基因的缺失促进了生物膜的形成。     

Application of dietary supplements (synbiotics and probiotics in combination with plant products and β‐glucans) in aquaculture

This review addresses the dietary supplements, synbiotics and probiotics in combination with plant products or yeast/β‐glucans in aquaculture. The potential applications of synbiotics have a relatively short history in aquaculture, but have generated interest because of numerous benefits reported in endothermic animals. Since the first study was published by (Aquaculture Science in 2009) the concept has been used in aquaculture to reveal effects on growth performance, gut microbiota, gut histology, immune parameters, haematological and biochemical parameters as well as increased disease resistance. However, a limited number of probiotic bacteria (mainly Bacillus and Enterococcus) and prebiotics (mannan oligosaccharides, fructooligosaccharides, short‐chain fructooligosaccharides, galactooligosaccharides, arabinoxylan–oligosaccharides, isomaltooligosaccharide, chitosan oligosaccharide and inulin) have been used. Additionally, some studies have used plant products or yeast/β‐glucans in combination with probiotics, and these studies suggested that these dietary supplements promote growth performance and boost some immune parameters. The present review identifies evaluations of gut microbiota and gut morphology, and mucosal immune response as significant gaps in existing knowledge and suggests issues that merit further investigations to conclude the potential of dietary supplements in aquaculture.     

Effects of dietary β‐(1,3)(1,6)‐D‐glucan supplementation on growth performance,intestinal morphology and haemato‐immunological profile of mirror carp (Cyprinus carpio L.)

In recent years, aquaculture research has focused on probiotics, prebiotics, and β‐glucans, in order to improve health status and growth performance. Information regarding the effects of β‐glucan on growth performance and intestinal immunity of mirror carp (Cyprinus carpio L.) is scarce. An experiment was therefore conducted to investigate the effects of a yeast β‐glucan preparation (MacroGard^®) on growth performance, intestinal morphology and haemato‐immunological indices of mirror carp. Carp (initial weight 11.1 ± 0.0 g) were fed highly purified diets supplemented with 0% (control), 0.1%, 1% or 2% MacroGard^® for 8 weeks. Fish fed diets containing 1% and 2% MacroGard^® showed significant improvements in weight gain, specific growth rate and feed conversion ratio compared to fish fed both the control and the 0.1% MacroGard^® containing diet. Histological appraisal of the intestine showed a significantly higher infiltration of leucocytes into the epithelial layer of fish fed diets supplemented with 1% and 2% MacroGard^® in the anterior intestine compared to fish fed the control and 0.1% MacroGard^® diet. This effect was not observed in the posterior intestine. There were no significant differences in the intestinal absorptive surface area and number of goblet cells in either intestinal region. At the end of the experiment, the haematological status of the fish was examined. Compared to control fed fish, the haematocrit value was significantly elevated in fish fed the 2% MacroGard^® diet. Furthermore, the blood monocyte fraction was significantly higher in fish fed the 1% and 2% MacroGard^® diets. No significant changes were observed in the other blood parameters assessed. The present study shows that high dietary β‐glucan inclusion increases growth performance without detrimental effects on the health indicators assessed. Increased intraepithelial leucocytes in the anterior intestine may indicate a localized immune response; no detrimental effects on intestinal morphology were observed.     

Effect of treatment of chum salmon Oncorhynchus keta (Walbaum) eggs with 1,3;1,6‐β‐D‐glucans on their development and susceptibility to Saprolegnia infection

The effects of six 1,3;1,6‐β‐D‐glucooligo‐ and polysaccharides with different structures (ranging from 1 to 10 kDa in molecular mass and containing 10–25% of β‐1,6‐linked glucose residues) from brown algae, Saccharina cichorioides, on development of the chum salmon, Oncorhynchus keta (Walbaum), were evaluated. Exposure of chum salmon eggs to 1,3;1,6‐β‐D‐glucans with a molecular mass of more than 2 kDa increased the survival of embryos and juveniles and their resistance to Saprolegnia infection by up to 2.5‐fold, leading to a weight gain in juveniles of 40–55% compared with The control chum salmons. The 1,3;1,6‐β‐D‐glucans with molecular mass of 6–8 kDa and used at a at concentration of 0.5 mg mL^?1 rendered the best stimulative effect.     

The effect of heat treatment on the soluble protein content of oats

The effect of heat treatment on the soluble protein content in oat groats (Kerstin commercial variety) was evaluated using asymmetric flow field-flow fractionation (AF4) in combination with online multiangle light scattering (MALS) and UV detection. The AF4 method was used to separate the monomeric proteins from globulin hexamer and aggregate proteins and β-glucan polysaccharides in the soluble oat protein fraction. The total amount of soluble protein (with respect to total protein) was reduced to 35.7 ± 4.5 wt. % in heat treated oats from 74.6 ± 5.3 wt. % in non-heat treated oats. The ratio of monomeric to globulin hexamer and aggregate proteins was reduced from 1.82 to 1.48 as a result of heat treatment. Sodium-dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed the selective elimination of protein bands associated with the albumin and prolamin protein fractions as a result of heat treatment. These results were supported through amino acid analysis by cation exchange chromatography coupled with UV detection which revealed a reduction in amino acid residues associated with prolamin. The globulin proteins were found to be less sensitive to heat treatment.     

Biochemical parameters in the blood of gilthead sea bream (Sparus aurata Linnaeus, 1758) supplemented with commercially available β‐glucan‐based product (IMUNO‐2865®)

The aim of this study was to evaluate the effects of a promising new immune stimulant in aquaculture (IMUNO‐2865^®) on biochemical parameters in sea bream during the winter stress period. A total of 640 sea bream were fed throughout 90 days with diets containing 0 (Group 1), 1 (Group 2), 10 (Group 3) and 25 g (Group 4) of IMUNO‐2865^® kg^?1 of feed. Samples were taken each month and 90 days after supplementation. No statistical differences among treatment groups were noticed for the following biochemical parameters: glucose (GLU), aspartate aminotransferase (AST), plasma cholesterol (CHOL), lactate dehydrogenase (LDH), urea (URE) and creatinine (CREA). At the final sampling, total ammonia (NH₃) was higher in Groups 3 and 4 compared to the control and the low supplementation group (p < .05), while total proteins (TP) was significantly higher in Group 4 compared to all other groups, and in Groups 2 and 3 compared to the control group (p < .05). Blood Ca⁺⁺ levels were significantly higher after 60 days of feeding in all treatment groups compared to the control, and remained elevated in Group 4 even after 90 days following cessation of supplementation (p < .05). The results of this study described the increase of biochemical parameters in the blood of sea bream after use of IMUNO‐2865^® but future research is needed to evaluate its potentially immunostimulative effect on fish in commercial aquaculture.     

Dietary β‐glucan modulate haematological parameters,cytokines and gene expression in TLR and ERK pathways of rainbow trout (Oncorhynchus mykiss) during infection by Aeromonas salmonicida

To explore the role of β‐glucan (0, 0.05%, 0.1% and 0.2%) in resisting bacteria, rainbow trout (Oncorhynchus mykiss) was challenged with Aeromonas salmonicida after 42‐day β‐glucan administration, and then sampled at 0, 2, 4 and 6 days post infection (dpi). The data showed that 0.2% β‐glucan reduced the accumulated mortality rates compared with the ICG (infected control group) (p < .05). The white blood cells, red blood cells and haemoglobin were higher in the 0.2% β‐glucan group (BG) than the ICG (p < .05). 0.1% and 0.2% β‐glucan elevated serum total antioxidative capability and glutathione activity but alleviated the increase of serum alkaline phosphatase activity and glucose concentration (p < .05) during infection. Serum TNF‐α, IL‐1β, IL‐8 and IgM in three BGs elevated remarkably on 6 dpi compared with the ICG (p < .05). Expression of tnf, il1b and cxcl8 of the head kidney in the 0.1% and 0.2% BGs was higher than the ICG on 4 dpi while ighm expression in the 0.2% BG was higher than in the ICG on 2 and 6 dpi (p < .05). 0.1% and 0.2% β‐glucan increased the expression of tlr5m, tlr5s, tmek and myd88 in the spleen after infection (p < .05). Similarly, 0.2% β‐glucan up‐regulated the expression of tmek, myd88, oncmyk‐dab and c3 in head kidney (p < .05). Overall, 0.2% dietary β‐glucan effectively decreased accumulated mortality rate by modulating the biochemistry process, cytokines, and activating TLR and ERK signalling pathways during A. salmonicida infection.     

糖原组学研究进展——β-葡聚糖的结构和功能的研究进展

糖原组学是研究糖类的结构及其功能特性特别是糖类发挥作用的生物学机制的一门新型学科，将成为生命科学研究中的新热点。多糖及其缀合物在生命活动过程中贮存着各种生物信息，在细胞内外之间传递各种物质。目前，β-葡聚糖的研究是该领域研究最为广泛的糖类，其中研究最早的是香菇多糖，因为它具有免疫活性和抗肿瘤作用；作者对多糖的生物学地位以及β-葡聚糖的结构及其与生物学功能的关系的研究进展作一简述。葡聚糖的生物活性与其分子量、溶解度、黏度、支化度、链构象及结晶度有关，β-葡聚糖具有多种不同的构象，一重和三重螺旋是其存在的主要高级结构形式。     

1,3‐1‐6 ß‐glucans enhance tissue regeneration in zebrafish (Danio rerio): Potential advantages for aquaculture applications

High aquacultural rearing density and handling of fish may frequently result in skin or gills wounds, thereby facilitating the onset of secondary infections. The capacity of the zebrafish to regenerate tissues, as well as fins and other organs, makes it an ideal animal model for studying the mechanisms of tissue regeneration. Since macrophages are involved in tissue regeneration, a diet including ß‐glucans might positively affect the process through activation of macrophages and other immune pathways. Consequently, the aim of this study was to investigate the effects of the oral administration of 1,3‐1,6 β‐glucans on the regeneration process of the caudal fin after its amputation in zebrafish. One hundred and twenty zebrafish were randomly distributed into four groups with three replicates each: an untreated non‐amputated group (CNA) and an untreated amputated group (CA) fed a control diet; two treated and amputated groups (MI and MII) fed for 14 days the same diet with the addition of two differently extracted 1,3‐1,6 ß‐glucans (MacroGard^® and Experimental MacroGard^®, Biorigin^©). ß‐glucans were added to allowed the administration of 12.5 mg/kg of fish body weight (0.35 g/kg of feed). Results showed that 1,3‐1,6 ß‐glucans decreased fish mortality rate and enhanced both daily and cumulative regenerated fin area, independent of the specific ß‐glucan extraction method used. Based on the mechanisms similarities of the innate immune system and tissue regeneration among different teleost species, these results may likely be extended to species of interest for the aquaculture sector.     

Response of the intestinal mucosal barrier of carp (Cyprinus carpio) to a bacterial challenge by Aeromonas hydrophila intubation after feeding with β‐1,3/1,6‐glucan

The effect of dietary β‐glucan on the bacterial community in the gut of common carp (Cyprinus carpio) was examined after oral application of Aeromonas hydrophila. Carp received either feed supplemented with 1% MacroGard^®, a β‐1,3/1,6‐glucan, or a β‐glucan‐free diet. Fourteen days after feeding, half of the carp from each group were intubated with 10⁹ colony‐forming units (CFU) of a pathogenic strain of A. hydrophila. Gut samples were taken 12 hr to 7 days after application and analysed using microbiological and molecular biological techniques (NGS, RT‐PCR‐DGGE). The reaction of the mucosa and the microbiota to an A. hydrophila intubation differed in carp fed with β‐glucan compared to carp from the control group. In β‐glucan fed carp, the total bacterial amount was lower but the number of bacterial species was higher. Bacterial composition was different for carp from both treatment groups. The number of mucin filled goblet cells was reduced in carp fed the β‐glucan diet. Mucus was obviously released from the goblet cells and was probably washed out of the gut together with high numbers of bacteria. This might be protective against pathogenic bacteria and, therefore, feeding with β‐glucan may provide protection against infections of the gut in carp.