姜黄素吸收代谢及其对家禽繁殖调控作用研究进展

姜黄素是从姜科植物（如姜黄）、天南星科植物根茎中提取出来的多酚类化合物,主要在动物肠道中进行吸收代谢,具有抗炎、抗氧化、抗凋亡、抗衰老、神经保护、免疫调节及代谢调控等作用。姜黄素在动物体内生物利用度差,为提高姜黄素的利用度,通常将其制成多种制剂及配型（如膜制剂、纤维制剂、乳制剂、水凝胶制剂、胡椒碱、半萜类化合物、环糊精、葫芦巴膳食纤维和卵磷脂）。不同的制剂及配型可增加姜黄素产物稳定性,保障姜黄素的有效利用。家禽主要通过下丘脑 - 性腺轴负反馈调节机制来调控生殖细胞发育,实现繁殖调控。此外,家禽的繁殖性能易受外部因素影响,其中光照的影响最大。光照刺激产生的褪黑素具有强抗氧化性,能通过缓解卵巢和睾丸组织的线粒体功能障碍来抑制凋亡和衰老,保障下丘脑 - 性腺轴负反馈调节机制的稳定和氧化应激状态下的卵泡发育。肠道微生物产生的血清素是褪黑素的前体物质,可以影响褪黑素的生成;其次肠道微生物刺激肠上皮细胞中的传入神经直接影响下丘脑分泌促性腺激素释放激素;最后,肠道内产生的雌二醇（E2）可以通过血液进入卵巢组织,并促进卵巢发育。研究发现,姜黄素可通过肠道微生物产生的短链脂肪酸在体内发挥功能作用,但姜黄素是否通过肠道微生物产生的短链脂肪酸来影响家禽的繁殖性能亟待探究。该文综述了姜黄素的生物活性、代谢方式、产品研发利用以及对家禽繁殖调控的作用机制及相关研究进展,旨在为姜黄素作为饲料添加剂在家禽生产中的合理应用提供理论参考。     

低温胁迫诱导斑马鱼ZF4细胞ROS及MAPK相关蛋白表达的影响

为了研究鱼类低温下分子调控机制及其与活性氧(reactive oxygen species,ROS)之间的关系,本实验对斑马鱼(Danio rerio)胚胎成纤维ZF4细胞进行不同程度的低温胁迫(18℃和10℃),监测其在不同低温胁迫时间下(1 d、3 d和5 d)ROS的变化以及丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)通路中蛋白的表达情况。结果显示:(1)DCFH-DA探针法表明在低温胁迫作用下,细胞内ROS含量增加。细胞内ROS上升水平与外界胁迫压力成正相关。低温处理3 d后,18℃和10℃的细胞,其ROS含量与对照组(28℃)相比分别显著升高到(1.23±0.04)倍(P0.05)和(2.31±0.08)倍(P0.05)。(2)Western Blot检测磷酸化的p38(p-p38)和磷酸化的JNK(p-JNK p54和p-JNK p46)的水平变化,结果显示低温胁迫可以使p38和JNK的活性增强,且均在10℃处理3 d达到最高水平。(3)进一步检测DNA断裂标记蛋白γH2A.X的表达水平,结果显示,无论在18℃还是10℃,其在第3天呈现高表达。本实验初步证实低温胁迫能够诱导斑马鱼细胞ROS的产生;通过对不同时间点的蛋白表达水平检测,发现p-JNK、p-p38和γH2A.X激活时间点与低温诱导ROS表达时间点相吻合。该研究为后期对斑马鱼细胞低温胁迫实验奠定基础,其中低温下处理3 d可以作为一个检测各个蛋白变化的关键时间点。     

Fluorescence localization of epidermal cathepsins L and B in the Japanese eel

Histochemical localization of proteolytic activities in the dorsal epidermis of Japanese eel was demonstrated by fluorescent microscopy utilizing 4-methoxy-2-naphthylamide (4M$\beta$NA) derivatives as substrates and 5-nitrosalicylaldehyde as a trapping agent. Carbobenzoxy-L-phenylalanyl-L-arginyl-4M$\beta$NA (Cbz-Phe-Arg-4M$\beta$NA) and Cbz-Arg-Arg-4M$\beta$NA were used for direct detection of cathepsins L and B activities, respectively, in fresh frozen sections and unfixed cells of the eel epidermis. The fluorescing areas, where Cbz-Phe-Arg-4M$\beta$NA was hydrolyzed by cathepsin L, were shown in mucus secretory cells and club cells and broadly around skin surface. The fluorescing areas due to Cbz-Arg-Arg-4M$\beta$NA hydrolysis by cathepsin B were localized similarly in these tissues. The fluorescing intensity for both catheptic activities in mucus secretory cells was higher than that in club cells, where small fluorescing granules were distributed. These results indicate that eel cathepsins L and B are stored in epidermal secretory cells at different levels and probably serve as defense factors before or after secretion by these cells. Abbreviations: Cbz – carbobenzoxy; 4M$\beta$NA – 4-methoxy-2-naphthylamide; NSA – 5-nitrosalicylaldehyde.     

底播虾夷扇贝活品供应链各环节中肠道菌群分析

以鲜活底播虾夷扇贝为研究对象,研究扇贝肠道菌群结构与其生长环境的相似性,并跟踪研究组成供应链的采捕、活水运输、中转、净化Ⅰ、净化Ⅱ及市场等6个环节的活品扇贝的肠道菌群结构变化规律。首先,在扇贝捕捞船现场采集扇贝、海水以及海底沉积物,随后继续对活水运输、中转、净化Ⅰ、净化II及市场等环节跟踪采集扇贝肠道样品;利用基于16S rDNA的PCR-变性梯度凝胶电泳指纹图谱技术对扇贝肠道菌群、沉积物及海水进行分析。变性梯度凝胶电泳指纹图谱分析结明,从即捕扇贝肠道、沉积物及海水中分别获得12、18及20个扩增条带,其中,三者具有9条相同的条带;其余包括活水运输、中转、净化Ⅰ、净化Ⅱ及市场等5个环节的条带数依次为14、16、13、9、22。相似性分析表明,即捕扇贝肠道与海水及沉积物之间菌群组成相似性系数（戴斯系数）分别为66．6％和62．5％;由采捕到市场6环节之间虾夷扇贝肠道菌群相似性系数依次为64．0％、73．3％、69．0％、63．6％及32．3％。采捕后至净化池以及净化后至市场均发生菌群条带的增加,表明采捕后流通环境的改变对扇贝肠道菌群结构有影响;净化处理具有有效的减菌效果;现有捕后处置方式即从采捕至净化Ⅱ,活品扇贝处于比较稳定的胁迫状态,而净化后的产品至消费市场环节的胁迫压力明显,即该零售终端环节对活品质量有重要影响。     

Effects of dietary thiamin supplement on growth,body composition and intestinal enzyme activities of juvenile Jian carp (Cyprinus carpio var. Jian)

A total of 1050 juvenile Jian carp (Cyprinus carpio var. Jian) (8.20 ± 0.02 g) were fed diets containing seven graded levels of thiamin (0.25, 0.48, 0.79, 1.06, 1.37, 1.63 and 2.65 mg kg^?1) for 60 days to investigate the effects of thiamin on growth, body composition and digestive enzyme activities. Percent weight gain (PWG), feed intake and feed efficiency (FE) were the lowest in fish fed the basal diet (P < 0.05). Protein productive value and lipid productive value increased with increasing dietary thiamin levels up to 1.06 and 0.79 mg kg^?1 diet, respectively (P < 0.05). Body protein and lipid increased with increasing dietary thiamin levels (P < 0.05), while moisture and ash of fish carcasses decreased with the increase in dietary thiamin supplementation (P < 0.05). Intestinal folds height had a similar trend to PWG (P < 0.05). Activities of α‐amylase, lipase, trypsin, Na⁺, K⁺‐ATPase, alkaline phosphatase and gamma‐glutamyl transpeptidase in intestine were all affected by the dietary thiamin (P < 0.05). In conclusion, thiamin could improve growth and intestinal enzyme activities of juvenile Jian carp. The dietary thiamin requirement of juvenile Jian carp (8.0–60.2 g) based on PWG was 1.02 mg kg^?1 diet.     

Maturation of the pancreatic and intestinal digestive functions in sea bass (Dicentrarchus labrax): effect of weaning with different protein sources

The maturation of the digestive functions in sea bass (Dicentrarchus labrax) larvae was evaluated by the enzymatic profile of pancreas and intestine brush border membranes. Sea bass larvae were weaned at day 25 with three simplified diets different by their protein nature: 100% fish meal (FP), 100% casein mixture (CP) and 50% fish meal-50% casein mixture (CFP). The casein mixture contained 35% of hydrolysate. The control group was fed live preys. The specific activity of amylase decreased with age irrespectively of the diets whereas the specific activity of trypsin was enhanced. The casein mixture reduced pancreatic secretion in amylase and trypsin. The CFP group differed from the other groups fed on compound diets, exhibiting as soon as day 32 high activities of brush border enzymes, similar to controls. This sharp increase between day 25 and 32 appeared to be crucial for larval survival. The addition of a protein hydrolysate in a weaning diet seems to facilitate this maturation process.     

Artificially induced tetraploid masu salmon have the ability to form primordial germ cells

Diploid gametes generated with tetraploid animals are a stepping stone to improving chromosome manipulation techniques. However, artificially induced tetraploid individuals generally die soon after hatching. Diploid gametes could be induced by in vivo cultures of tetraploid primordial germ cells (PGCs) through germ-line chimera. In the present study, characteristics of PGCs were studied in inviable tetraploid masu salmon, Oncorhynchus masou. Histological observation of tetraploid embryos revealed that the same or smaller numbers of PGCs were observed and they migrate into the genital ridges as did diploid PGCs during gonadogenesis. By whole-mount in situ hybridization using vasa messenger RNA (mRNA), 4–35 vasa-positive signals were detected in a pair of genital ridges of tetraploids. By cytological observation of genital ridge cell suspensions, several large round cells were observed, some of which extended pseudopodia. They also contained large nuclei and round granules in their cytoplasm, characteristics of PGCs. As the results suggest that inviable artificial tetraploids have PGCs, we expect to achieve diploid gamete production through surrogate propagation and tetraploid fish production.     

Effects of Cu2+-exchanged montmorillonite on intestinal microflora, digestibility and digestive enzyme activities of Nile tilapia

A total of 360 Nile tilapia at an average initial body weight of 3.9 g were randomly allocated to four treatments, each of which had three replicates of 30 fish per tank and used to investigate the effects of Cu²⁺‐exchanged montmorillonite (Cu‐MMT) on intestinal microflora, digestibility and digestive enzyme activities. The dietary treatments were: (i) basal diet, (ii) basal diet + 1.5 g kg^?1 MMT; (iii) basal diet + 30 mg kg^?1 copper as CuSO₄ (equivalent to the copper in the Cu‐MMT treatment group), or (iv) basal diet + 1.5 g kg^?1 Cu‐MMT. The results showed that supplementation with Cu‐MMT significantly improved growth performance as compared with the control and fish fed with Cu‐MMT had higher growth performance than those fed with MMT or CuSO₄. Supplementation with Cu‐MMT reduced (P < 0.05) the total intestinal aerobic bacterial counts and affected the composition of intestinal microflora with Aeromonas, Vibrio, Pseudomonas, Flavobacterium, Acinetobacter, Alcaligence, Enterobacteriaceae decreasing as compared with the control. Addition of MMT improved (P < 0.05) the apparent digestibility of dry matter and crude protein, and the activities of intestinal amylase, lipase, and alkaline phosphatase as compared with the control. Addition of Cu‐MMT improved (P < 0.05) the apparent digestibilities of dry matter, crude protein, ether extract, and crude ash, and the activities of total protease, amylase, lipase, and alkaline phosphatase as compared with the control. While supplementation with CuSO₄ had no (P > 0.05) effect on intestinal microflora, digestibility, and digestive enzyme activities the results showed that Cu‐MMT exhibited antibacterial activity in vivo resulting in a positive effect on digestive enzyme activities, and then promoted the digestion and absorption of dietary nutrition.     

Impacts of diet on hindgut microbiota and short‐chain fatty acids in grass carp (Ctenopharyngodon idellus)

Diet is known to influence intestinal microbiota in fish, but the specifics of these impacts are still poorly understood. Different protein/fibre ratio diets may result in differing structures and activities of gut microbiota. We examined the hindgut microbiome of grass carp (Ctenopharyngodon idellus) fed three different diets: fish meal (FM, high protein – low fibre), Sudan grass (SG, high fibre – low protein) and compound feed (CF, intermediate). Microbial profiles of fish fed on FM were significantly different from profiles of fish fed CF and SG (F = 18.85, p < .01). Cetobacterium, known to be positively associated with protein digestion, was the dominant microbial group in FM samples (approximately 75.7%), while Lachnospiraceae and Erysipelotrichaceae, thought to be involved in fermentation of plant polysaccharides, were dominant in CF and SG samples (46.8% and 42.9% respectively). Network analyses indicated that the abundance of Lachnospiraceae and Erysipelotrichaceae was in a significantly positive correlation (r = .895, p = .001). Short‐chain fatty acid (SCFA) levels may indicate that the digestibility of diet by microbiota in the grass carp gut decreased from FM to SG (FM>CF>SG). Overall low SCFA levels indicate that hindgut fermentation probably provides a low proportion of energy requirements in grass carp.     

Culture of mantle epithelial cells expressing shell matrix proteins from scallop Patinopecten yessoensis

ABSTRACT:   In molluscs, mantle epithelial cells secrete organic matrix proteins to form shells. In this study, we established a culture of mantle epithelial cells by using the mantle pallial layer of scallops. We aimed to identify the mantle epithelial cells expressing scallop shell matrix proteins and establish a culture system of epithelial cells. After the mantle pallial layer was carefully isolated from the mantle tissue, explant culture was performed at 4°C. Most cells that migrated from the explant tissue were round cells. Most of the adhered cells retained round morphology, while some of the cells adhered to the dish and showed morphology similar to that of epithelial-like and fibroblast-like cells. When the cultured cells were immunostained with a polyclonal antibody against the shell matrix protein, the antibody recognized many of the adhered cells. An estimation of the number of epithelial cells revealed that approximately 70% of the adhered cells were epithelial cells. This is the first report to describe epithelial cells in cultured mantle cells, which express shell matrix proteins. This culture system may be a useful method for characterization of the mantle epithelial cells.