自然微生物挂膜处理水产养殖废水的效果及微生物群落分析

以弹性填料和流化床填料为硝化反应的生物挂膜材料, 聚羟基丁酸/戊酸共聚酯(PHBV)为反硝化反应的碳源和生物膜载体, 通过微生物自然挂膜处理低C/N比水产养殖废水, 去除水体中的氨氮、亚硝酸盐氮及总氮。应用Miseq高通量测序技术对生物膜的微生物群落组成和结构进行分析。结果表明: 温度25—30℃, 该处理系统首次挂膜成功需要4周, 启动后运行稳定, 对2种不同来源和氮污染程度的养殖废水均有较好的脱氮效果, 氨氮、亚硝酸盐氮及总氮的去除率均在90%以上。硝化生物膜(a)的优势菌分别归属变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes)。反硝化生物膜(b)微生物群落的多样性指数和丰度指数均远大于前者, 主要为变形菌门、厚壁菌门、拟杆菌门、螺旋体门(Spirochaetae)及绿菌门(Chlorobi)。其中, 归属于变形菌门β-变形菌纲(Betaproteobacteria)的丛毛单胞菌科(Comamonadaceae)和红环菌科(Rhodocyclaceae)在2种生物膜中占比均较高。由于所处环境(载体, 碳源、溶氧等)不同, 在属分类水平上, 2种生物膜的细菌群落结构表现出明显差异。生物膜a中属的种类仅为b的三分之二, 相对丰度>0.5%的优势菌属, a为8个, b为18个。其中, 隶属丛毛单胞菌科和红环菌科未知属的优势种群分别占到a、b总序列数的56.67%和45.51%。磁螺菌属(Magnetospirillum)和硝化螺菌属(Nitrospira)是a中特有的优势功能菌群, 梭菌属(Clostridium)、动胶菌属(Zoogloea)、管道杆菌属(Cloacibacterium)、脱硫弧菌属(Desulfovibrio)等具有反硝化功能的菌群为b的优势菌属。     

接种两株有益菌的生物絮团净水性能及饲料化潜力

为强化新的絮团形成菌伪杜擀氏菌YN12(Pseudoduganella eburnea YN12, YN)利用养殖尾水生成生物絮团效果,并将絮团制成饲料,探讨在胡子鲶(Claris fuscus)饲养上的可行性。在生物絮团系统中,接种活性污泥(Activated sludge, AS)、伪杜擀氏菌YN12和枯草芽孢杆菌(Bacillus subtilis, BS)并配制成7组(AS、YN、BS、AS+YN、AS+BS、YN+BS和AS+YN+BS),分析絮团形态结构差异,评估出对模拟养殖尾水氮去除效果及微生物群落结构优势的最佳组合,最终收集最佳组合絮团烘干研磨成粉末添加到商业饲料制粒,分成对照组和絮团组两组,探讨对胡子鲶的生长性能、饲料利用率、肌肉营养成分及肝脏的消化酶和免疫酶活性的影响。结果表明:YN+BS组水质净化效果稳定,总氮去除效果(89.6%)良好,氨氮、亚硝酸盐和硝酸盐积累低,具有良好的絮团生成量(9.0 ml/L)及占据优势(47.5%)的反硝化菌群(包括Hydrogenophaga、Flavobacterium、Pseudoxanthomonas、Burkholderi...     

蔗糖输入对凡纳滨对虾养殖系统真核微生物群落的影响

利用18S rRNA基因高通量测序, 研究蔗糖输入条件下凡纳滨对虾(Litopenaeus vannamei)养殖系统中水体与絮团的真核微生物群落组成及其相互作用关系。结果表明, 输入蔗糖能显著改变对虾养殖系统中真核微生物群落组成, 促进水体中纤毛虫和轮虫的生长, 对水体绿藻的过度生长有一定抑制作用。Mantel分析发现, 在第28天时, 蔗糖添加以及系统中的溶解氧、亚硝酸盐和磷酸盐对真核微生物群落结构变异有显著影响。利用共线性网络分析表明碳氮比的增加可以增强水体和絮团中不等鞭毛类、纤毛虫和轮虫的相互作用, 对养殖体系中藻类的过度繁殖具有一定的抑制作用。研究揭示了在蔗糖输入条件下凡纳滨对虾养殖系统中真核微生物群落的演替规律及絮团和水体微生物相互作用特征, 为生物絮团技术持续稳定的应用于对虾养殖系统提供理论支持。     

基于盲肠菌群和回肠粘膜基因表达探讨解淀粉芽孢杆菌提高蛋鸡生产性能的潜在机制

【背景】我们的前期工作表明,在蛋鸡饲料中添加解淀粉芽孢杆菌BLCC1-0238可显著提高产蛋性能和蛋品质,但其益生机制还不清楚。【目的】从基因组学和转录组学的视角去揭示益生菌提高蛋鸡生产性能的潜在机制。【方法】利用高通量测序技术比较分析基础日粮组(C组)和解淀粉芽孢杆菌BLCC1-0238添加组(0.06%,2.0×1010 CFU/g,T组)盲肠微生物菌群和回肠粘膜基因表达的差异。【结果】基因组学分析发现,与对照组相比,益生菌添加组的盲肠菌群Chao1和Shannon指数升高;在门水平上,厚壁菌门/拟杆菌门(Firmicutes/Bacteroidetes)的比例显著增加,梭杆菌门(Fusobacteria)和变形菌门(Proteobacteria)的相对丰度降低;在属水平上,厚壁菌门中的考拉杆菌属(Phascolarctobacterium)、乳酸杆菌属(Lactobacillus)、瘤胃球菌科UCG-014属(Ruminococcaceae UCG-014)、厌氧棍状菌属(Anaerotruncus)、瘤胃梭菌属(Ruminiclostridium9)、克里斯滕森氏菌(Christensenellaceae_R-7_group)相对丰度明显增加。转录组学结果表明,回肠粘膜组织中共有356个差异表达基因,其中,上调基因199个,下调基因157个。生物信息学分析提示,上调基因参与了淀粉和蔗糖代谢、半乳糖代谢以及甘氨酸、丝氨酸和苏氨酸代谢等促进物质代谢和营养吸收的过程。【结论】日粮中添加解淀粉芽孢杆菌BLCC1-0238,可通过改善盲肠菌群多样性和肠道营养吸收能力提高蛋鸡的饲料利用率和产蛋性能,具有潜在的实用价值。     

养殖水质原位自净微生物诱导及复合应用

本文采用原位诱导有益微生物（污水菌、芽胞杆菌、硝化菌和副球菌等）,复合添加外源的有益微生物（金藻、栅藻、硅藻、芽胞杆菌、光合菌、酵母菌和EM（effective microorganisms）菌等）,在养殖水体中建立稳定的有益微生物复合菌群,使水体中亚硝酸氮和氨氮清除率分别可达100%和99%。并进一步利用这些菌群在小红鲫鱼（red crucian carp）和南美白对虾（Penaeus vannamei）养殖中进行原位水质净化应用。在小红鲫鱼养殖水质原位自净中,无需清污和换水,就可长期维持水环境完全稳定,连续养殖50 d以上,水中未检测出亚硝酸氮,氨氮低于0.2 mg/L,稳定在优质安全的养殖环境;在南美白对虾养殖中,投菌后初期水体亚硝酸氮较快下降,第10 d后氨氮浓度也降至养殖安全范围内,形成稳定安全的养殖水质环境。实验效果显著,表明微生物菌群原位水质自净技术具有推广应用前景。     

七氟菊酯和溴氰菊酯对棉铃虫肠道菌群的影响

【目的】本研究旨在探究拟除虫菊酯类杀虫剂对棉铃虫Helicoverpa armigera幼虫肠道菌群结构及代谢的影响,丰富对杀虫剂作用机理的认识。【方法】分别对棉铃虫2和3龄幼虫饲喂普通人工饲料(对照组, SS)、含2%七氟菊酯(Ⅰ型拟除虫菊酯)粉剂饲料(七氟菊酯处理组,Te)和含2.5%溴氰菊酯(Ⅱ型拟除虫菊酯)乳油饲料(溴氰菊酯处理组, DM),然后提取3龄幼虫肠道菌群基因组DNA;利用Illumina MiSeq二代高通量测序技术对肠道细菌的16S rDNA的V3-V4变异区进行测序,分析其肠道细菌的多样性和丰富度;利用qPCR验证16S rDNA测序分析结果。取2和3龄幼虫肠道,匀浆后进行Biolog-Eco实验,分析肠道细菌对Eco板上31种碳源的代谢情况。【结果】16S rDNA测序结果表明,棉铃虫3龄幼虫肠道细菌主要是厚壁菌门(Firmicutes)、变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)和蓝藻菌门(Cyanobacteria)。与对照组相比,溴氰菊酯处理组和七氟菊酯处理组的棉铃虫幼虫肠道细菌的α多样性指数没有显著性改变,但是菌群结构发生了变化：在门水平,拟杆菌门的相对丰度减少,厚壁菌门和蓝藻菌门的相对丰度增加,qPCR验证结果亦支持16S rDNA测序分析的这个结果;在属水平,拟杆菌属Bacteroides、普氏菌属Prevotella和假单胞菌属Pseudomonas等的相对丰度降低,狭义梭菌属Clostridium sensu stricto 1、埃希菌属-志贺氏菌属Escherichia-Shigella和盐单胞菌属Halomonas等的相对丰度增加,其中盐单胞菌属Halomonas的相对丰度显著增加。Biolog-Eco结果表明,与对照组相比,溴氰菊酯处理组中2龄幼虫对羧酸类碳源的代谢能力下降;溴氰菊酯处理组和七氟菊酯处理组中3龄幼虫对DL-α-磷酸甘油、肝糖和L-苯丙氨酸等碳源的利用能力下降。【结论】结果显示,拟除虫菊酯类杀虫剂对棉铃虫肠道菌群的结构和代谢能力有明显影响,拟除虫菊酯类杀虫剂使棉铃虫肠道有益菌的相对丰度下降,而使致病菌的相对丰度增加。短时间拟除虫菊酯处理未造成抗药性菌群的丰度增加。qPCR检测结果与16S rDNA测序分析结果相似。Ⅰ型和Ⅱ型拟除虫菊酯类杀虫剂对棉铃虫肠道菌群结构和代谢功能的影响不同。     

基于高通量测序的缢蛏及其养殖池塘菌群结构的季节变化

为探究缢蛏养殖过程中养殖环境和缢蛏体内的细菌群落结构季节变化特征及其与环境因子的相关性,采用高通量测序技术对缢蛏养殖池塘的水体、沉积物及缢蛏内脏团细菌菌群进行了研究.结果表明: 水体中冬季的细菌菌群结构与其他3季差异显著,沉积物和缢蛏内脏团4季的细菌菌群结构均无显著差异.水体各季节细菌群落的Shannon多样性指数无明显差异,沉积物及缢蛏内脏团细菌群落Shannon多样性指数夏季最低,冬季最高.门水平上,蓝细菌门、变形菌门和拟杆菌门为水体的优势菌群;变形菌门、绿弯菌门和拟杆菌门为沉积物的优势菌群;柔膜菌门和变形菌门为缢蛏内脏团的优势菌群.属水平上,水体冬季的优势菌属为NS3a_marine_group,而其余3季为聚球藻属;沉积物优势菌属为厌氧绳菌科下的一类菌属norank_f_Anaerolineacea和硝化螺旋菌属;内脏团优势菌属为支原体属和弓形菌属.环境因子与菌群的相关性分析表明: 水体的优势菌属聚球藻属与水温、化学需氧量(COD)、PO₄^--P、NH₄⁺-N、pH和透明度呈正相关;沉积物的优势菌属厌氧绳菌科菌属norank_f_Anaerolineacea与COD、水温、总磷呈正相关;缢蛏内脏团的优势菌属支原体属与水温、pH、NH₄⁺-N、PO₄^--P和透明度呈正相关.这表明不同季节的缢蛏养殖池塘及缢蛏体内细菌群落结构及多样性差异较大,水体细菌菌群受养殖环境的影响明显,尤其是水温及氮磷含量.     

外加碳源对蚯蚓-污泥系统细菌群落结构的影响

蚯蚓是土壤生态系统中有机物质的重要分解者,并可通过物理运动和排泄蚓粪改变土壤的微生物群落组成。为研究蚓粪与污泥中微生物的相互关系,利用赤子爱胜蚓(Eisenia foetida)和人工湿地基质构建蚯蚓-污泥系统,采用高通量测序技术比较分析添加葡萄糖、乳糖、淀粉、纤维素4种碳源后污泥和蚓粪的细菌多样性及群落结构变化。结果表明,污泥的Chao和ACE指数明显高于蚓粪,添加不同碳源实验组间污泥的Chao和ACE指数差别不大,但均高于污泥对照组;添加葡萄糖、淀粉和纤维素实验组间蚓粪的Chao和ACE指数相近且高于蚯蚓对照组,乳糖实验组蚓粪的Chao和ACE指数则明显低于蚓粪对照组。添加葡萄糖、乳糖、淀粉和纤维素能够增加污泥中细菌多样性;添加葡萄糖、淀粉和纤维素能够增加蚓粪中细菌多样性,但添加乳糖会导致蚓粪中细菌多样性降低。蚓粪与污泥细菌群落结构组成存在明显差异,污泥中变形菌门、拟杆菌门、厚壁菌门、放线菌门、绿弯菌门、酸杆菌门、芽单胞菌门为优势菌门;除乳糖组外,蚓粪中变形菌门、拟杆菌门、厚壁菌门、放线菌门、疣微菌门为优势菌门(其中变形菌门和拟杆菌门占绝对优势),蚓粪乳糖组检测到柔膜菌门(2.6%),其他实验组均未检出。添加碳源后,污泥和蚓粪中能分解相应碳源的菌群增加,如葡萄糖实验组Lactococcus piscium,乳糖组Flavobacterium reichenbachii,淀粉组Alkanindiges illinoisensis和Zobellella taiwanensis,纤维素组Cellvibrio gandavensis等。     

凡纳滨对虾生物絮团养殖系统中不同生境细菌群落特征

【目的】凡纳滨对虾生物絮团养殖系统(biofloc-based culture system, BFS)是一种基于培育和调控微生物群落的新型生态养殖模式。然而,目前对于BFS在不同生境中的微生物群落特征及其构建过程还不清楚。【方法】采用16S rRNA基因测序技术探究BFS在3种不同生境(水体、絮团和对虾肠道)的细菌群落组成;通过溯源分析和中性模型等方法,探究不同生境细菌群落的特征及其构建过程。【结果】3种生境的微生物群落多样性和组成具有显著性差异,絮团和对虾肠道的群落结构和组成最为相似,溯源结果显示对虾肠道有98.76%的细菌类群来自絮团,仅有0.83%的细菌类群来自水体;3种生境共有的细菌主要为鲁杰氏菌(Ruegeria),在水体、絮团和对虾肠道中的丰度分别为1.72%、7.34%和6.00%,水体中特有的扩增子变异序列(amplicon sequence variants, ASV)数量为89个,主要属于海茎状菌(Maricaulis)和欧文威克斯菌(Owenweeksia),絮团中有56个,主要为莱茵海默氏菌(Rheinheimera),而对虾肠道中仅有10个,主要属于玫瑰杆菌(Roseobacter);中性模型结果表明,水体、絮团和对虾肠道细菌群落构建均符合中性模型,表明3种生境中细菌群落构建均受中性过程主导。【结论】在BFS系统中,不同生境的微生物群落具有显著差异,对虾肠道细菌主要来自生物絮团,而3种生境的细菌群落构建过程由中性过程主导。这些结果为调控生物絮团养殖系统中微生物群落提供了理论依据。     

一株光合细菌的分离鉴定及该菌对氨氮和亚硝态氮的去除作用

【背景】中国是水产养殖大国,氨氮、亚硝态氮是水体中主要的氮源污染物。水体氨氮超标不仅会损伤水生动物的神经系统和肝肾系统,还会导致体表及内脏充血。亚硝态氮过高会阻碍血液运载氧气能力,导致鱼虾缺氧、免疫力下降,从而引发肠炎、烂鳃,甚至窒息死亡。部分光合细菌有去除水体氨氮、亚硝态氮的能力,且对环境友好无二次污染。【目的】从广东养殖水体分离、纯化、筛选出生物活性好的光合细菌(编号SP3)进行种属鉴定,优化培养条件,检测其去除水体氨氮和亚硝态氮的能力,为养殖水体去除氨氮和亚硝态氮提供目标菌株。【方法】用双层平板法从混合菌液中分离得到光合细菌,通过革兰氏染色、碳源利用试验、对无机电子供体的利用试验以及16SrRNA基因序列分析对目标菌株进行种属鉴定;测定菌株SP3在不同pH、不同浓度NaCl条件下的OD600,优化培养条件;通过测定7d内SP3菌株在不同浓度氨氮(氯化铵配制)和亚硝态氮(亚硝酸钠配制)中OD600的变化趋势,确定该菌株对不同氮源的利用情况;用纳氏比色法、分光光度法测定SP3菌株降解水体氨氮和亚硝态氮的能力。Genome walking扩增获得亚硝酸盐还原酶基因(nirS),通过荧光定量PCR研究nirS在氨氮、亚硝态氮去除过程中的表达动态。【结果】筛选出的菌株SP3为革兰氏阴性菌,短杆状;能以醋酸盐、丙酮酸盐、丙酸盐、丁酸盐、乳酸盐、富马酸盐、琥珀酸盐、苹果酸、果糖、葡萄糖作为碳源,不能以乙醇和丙酮作为碳源;能利用硫化钠、硫代硫酸钠、亚硫酸钠作为无机电子供体; 16SrRNA基因序列分析表明其与沙氏外硫红螺菌(Ectothiorhodospira shaposhnikovii)序列相似度为99%;菌株SP3适宜pH为6.0-8.5,适宜盐度为0-3%;菌株SP3以铵盐作为氮源时生长状态明显优于亚硝酸盐;以初始菌液浓度8.6×109CFU/mL、起始氨氮量84.15±0.58 mg/L的条件培养7 d,水体氨氮累计去除量为79.45±0.29 mg/L,氨氮累计去除率达到94.42%;在同样菌浓度和2mg/L亚硝酸钠的条件下培养5d,水体亚硝态氮含量低于检测限0.003mg/L。在菌株SP3去除氨氮、亚硝态氮过程中nirS相对表达量上调。【结论】菌株SP3为沙氏外硫红螺菌(E.shaposhnikovii),能有效去除水中氨氮和亚硝态氮,具有净化水质作用,在水产养殖和污水处理中有广阔应用前景。     

不同碳氮比对生物絮团形成及对日本沼虾生长、抗氧化酶和消化酶的影响

为了研究不同碳氮比对生物絮团形成及对日本沼虾(Macrobrachium nipponense)生长、抗氧化酶和消化酶的影响,设置5个不同实验组[对照组(不做任何添加),碳氮比10组(C/N10)、碳氮比15组(C/N15)、碳氮比20组(C/N20)和碳氮比25组(C/N25)],每组设三重复;将初始体重(0.25±0.03) g的日本沼虾置于不同碳氮比的玻璃缸(30 cm×40 cm×100 cm)中,进行50d的饲养实验。研究结果表明,随着碳氮比升高,生物絮团含量有上升趋势,生物絮团含量C/N20>C/N25>C/N15>C/N10, C/N20生物絮团最多,生物絮团体积和总固体悬浮物分别为(328.67±7.09) mL/L和(40.33±1.53) mg/L而C/N10几乎没有生物絮团产生, C/N15和C/N25有少量生物絮团,但含量显著低于C/N20。对照组的氨氮和亚硝酸氮浓度持续升高, C/N10和C/N25的氨氮和亚硝酸氮浓度有较大波动,先升高后降低,随后又有升高的趋势,氨氮和亚硝酸氮浓度分别高于2和1.5 mg/L, C/N15和C/N20的氨氮和...     

海水和淡化水养殖凡纳滨对虾饲料蛋白需求量的比较研究

本研究从生长、饲料利用和体成分等指标综合评价饲料蛋白质含量对凡纳滨对虾的影响。研究发现,在海水养殖环境下,饲料蛋白水平对凡纳滨对虾的存活率、增重率、特定生长率、饵料系数、蛋白效率和虾体粗蛋白含量均有显著性影响(p<0.05)。摄食饲料蛋白含量38%的海养凡纳滨对虾有较高的存活率、增重率以及最高的特定生长率。饵料系数随蛋白含量的增加而减小,虾体粗蛋白含量随饲料蛋白水平升高而增加,而蛋白含量38%和41%实验组饵料系数和虾体粗蛋白含量均没有显著性差异(p>0.05)。淡化水养殖条件下,凡纳滨对虾的存活率随饲料蛋白水平的增加而提高,而增重率和特定生长率均在饲料蛋白含量35%实验组最高,饵料系数随蛋白含量的增加先减小后增大,蛋白含量35%实验组饵料系数显著小于其他实验组。海水养殖条件下,凡纳滨对虾的存活率、末体重、增重率、特定生长率和蛋白质效率均高于淡水养殖的对虾。对虾虾体粗蛋白、粗脂肪和粗灰分含量均随盐度升高而升高,而虾体水分随着盐度的升高而降低。     

一种复合微生态制剂对养殖水体中生物因子的影响

[目的]研究海水养殖池塘中投放以芽孢杆菌和假单胞菌为主的一种自主研发的微生态制剂对池塘养殖生物、微生物菌相、浮游生物的影响,以期为建立微生物生态调控技术体系提供参考.[方法]实验期间,每隔15d于试验池塘中泼洒微生态制剂一次,每月末取样测量水体和底泥中异养菌和弧菌密度、浮游生物种类和密度,实验结束时测量养殖生物的生长率和成活率.[结果]该微生态制剂能够显著提高南美白对虾和三疣梭子蟹的存活率和生长速度,试验组南美白对虾的存活率和体重增长率较对照组分别提高了11.3％和1 400％,试验组三疣梭子蟹的存活率和体重增长率较对照组分别提高了1.2％和37.5％;微生态制剂能够显著提高海水池塘的异养菌总数,从而抑制致病性弧菌的数量,试验组池水和底泥的异养菌平均密度在整个养殖期较对照组提高了58.5％和51.3％,而试验组水体中致病菌——弧菌的数量在整个养殖期较对照组降低了39.7％;微生态制剂使绿藻门和硅藻门的藻类密度明显增加,对蓝藻和甲藻的生长有抑制作用.[结论]该微生态制剂能够有效地改善养殖池塘中生物群落结构,提高某些养殖品种的生长速度.     

Administration of Bacillus subtilis strains in the rearing water enhances the water quality,growth performance,immune response,and resistance against Vibrio harveyi infection in juvenile white shrimp,Litopenaeus vannamei

In this study, vegetative cell suspensions of two Bacillus subtilis strains, L10 and G1 in equal proportions, was administered at two different doses 10⁵ (BM5) and 10⁸ (BM8) CFU ml⁻¹ in the rearing water of shrimp (Litopenaeus vannamei) for eight weeks. Both probiotic groups showed a significant reduction of ammonia, nitrite and nitrate ions under in vitro and in vivo conditions. In comparison to untreated control group, final weight, weight gain, specific growth rate (SGR), food conversion ratio (FCR) and digestive enzymatic activity were significantly greater in the BM5 and BM8 groups. Significant differences for survival were recorded in the BM8 group as compared to the control. Eight weeks after the start of experiment, shrimp were challenged with Vibrio harveyi. Statistical analysis revealed significant differences in shrimp survival between probiotic and control groups. Cumulative mortality of the control group was 80%, whereas cumulative mortality of the shrimp that had been given probiotics was 36.7% with MB8 and 50% with MB5. Subsequently, real-time RT-PCR was employed to determine the mRNA levels of prophenoloxidase (proPO), peroxinectin (PE), lipopolysaccharide- and β-1,3-glucan- binding protein (LGBP) and serine protein (SP). The expression of all immune-related genes studied was only significantly up-regulated in the BM5 group compared to the BM8 and control groups. These results suggest that administration of B. subtilis strains in the rearing water confers beneficial effects for shrimp aquaculture, considering water quality, growth performance, digestive enzymatic activity, immune response and disease resistance.     

Performance of a constructed wetland in treating brackish wastewater from commercial recirculating and super-intensive shrimp growout systems

A recirculating aquaculture system was developed for treating Pacific white shrimp (Litopenaeus vannamei) production wastewater using an integrated vertical-flow (IVF) and five connected integrated horizontal flow (IHF) constructed wetlands as water treatment filters for mesohaline conditions (8.25‰-8.26‰ salinity). The constructed wetlands demonstrated the ability to reduce total nitrogen, total ammonia nitrogen, nitrite nitrogen, nitrate nitrogen, total phosphorous, chemical oxygen demand, and total suspended solids to levels significantly lower than those in effluents from culture tanks. Various water quality parameters in the culture tanks were deemed suitable for shrimp culture. The actual ratio of wetland area (A(w)) to culture tank area (A(t)) was 1.1439, and the estimated optimal ratio A(w)/A(t) was approximately 1. The IVF-IHF wetlands showed flexibility and reliability in consistently removing the main pollutants from commercial recirculating and super-intensive shrimp growout systems throughout the culture period.     

Responses to dietary starch or sucrose on protein and glucose kinetics in goats fed a high-concentrate diet

Responses of whole body protein synthesis (WBPS) and glucose irreversible loss rate (ILR) were compared between dietary starch and sucrose in four male goats. Diets were fed at 1.2 times maintenance requirements of ME and CP with 30% of the ME as starch, starch plus sucrose or sucrose, twice daily. The diets consisted of 33, 32, 11 and 24% of alfalfa hay, corn, soybean meal and the carbohydrates, respectively. The WBPS and glucose ILR during 5-7 h after feeding were determined by an isotope dilution method of [2H5]phenylalanine, [2H2]tyrosine, [2H4]tyrosine and [13C6]glucose. Sucrose elevated ammonia nitrogen and lowered acetate concentrations in the rumen, but did not differ from starch in nitrogen retention. Glucose ILR and WBPS were similar between the carbohydrates. It was concluded that dietary sucrose would have effects similar to starch on WBPS and glucose kinetics in the absorptive state in goats fed a high-concentrate diet.     

Development of biosensors for the simultaneous determination of sucrose and glucose, lactose and glucose, and starch and glucose

Sensors for the simultaneous determinations of sucrose and glucose, lactose and glucose, and starch and glucose were prepared by a combination of the enzyme system shown below and an oxygen electrode: The mechanism for separating the substrates with the proposed sensors is based on the time lag arising from reaction and diffusion. Invertase, beta-galactosidase, amyloglucosidase, mutarotase, and glucose oxidase were covalently immobilized on triacetyl cellulose membranes containing 1,8-diamino-4-aminomethyloctane. A glucose oxidase membrane, mutarotase membrane, three sheets of triacetyl cellulose membranes, and invertase, or beta-galactosidase or amyloglucosidase membrane were placed in that order on the tip of the oxygen electrode. Calibration curves for sucrose, lactose, and starch were linear up to 40 mM, 60-180 mM, and 10%, respectively. The simultaneous determination of sucrose and glucose, lactose and glucose, and starch and glucose was possible when the amount of glucose coexised was in the range of 2-16% sucrose, 2.8-8.3% lactose, or 0.1-1% starch. The relative errors were +/-4% for sucrose and +/-3% for lactose in 100 assays. The starch sensor was reused only five times. Each enzyme membrane was fairly stable for more than 10 days.     

Bacterial Community Dynamics During Nursery Rearing of Pacific White Shrimp ( Litopenaeus vannamei ) Revealed via High-Throughput Sequencing

A 20-day trial was conducted to reveal bacterial community dynamics in a commercial nursery of larval Litopenaeus vannamei larvae. The bacterial communities in the ambient water were profiled by high-throughput sequencing of the V4–V5 hypervariable region of the 16S rRNA gene. The results indicated that the dominant bacterial phyla between the metamorphosis stage and postlarval stage were Bacteroidetes, Proteobacteria, Cyanobacteria, and Firmicutes, representing more than 80.09% of the bacterial operational taxonomic units. The relative abundance among bacterial phyla notably differed between the two stages. The relative abundance of Cyanobacteria was higher in the metamorphosis stage, while that of Bacteroidetes was higher and more stable in the postlarval stage. At the class level, the relative abundance of Sphingobacteriia and Alphaproteobacteria increased markedly in the postlarval stage, while that of Flavobacteriia decreased. Redundancy analysis showed that bacterial composition in the metamorphosis stage was positively correlated with salinity, alkalinity, and pH, while in the postlarval stage, it was positively correlated with ammonium nitrogen and nitrite nitrogen. Thus, microbial community diversity in the nursery phase varies per rearing stage.