Identifying and Characterizing a Novel Peritrophic Matrix Protein (MdPM-17) Associated With Antibacterial Response From the Housefly,Musca domestica (Diptera: Muscidae)

Peritrophic matrix/membrane (PM) critically prevents the midgut of insects from external invasion by microbes. The proteins in the peritrophic membrane are its major structural components. Additionally, they determine the formation and function of this membrane. However, the role of PM proteins in immune regulation is unclear. Herein, we isolated a novel PM protein (MdPM-17) from Musca domestica larvae. Further, the function of MdPM-17 in regulating host innate immunity was identified. Results showed that the cDNA of MdPM-17 full is 635 bp in length. Moreover, it consists of a 477-bp open reading frame encoding 158 amino acid residues. These amino acid residues are composed of two Chitin-binding type-2 domain (ChtBD2) and 19 amino acids as a signal peptide. Moreover, tissue distribution analysis indicates that MdPM-17 was enriched expressed in midgut, and moderate levels in the fat body, foregut, and malpighian tubule. Notably, MdPM-17 recombinant protein showed high chitin-binding capacity, thus belongs to the Class III PM protein group. MdPM-17 protein silencing via RNA interference resulted in the expression of antimicrobial peptide (defensin, cecropins, and diptericin) genes, and this occurred after oral inoculation with exogenous microbes Escherichia coli (Enterobacteriales:Enterobacteriaceae), Staphylococcus aureus (Bacillales:Staphylococcaceae), and Candida albicans (Endomycetales:Saccharomycetaceae)). Therefore, all the antimicrobial peptide (AMP) gene expression levels are high in MdPM-17-depleted larvae during microbial infection compared to controls. Consequently, these findings indicate that MdPM-17 protein is associated with the antibacterial response from the housefly.     

Marine-Derived Chitosan Nanoparticles Improved the Intestinal Histo-Morphometrical Features in Association with the Health and Immune Response of Grey Mullet (Liza ramada)

Marine-derived substances are known for their beneficial influences on aquatic animals’ performances and are recommended to improve intestinal health, immunity, and anti-oxidative status. The present study investigates the role of chitosan nanoparticles on the intestinal histo-morphometrical features in association with the health and immune response of Grey Mullet (Liza ramada). Chitosan nanoparticles are included in the diets at 0, 0.5, 1, and 2 g/kg and introduced to fish in a successive feeding trial for eight weeks. The final body weight (FBW), weight gain (WG), and specific growth rate (SGR) parameters are significantly increased while feed conversion ratio (FCR) decreases by chitosan nanoparticles compared to the control (p < 0.05). The morphometric analysis of the intestines reveals a significant improvement in villus height, villus width, and the number of goblet cells in chitosan-treated groups in a dose-dependent manner. Additionally, there is a positive correlation between the thickness of the enterocyte brush border and the chitosan dose, referring to an increasing absorptive activity. Histologically, the intestinal wall of Grey Mullet consists of four layers; mucosa, sub-mucosa, tunica muscularis (muscular layers), and serosa. The histological examination of the L. ramada intestine shows a normal histo-morphology. The epithelial layer of intestinal mucosa is thrown into elongated finger-like projections, the intestinal villi. The values of hemoglobin, hematocrit, red blood cells (RBCs), total protein (TP), albumin, and globulin are significantly increased in fish fed 1, and 2 g/kg of chitosan nanoparticles compared to fish fed 0 and 0.5 g/kg (p < 0.05). The highest levels of TP and albumin are observed in fish fed 1 g/kg diet (p < 0.05). The lysozyme activity and phagocytic index are significantly enhanced by feeding chitosan nanoparticles at 0.5, 1, and 2 g/kg, whereas the phagocytic activity is improved in fish fed 1 and 2 g/kg (p < 0.05). The highest lysozyme activity and phagocytic index are observed in fish fed 1 g/kg. SOD is significantly activated by feeding chitosan nanoparticles at 1 g/kg. Simultaneously, glutathione peroxidase (GPx) and catalase (CAT) activities also are enhanced by feeding chitosan at 1 and 2 g/kg, compared to fish fed 0 and 0.5 g/kg (p < 0.05). The highest GPx and CAT activities are observed in fish fed 1 g/kg (p < 0.05). Conversely, the malondialdehyde (MDA) levels are decreased by feeding chitosan at 1 and 2 g/kg, with the lowest being in fish fed 1 g/kg (p < 0.05). To summarize, the results elucidate that L. ramada fed dietary chitosan nanoparticles have a marked growth rate, immune response, and anti-oxidative response. These improvements are attributed to the potential role of chitosan nanoparticles in enhancing intestinal histo-morphometry and intestinal health. These results soundly support the possibility of using chitosan nanoparticles at 1–2 g/kg as a feasible functional supplement for aquatic animals.     

Evaluation of Intestinal Absorption of Dietary Halocynthiaxanthin,a Carotenoid from the Sea Squirt Halocynthia roretzi

Halocynthiaxanthin is an acetylenic carotenoid mainly found in Halocynthia roretzi. To date, several bioactivities of halocynthiaxanthin have been reported, but its mechanism of digestion and absorption in mammals has not been studied yet. In this study, we evaluated the intestinal absorption of halocynthiaxanthin in mice. The halocynthiaxanthin-rich fraction was prepared from the tunicate Halocynthia roretzi. Mice were orally administered the fraction at a dose of 5 mg/kg body weight. The halocynthiaxanthin levels in the plasma, liver, and small intestine, were quantified using HPLC-PDA, 1, 3, 6, and 9 h after ingestion. The halocynthiaxanthin-rich fraction mainly consisted of the all-trans form and a small amount of cis forms. These three isomers were detected in the plasma of mice 3 h after ingestion. Time-course changes after the ingestion of this fraction were found, with cis isomers being more abundant than the all-trans isomer in the mouse plasma and liver. In the small intestine, however, the all-trans isomer was primarily detected. The possibility that cis isomers might be absorbed rapidly from the small intestine cannot be denied, but our results suggest that dietary all-trans-halocynthiaxanthin might be isomerized to the cis isomer after intestinal absorption.     

重庆地区草地贪夜蛾幼虫肠道真菌的分离鉴定

草地贪夜蛾(Spodoptera frugiperda)是一种洲际迁飞性农业害虫,食谱广泛. 2019年5月已迁飞至重庆地区,对农作物造成严重危害.为了解入侵重庆地区草地贪夜蛾(Spodoptera frugiperda)肠道真菌菌群的组成,本研究以采集自重庆巫山玉米田及重庆江津高粱田中的草地贪夜蛾幼虫为材料,运用传统培养方法分离其粪便中的肠道优势真菌,并对其进行形态学观察和分子生物学鉴定.实验通过分离培养不同地区、不同食物的草地贪夜蛾肠道真菌,结合形态学观察及rDNA ITS测序完成了属水平的鉴定,共分离得到了5个属10个真菌分离株,其中分离自重庆巫山玉米田的草地贪夜蛾肠道真菌归为3个属,分别为念珠菌属(Candida)、掷孢酵母属(Sporobolomyces)和帚枝霉属(Sarocladium);分离自重庆江津高粱田的草地贪夜蛾肠道真菌归为3个属,分别为念珠菌属(Candida)、莫氏黑粉菌属(Moesziomyces)和毛霉属(Mucor).初步分离获得入侵重庆地区的草地贪夜蛾肠道真菌,丰富了对入侵草地贪夜蛾肠道微生物的认识.     

重庆地区取食高粱的草地贪夜蛾肠道细菌新分离株的鉴定

为充分认识入侵重庆地区草地贪夜蛾(Spodoptera frugiperda)的肠道微生物, 2019年7月该课题组再次在重庆江津地区高粱地采集了草地贪夜蛾幼虫,通过分离培养结合16S rDNA测序鉴定,从江津草地贪夜蛾幼虫肠道中共分离得到38个细菌分离株,归为9个属,分别为克雷伯氏菌属(Klebsiella)、肠球菌属(Enterococcus)、微球菌属(Micrococcus)、葡萄球菌属(Staphylococcus)、芽孢杆菌属(Bacillus)、不动杆菌属(Acinetobacter)、寡养单胞菌属(Stenotrophomonas)、假单胞菌属(Pseudomonas)和鞘脂杆菌属(Sphingobacterium).在课题组前期研究的基础上,新分离到了6个属,分别为克雷伯氏菌属(Klebsiella)、微球菌属(Micrococcus)、葡萄球菌属(Staphylococcus)、芽孢杆菌属(Bacillus)、寡养单胞菌属(Stenotrophomonas)和鞘脂杆菌属(Sphingobacterium).同时还对入侵重庆地区采食高粱和玉米的草地贪夜蛾幼虫肠道细菌分离株进行了比较分析,只在采食高粱的草地贪夜蛾幼虫肠道中发现了欧文氏菌属(Erwinia)、微球菌属(Micrococcus)、葡萄球菌属(Staphylococcus)、芽孢杆菌属(Bacillus)和寡养单胞菌属(Stenotrophomonas)的菌株,而气单胞菌属(Aeromonas)、短波单胞菌属(Brevundimonas)、金黄杆菌属(Chryseobacterium)和类香味菌属(Myroides)则只在采食玉米的草地贪夜蛾幼虫肠道中分离得到.通过对同一片江津高粱地的草地贪夜蛾和玉米黏虫(Mythimna separata)的肠道细菌分离株进行比较分析,只在草地贪夜蛾肠道中发现了微球菌属(Micrococcus)、欧文氏菌属(Erwinia)、假单胞菌属(Pseudomonas)、葡萄球菌属(Staphylococcus)和鞘脂杆菌属(Sphingobacterium),而金黄杆菌属(Chryseobacterium)只发现于玉米黏虫肠道中.     

北京鸭中脑脊髓通路的起源和细胞构筑：HRP逆行追踪法研究

在脊髓的颈中部、颈膨大部和腰膨大部单侧注射或包埋HRP(辣根过氧化物酶),逆行追踪了28例北京鸭,对中脑至脊髓的传导通路的起始部位、细胞构筑和机能进行了系统的研究。发现大量标记细胞分布在中脑对侧的红核,双测的Cajal中介核、中央灰质和动眼神经副交感核(即EW核)内。此外,还发现少量标记细胞分布在中脑中缝核和双侧的中脑外侧网状结构。而在顶盖内,没有任何部位出现标记细胞。研究结果表明,北京鸭除具有红核脊髓束外,还存在着Cajal中介核至脊髓的直接传导通路,EW核至脊髓的直接传导通路以及中缝核和网状结构至脊髓的直接传导通路,而不存在与哺乳类相似的顶盖脊髓束。     

植物源乳酸菌对断奶仔猪日粮表观消化率及消化道内容物特性的影响

试验利用植物源乳酸菌,对断奶仔猪日粮中养分表观消化率及消化道内容物特性的影响进行了研究。结果表明,仔猪日粮中添加植物源乳酸菌可分别提高干物质、粗蛋白、能量、粗脂肪、钙、磷等的表观消化率;降低消化道pH,增加肠道乳酸菌的数量,可弥补仔猪胃酸分泌不足。同时,还可减少消化道盲肠和结肠内大肠杆菌的数量,改善动物消化道微生态平衡。     

壳聚糖饲用微生物制剂对湘黄鸡免疫指标和肠道菌群的影响

在不用药物、不接种疫苗的条件下,于饮水中添加0（CK）,0．5％,1．0％和1．5％的以壳聚糖为培养底物研制的壳聚糖饲用微生物制剂饲喂湘黄鸡,探讨其对湘黄鸡免疫指标和肠道菌群的影响效果．结果表明,试验组脾脏指数有上升趋势（Jp〈0．25）,法氏囊指数和胸腺指数显著提高（p〈0．05）;试验组血液中IgG含量呈上升趋势（P〈0．25）,T3,T4和IgA含量也显著提高（P〈0．05或（p〈0．01）;试验组盲肠中大肠杆菌显著减少,而乳酸杆菌显著增加（P〈0．01）．同时在日增重和育成率上,试验组提高率达9．31％（p〈0．01）和7．01％（p〈O．05）,表明该制剂通过促进免疫器官发育,增强T3,T4,IgA,IgG分泌和改善肠道微生态环境,从而提高试鸡的抗病力和生长性能．     

罗非鱼消化道5-HT内分泌细胞的免疫组织化学研究

用链霉亲和素-生物素化过氧化物酶复合物(Strept Avidin Biotion -peroxidase Complex SABC )免疫细胞化学方法,对罗非鱼消化道内分泌细胞进行了免疫组织化学定位.结果表明:5-羟色胺细胞在消化道各段都有分布,其中胃体部密度最大,其次是幽门和喷门处,后肠的密度最小.本文主要讨论了5-HT内分泌细胞在罗非鱼消化道免疫细胞化学定位,可为鱼类的内分泌学提供主要依据.     

甘露寡糖对肉鸡生产性能和肠道微生物以及免疫机能的影响

为探明甘露寡糖在肉鸡生产上的最佳使用剂量,选用11日龄广东岭南黄肉鸡1200只,随机分为6组,每组4个重复,每重复50只鸡,分别饲喂6种不同日粮,自由采食和饮水.第1组为空白对照组,第2,3,4,5,6组为试验组,在基础日粮的基础上分别添加1g/kgBio-MOS(甘露寡糖),4mg/kg黄霉素,1g/kgBio-MOS+4mg/kg黄霉素,0.5g/kgBio-MOS和1.5g/kgBio-MOS.结果表明:(1)添加Bio-MOS,提高了肉鸡的日增重和采食量,但与对照组之间差异不显著(p>0.05).(2)添加Bio-MOS1g/kg,盲肠中双歧杆菌显著高于对照组(p<0.05),但对回肠中双歧杆菌增殖效果不明显.与对照组相比,添加不同水平的Bio-MOS对盲、回肠乳酸杆菌增殖效果不明显,但不同程度地提高了盲肠中乳酸杆菌的含量;盲肠中大肠杆菌数量没有明显降低;回肠中除0.5g/kg组外,其余各组之间大肠杆菌含量差异不显著.(3)与对照组相比,添加Bio-MOS1g/kg极显著地提高了血液中T淋巴细胞的百分率(p<0.01).添加Bio-MOS对血清中新城疫抗体效价影响不明显,但以饲料中添加Bio-MOS1g/kg效果最好.饲料中添加Bio-MOS对血清中IgA,IgG,IgM影响不显著,但添加Bio-MOS1g/kg,其IgA,IgG,IgM的含量明显高于其他各组.(4)肉鸡日粮中同时添加甘露寡糖和黄霉素,两者表现出协同作用,但不及甘露寡糖单独使用时的效果.