中草药复合益生菌制剂对凡纳滨对虾生长、抗病力及水质的影响

将枯草芽孢杆菌、纳豆芽孢杆菌、地衣芽孢杆菌等比例混合,制成芽孢杆菌制剂;将芽孢杆菌制剂与粪肠球菌、嗜酸乳杆菌等比例混合制成复合益生菌制剂;将分别用3种芽孢杆菌发酵的中草药等比例混合制成中草药芽孢杆菌制剂;将分别用5种益生菌发酵的中草药等比例混合,制成中草药复合益生菌制剂。在饲料中分别添加4种益生菌制剂(活菌为2×10~7 cfu/g),研究4种制剂对凡纳滨对虾(Litopenaeus vannamei)生长、抗病力及水质的影响。结果表明:1)4种益生菌制剂均可提高凡纳滨对虾的成活率、增重率和饲料利用率(P0.05),中草药益生菌制剂组的促生长效果优于益生菌制剂组,以中草药复合益生菌制剂组促生长效果为最佳(P0.05);2)4种益生菌制剂均可维持对虾养殖水体p H值、氨态氮和亚硝酸盐含量的稳定(P0.05),中草药益生菌制剂对水质的改良效果优于益生菌制剂(P0.05);3)用1×10~8 cfu/m L的哈维氏弧菌(Vibro harveyi)菌液浴浸泡凡纳滨对虾10 d,凡纳滨对虾的累计死亡率由大到小依次为中草药复合益生菌制剂组(31.11%)、复合益生菌制剂组(35.56%)、中草药芽孢杆菌制剂组(37.78%)、芽孢杆菌制剂组(44.44%)、对照组(93.33%)。在饲料中添加一定比例的中草药复合益生菌制剂可提高凡纳滨对虾的生长指标、改善养殖水体环境、提高对虾的抗病力。     

几株有益芽孢杆菌对温度、制粒工艺及pH值的耐受性

报道了实验室分离、筛选的 5株有益芽孢杆菌对温度、对虾饲料制粒工艺流程和pH值的耐受性。结果表明 ,5株芽孢杆菌经 80℃水浴 4 0min后全部存活 ,90℃水浴 2 0min有 35 %～ 70 %存活 ,10 0℃水浴 10min仍有 30 %～ 5 0 %存活 ,显示所筛选的 5株芽孢杆菌对高温有较强的耐受力 ;在对虾饲料中添加 5株芽孢杆菌 ,经整个生产工艺流程后芽孢杆菌存活 95 %,烘干后芽孢杆菌存活 93%,说明杆菌能够承受饲料制粒生产中压力、温度和水分的变化 ;当pH值为 3.8、4 .6、5 .2时 ,分别有 1、3、5株菌能繁殖 ,pH值 6 .0～ 8.5时 ,5株菌生长旺盛 ,将 5株菌经pH值 2 .2～ 4 .6处理1h ,再接种在pH值 7.2的培养基可以良好生长 ,说明杆菌在酸性环境中生长性能较差 ,在弱酸性和碱性环境中生长旺盛 ;能耐受酸性环境而存活 ,再给予适宜环境即能良好生长。 5株芽孢杆菌具有作为水产硬颗粒饲料专用益生素菌株的优势。     

几株有益芽孢杆菌对温度、制粒工艺及pH值的耐受性

报道了实验室分离、筛选的5株有益芽孢杆菌对温度、对虾饲料制粒工艺流程和pH值的耐受性。结果表明，5株芽孢杆菌经80℃水浴40min后全部存活，90℃水浴20min有35％-70％存活，100℃水浴10min仍有30％-50％存活，显示所筛选的5株芽孢杆菌对高温有较强的耐受力；在对虾饲料中添加5株芽孢杆菌，经整个生产工艺流程后芽孢杆菌存活95％，烘干后芽孢杆菌存活93％，说明杆菌能够承受饲料制粒生产中压力、温度和水分的变化；当pH值为3．8、4．6、5．2时，分别有l、3、5株菌能繁殖，pH值6．0-8．5时，5株菌生长旺盛，将5株菌经pH值2．2—4．6处理1h，再接种在pH值7．2的培养基可以良好生长，说明杆菌在酸性环境中生长性能较差，在弱酸性和碱性环境中生长旺盛；能耐受酸性环境而存活，再给予适宜环境即能良好生长。5株芽孢杆菌具有作为水产硬颗粒饲料专用益生素菌株的优势。     

有益微生物改善养殖生态研究──Ⅰ.复合微生物分解有机底泥及对鱼类的促生长效应

应用以芽孢杆菌为主导菌的微生物复合制剂进行分解养鱼池有机污泥的试验,菌剂合活菌数为109个／g．用量为1.5～4．5mg·L－1。经一个月的试验,池底原有厚3～5cm的有机污泥被分解,并对鱼类有明显的促生长作用。说明应用以芽孢杆菌为主导菌的微生物复合制剂净化养殖池塘底质是一种有效的措施。     

基于PCR和TOF-MS海洋源产脂肽芽孢杆菌筛选

【目的】准确高效分离红树林等海洋环境中产脂肽芽孢杆菌。【方法】将前期从红树林环境筛出具有抗菌活性的12株芽孢菌株作为研究菌株,采用溶血活性及排油作用对其中的可能产脂肽菌株进行初步筛选,并基于脂肽合成酶基因PCR扩增法对脂肽合成酶基因携带菌复筛,采用飞行时间质谱(TOF-MS)对所产脂肽进行鉴定。【结果】初步发现6株芽孢杆菌具有溶血活性,其中HY-5、HY-4和HN-9这3株具有排油能力,油层透明圈的直径分别为45、32和3 mm。PCR扩增检测到HY-5含有iturin、surfactin和bacillomycin D合成酶基因,HY-4含有iturin、surfactin合成酶基因,HN-9含有iturin合成酶基因。TOF-MS检测到HY-5和HY-4的发酵液含surfactin、iturin和bacillomycin 3种脂肽,HN-9的发酵液只含iturin。【结论】基于PCR与TOF-MS为核心的级联技术,建立一种海洋源脂肽产生菌快速筛选以及脂肽谱快速鉴定的方法,解决海洋源芽孢杆菌在常规培养基难合成脂肽造成的漏筛情况。     

大口黑鲈肠道益生菌的分离与鉴定

【目的】分离鉴定大口黑鲈（Micropterus salmoides）肠道益生菌,评估分离菌用作饲料添加剂的益生潜力。【方法】采集大口黑鲈肠道样本,采用稀释涂布法分离细菌,经16S rRNA序列比对和生化试验鉴定种属,并进行抗逆性能、黏附能力、溶血活性和药敏特性等鉴定。【结果】从大口黑鲈肠道分离菌株84株,经测序、blast比对,筛选出3株同源性最高的菌株MS-1、MS-2、MS-3,分别鉴定为地衣芽孢杆菌（Bacillus licheniformis）、解淀粉芽孢杆菌（Bacillus amyloliquefaciens）和贝莱斯芽孢杆菌（Bacillus velezensis）。3株分离菌均为γ-溶血,不具有生物膜,疏水性50%～93.3%,自凝集性43.6%～69.2%;可在pH 2.0～12.0环境和5.0 g/L胆盐中存活;经高温处理后仍可生长;3株菌对大多数抗生素敏感。【结论】芽孢杆菌MS-1、MS-2和MS-3抗逆性高,黏附性能及饲料学安全性高,可作为大口黑鲈养殖中潜在益生菌。     

养殖场底泥中芽孢杆菌属细菌的生态学研究

对近海养殖场底泥细菌的研究 ,发现养殖池底泥中芽孢菌为优势菌群。应用形态学、生理生化和 16SrDNA序列测定 ,对芽孢杆菌属细菌做进一步分类的结果表明 ,该属细菌在底泥中的主要种群为蜡状芽孢杆菌、巨大芽孢杆菌、短小芽孢杆菌、坚强芽孢杆菌、Bacillusbaekryungensis等。同时 ,还分析了这些芽孢杆菌属细菌在底泥元素的地球化学循环中的作用 ,认为这类细菌应属于陆生微生物 ,在养殖环境中代谢有机化合物产生氨、亚硝酸盐和H2 S ,对养殖环境产生不利的作用和影响     

养殖场底泥中芽孢杆菌属细菌的生态学研究

对近海养殖场底泥细菌的研究，发现养殖池底泥中芽孢菌为优势菌群。应用形态学、生理生化和16S rDNA序列测定，对芽孢杆菌属细菌做进一步分类的结果表明，该属细菌在底泥中的主要种群为蜡状芽孢杆菌、巨大芽孢杆菌、短小芽孢杆菌、坚强芽孢杆菌、Bacillus baekryungensis等。同时，还分析了这些芽孢杆菌属细菌在底泥元素的地球化学循环中的作用，认为这类细菌应属于陆生微生物，在养殖环境中代谢有机化合物产生氨、亚硝酸盐和H2S，对养殖环境产生不利的作用和影响。     

过热蒸汽对熟制小龙虾优势腐败菌的杀菌动力学及其机理

【目的】研究过热蒸汽杀菌对熟制小龙虾优势腐败菌的杀菌效果。【方法】从腐败熟制小龙虾中分离腐败菌,根据16S rDNA基因序列信息鉴定其种属,研究过热蒸汽温度、流量和杀菌时间等的处理参数对小龙虾优势腐败菌的杀菌效果,并利用Weibull模型拟合其杀菌动力学过程。通过研究过热蒸汽处理对优势腐败菌菌悬液的电导率和紫外吸收的影响,以及通过扫描电子显微镜对菌体的微观结构进行观察,初步探讨其杀菌机理。【结果】熟制小龙虾优势腐败菌为短小芽孢杆菌(Bacillus pumilus)、解淀粉芽孢杆菌(Bacillus amyloliquefaciens)、蜡样芽孢杆菌(Bacillus cereus)和赖氨酸芽孢杆菌(Lysinibacillus macroides)。在过热蒸汽杀菌初期阶段,优势腐败菌的残活率迅速下降,随着杀菌时间延长,残活率下降速率变得缓慢,最后趋于稳定,说明过热蒸汽能完全杀灭小龙虾优势腐败菌。Weibull模型能较好拟合过热蒸汽对4种优势腐败菌的杀菌动力学,其R2≥0.935。Weibull模型得到的预测值与实测值接近程度较高。通过对模型的分析,提高过热蒸汽温度和增大流量,更容易到达灭活腐败菌的效果。过热蒸汽杀菌后,菌悬液的电导率值和吸光度迅速上升,之后趋于平缓;通过对微观结构的观察,发现杀菌后腐败菌的细胞膜完整性遭到严重破坏。【结论】过热蒸汽杀菌技术应用于小龙虾产品加工具有一定的可行性。     

马氏珠母贝育珠细胞小片和珠核处理技术研究Ⅲ:插核手术贝伤口区弧菌类型的初步研究

从马氏珠母贝(Pinctada martensii)插核部位蘸取组织液分离弧菌,研究插核手术前后室内休养3 d时伤口部位弧菌类型的变化,比较使用不同小片和珠核处理液对手术伤口部位弧菌类型的影响。形态和生理生化特征分型和鉴定结果显示,插核前和休养期贝样上分离的优势弧菌类型无明显差异,只是休养期弧菌的分离频率有所下降;从术后不同处理组的马氏珠母贝伤口上均分离得到7个类型弧菌,弧菌所属的种类也一致,而从未插核的对照组相应部位上只分离到4个类型弧菌,除未分离到副溶血弧菌(Vibrio parahaemolyticus)外,对照组与两个处理组贝样上分离到的弧菌种类基本上一致。     

有效微生物菌群对养虾水体细菌生态和水质的影响

研究有效微生物菌群(EM)对养虾水体中异养细菌、弧菌、光合细菌、放线菌的数量以及水体水质的影响。结果表明,在养殖池中添加EM,对水体中的异养细菌、弧菌、光合细菌及放线菌的数量均有显著影响:实验组异养细菌数量比传统对照池减少40.0%,弧菌数量减少7.8%,光合细菌数量增加98.2%,放线菌数量增加99.2%。EM能改良水质条件,与对照组相比,实验组溶解氧提高11.0%,COD降低8.0%,氨氮含量降低20.7%,亚硝酸氮含量降低10.0%。另外,实验组对虾成活率比对照组提高23.08%,饵料系数下降9.4%,利润提高了16.11%。     

Heterotrophic bacterial floras in industrial area and unpolluted marine environments around Qingdao

From Oct. 1999 to Oct. 2000, the heterotrophic bacterial floras in the industrial marine environment around the Qingdao Power Plant (QPP) and in the unpolluted marine environments were investigated. The results showed that the numbers of the heterotrophic bacteria around QPP were much higher than those in unpolluted environments, and the average numbers in QPP Seawater, QPP Sediment, Unpolluted Seawater and Unpolluted Sediment were 5.4×10⁴cfu(mL)⁻¹, 5.0×10⁵cfug⁻¹, 3.0×10²cfu(mL)⁻¹ and 1.3×10⁵cfug⁻¹ respectively. Totally, 118 strains were isolated from QPP and 99 of them were Gram-negative. One hundred and twenty one strains were isolated from the unpolluted environments and 104 of them were Gram-negative. All the Gram-negative bacteria belonged to 13 genera. The distribution of the bacteria was varied in different marine environments. The results showed that the unpolluted marine environments contained much more Vibrio than seawater and sediment around QPP.     

Effect of stocking density on water quality and (Growth,Body Composition and Plasma Cortisol Content) performance of pen-reared rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>)

The goal of the study was to examine the effect of stocking density on the water quality of culture area, as well as the growth, body composition and cortisol content of rainbow trout（Oncorhynchus mykiss）. Pen-reared trout were stocked in densities of 40, 60, 80 fish individuals m^-3(4.6, 6.6, 8.6 kg m^-3, SD1, SD2 and SD3 groups, respectively) for 300 days. Compared to the water from SD1 and SD2, that from SD3 exhibited significantly higher NH₄⁺-N content and COD（chemical-oxygen-demand）, and a significant reduction of dissolved oxygen in day 180(40.6 kg m^-3). Stocking density was significantly associated with body weight, standard length, VSI（viscerosomatic index）, CF（condition factor） and FC（food coefficient） in group SD3, particularly in day 240 and day 300(45 or 49.3 kg m^-3). Increased crude fat and decreased crude protein were displayed in high density group when the density reached to 36 kg m^-3. As a cumulative effect of density-related stress, VSI, CF, FC, moisture, and crude protein content varied over time in each density group（SD1, SD2, and SD3）. In summary, trout exhibited a better growth performance in low density(26.3 kg m^-3) than those reared in high densities(36 and 45 kg m^-3). The results indicate that rainbow trout（114.44 g ± 6.21 g, 19.69 cm ± 0.31 cm） initially stocked in 6.6 or 8.6 kg m^-3 should be lightened to less than 36 kg m^-3 after an intensive rearing for 240 days.     

Soil properties in two soil profiles from terraces of the Nam Co Lake in Tibet, China

This article investigates the soil properties in two soil profiles from the terraces of the Nam Co Lake in Tibet, China. Profile 1, with a slope aspect to east, is 37 m, and profile 2, to southwest, is 32 m above the lake level. Twelve and nine layers were identified in profile 1 and profile 2, respectively, based on the color, texture, structure, moisture content, presence of carbonate, and root density. Grain sizes and measurements of water content, total organic carbon, total inorganic carbon, total carbon, pH value and electrical conductivity were all analyzed for each sample. The results show that soil is poorly developed because of sparse vegetation coverage and weak chemical weathering in the cold and arid area. The results also indicate that the soil properties are different between the two profiles and suggest that soil is better developed in profile 1 than in profile 2. These differences are ascribed to the dissimilarity of landform, vegetation and timing. Profile 1 faces to east as compared with profile 2 facing to southwest and gets less insolation and therefore less evaporation. Thus, vegetation grows better in profile 1 than in profile 2. Radiocarbon dating and OSL dating show that profile 1 underwent a longer soil forming process than profile 2, which is helpful for the soil development in profile 1.     

Microplastic pollution and enrichment of distinct microbiota in sediment of mangrove in Zhujiang River estuary,China

The microbial communities colonized on microplastics (MPs) have attracted widespread attention. However, few studies focused on the MPs impacts on mangrove ecosystems, particularly on bacterial communities. We investigated the MPs pollution in mangrove of Zhujiang (Pearl) River estuary (ZRE). To study the potential risk posed by MPs to the mangrove ecosystems, the differences in bacterial communities, functions, and complexity between MPs and sediment samples were reported for the first time. Microplastics (2 991±1 586 items/kg dry weight (dw)) in sediment were mainly fibers and polyethylene, mostly transparent, and in size less than 0.5 mm. Bacterial communities and functions significantly differed from MPs in mangrove sediment. Compared with sediment, MPs significantly enriched members of Proteobacteria, Bacteroidetes, and Actinobacteria, as well as the bacteria associated with plastic-degrading and human diseases on their surface, suggesting that microbial communities on MPs may promote MPs degradation and the spread of diseases, posing potential risk to mangrove ecosystems and human health. Although bacteria on MPs exhibited a lower diversity, the co-occurrence network analysis indicated that network of bacteria colonized on MPs was bigger and more complex than those of mangrove sediment, illustrating that MPs can act as a distinct habitat in this special ecosystem. This study provides a new perspective for increasing our understanding of microplastic pollution in mangrove ecosystems.

     

二氧化氯控制欧洲鳗细菌性疾病

以二氧化氯作为一种新型的消毒剂和杀菌剂 ,以传统含氯制剂 (如漂白粉、二氯异氰尿酸钠、三氯异氰尿酸 )作为对照 ,比较其在治疗欧洲鳗细菌性疾病及改善养殖水质方面的显著特性 ,其作用活性与环境 (如 p H、温度、氨及有机物 )因子的关系。结果表明 ,二氧化氯 (Cl O2 )对爱德华氏菌、嗜水气单胞菌和柱状屈挠杆菌有很好的杀灭效果 ,能迅速治愈欧洲鳗的赤鳍病和肝肾病 ,且能有效改善养殖水体 ,水中生化需氧量比对照组下降 10 %～ 30 % ,化学耗氧量下降 5% ,溶氧量提高 8%～ 10 %。当二氧化氯体积分数为 8m L/m3时 ,其杀菌率均高达 90 %以上 ,为氯制剂的 1.8倍 ,且活性不受环境因子的影响     

草鱼呼肠孤病毒vp5基因诱饵重组载体的构建、转化与自激活作用检测

根据草鱼呼肠孤病毒(Grass Carp Reovirus,GCRV)的vp5基因设计特异性引物,并扩增其开放阅读框序列(open reading frame,ORF),然后连接至pGBKT7载体上构建酵母双杂交诱饵载体pGBKT7-vp5,转化至酵母菌Y2HGold,并进行PCR鉴定以及自激活性和毒性检测。PCR、酶切以及测序表明,pGBKT7-vp5重组诱饵载体构建成功。自激活性和毒性检测显示,阳性重组菌株pGBKT7-vp5/Y2HGold和阴性对照菌株pGBKT7/Y2HGold在SD/-Trp平板上出现大小相一致的乳白色菌落,且在SD/-Trp/X-α-Gel、SD/-Trp/AbA+/X-α-Gel平板上生长出蓝色菌落,而在SD/-Trp/-Ade/-His无菌落出现。表明重组菌株pGBKT7-vp5/Y2HGold表达的融合蛋白激活了报告基因AUR1-C和MEL1,没有激活报告基因ADE2和HIS3并且对宿主菌无毒性。该重组诱饵载体可用于酵母双杂交系统进一步筛选cDNA文库中与其相互作用的蛋白。     

工厂化养殖暗纹东方鱼屯致病菌的分离和鉴定

从不同养殖场患病暗纹东方鱼屯(Takifaguobscurus)的鳃、肠、肝、肌肉及溃疡部位分离到9株菌株,经人工感染致病性试验,其T-22、T-11、J-2、H-3、H-5表现出较强的致病性,出现症状与自然发病鱼相同。通过对菌体形态特征、培养特性和API20E细菌生化快速鉴定试条的测定,分别鉴定为温和气单胞菌(Aeromonassobria)、豚鼠气单胞菌(Aeromonascaviae)、嗜水气单胞菌(Aeromonashydrophila)、类志贺邻单胞菌(Plesiomonasshigelloides)、霍乱弧菌(Vibriocholerae)。证实这5株菌为致病菌。药敏试验结果表明9种药物对温和气单胞菌、豚鼠气单胞菌、霍乱弧菌均有较好的抑菌效果;嗜水气单胞菌表现出明显的耐药性。氟嗪酸、环丙沙星、复合磺胺对类志贺邻单胞菌有一定抑菌效果。     

湛江市腌制金线鱼的微生物及理化指标检测

按照相关国家标准,分析湛江市腌制金线鱼(Nemipterus virgatus)中的微生物和理化指标.结果表明:腌制金线鱼的微生物安全指标中有3组样品菌落总数略超标,其他指标符合西班牙等国家腌渍鱼制品微生物标准;耐盐菌数为菌落总数20倍以上,优势耐盐菌全为革兰阴性杆菌,6株优势嗜盐菌中有2株为革兰阳性球菌;理化指标中挥发性盐基氮含量均低于300 mg/kg,符合水产品质量要求;甲醛及亚硝酸盐含量均低于5 mg/kg,相对较低.可见,湛江市腌制金线鱼安全质量状况较好     

Effects of different enzymatic hydrolysis methods on the bioactivity of peptidoglycan in Litopenaeus vannamei

The effects of different hydrolysis methods on peptidoglycan(PG) were assessed in terms of their impact on the innate immunity and disease resistance of Pacific white shrimp,Litopenaeus vannamei.PG derived from Bifidobacterium thermophilum was prepared in the laboratory and processed with lysozyme and protease under varying conditions to produce several different PG preparations.A standard shrimp feed was mixed with 0.05% PG preparations to produce a number of experimental diets for shrimp.The composition,concentration,and molecular weight ranges of the soluble PG were analyzed.Serum phenoloxidase and acid phosphatase activity in the shrimp were determined on Days 6-31 of the experiment.The protective activity of the PG preparations was evaluated by exposing shrimp to white spot syndrome virus(WSSV).Data on the composition of the PG preparations indicated that preparations hydrolyzed with lysozyme for 72 h had more low-molecular-weight PG than those treated for 24 h,and hydrolysis by protease enhanced efficiency of hydrolysis compared to lysozyme.SDS-PAGE showed changes in the molecular weight of the soluble PG produced by the different hydrolysis methods.Measurements of serum phenoloxidase and acid phosphatase activity levels in the shrimp indicated that the PG preparations processed with enzymes were superior to the preparation which had not undergone hydrolysis in enhancing the activity of the two serum enzymes.In addition,the preparation containing more low-molecular-weight PG enhanced the resistance of the shrimp to WSSV,whereas no increased resistance was observed for preparations containing less low-molecular-weight PG.These findings suggest that the immunity-enhancing activity of PG is related to its molecular weight and that increasing the quantity of low-molecular-weight PG can fortify the effect of immunity enhancement.