一种生物净水剂对河蟹养殖水体水质净化效果研究

The biological cleaning agent Pondplus was dowsed each ten days at different dosage into breeding ponds for Chinese mitten crab with stocking rate at 12 000 ind./ha, to evaluate its purification effects on water environment in breeding ponds and its effects on Chinese mitten crab growth. The results showed that dousing treatment improved the water quality significantly as evaluated according to the parameters of DO, CODMn, NH4 ＋ - N and NO2 - -N. Particularly, when the air temperature was higher than 35℃, the diurnal observation on DO values at the bottom of ponds showed that dousing at 450 g/（ ha•m） gave a DO value higher than 4 mg/L all day long, which was 2 mg/L higher than that of control. The observed increase of Chinese mitten crab were 14. 9% , 27.8% , 27.8% by average weight, when dousing at 150, 300, 450 g/（ha • m） , respectively. Accordingly, the production increased by 8.5%, 11.9%, 18.6% through its promotion of Chinese mitten crab growth, and the economic efficiency were enhanced by 96.2% , 159.8% , 180.9%.     

盐度对黄鲷胚胎发育及早期仔鱼生长的影响

The effect of salinity on embryo and early larva development of Dentex tumiforns was discussed. Floating forms of egg in different salinities, optimal salinity for embryonic development and early larva growth was studied. The results showed: 1. In immobile condition, all eggs sank at salinity of 32.0, most of egg ssuspended in the middle of water at 34.0, and all eggs floated on the surface when salinity above 36.0. 2. Eggs did not hatch out at salinity of below 10.0 or above 60.0 and the death time of egg gradually moved up with increase or decline of salinity. Eggs hatched out at salinity range of 15.0 and 50.0, including abnormal larva. There was indistinct difference in speed of embryonic development (about 36-40h) within the salinity of 15.0 and 50.0. However the salinity had a great effect on larval survival after hatching and abnormal rate. The relation between hatching rate and salinity variation showed parabola but abnormal rate showed inverted parabola. The hatching rate was 81%-86% and abnormal rate of yolk sac larva was 27%-30% in suitable salinities of 27.0-39.0. The hatching rate was 89%-91% and abnormal rate was 13%-16% in optimal salinities of 33.0-36.0. The oil ball of abnormal larva was located in the central or front position. With increased extent of salinity, spondyle of larva bended and number of larva with arrhythmia increased. 3. The optimal salinities were 30.0-35.0 based on SAI. The test with SAI showed that SAI of early larvae was 41.25-47.53 at salinities of 30.0-35.0, the yolk and oil ball of 7-8 days larva was completely absorbed with a survival rate of 88%.     

ABSTRACTS AND KEY WORDS

Added bagasse and Bacillus sp. to Litopenaeus vannamei intensive culture tank, the treatment groups were treatment A（added Bacillus sp.）, treatment B（added Bacillus sp. with crushing bagasse） and treatment C（added Bacillus sp. with crushing-steaming bagasse） respectively. To estimate the effect of addition of bagasse with Bacillus sp. on the culture environment and shrimp growth, the level of total ammonia nitrogen（TAN）, nitrite nitrogen and nitrate nitrogen, total amount of bacteria in the water and shrimp growth were determined. The result suggested that the TAN concentration in treatment B and C were significantly lower than that in treatment A during the early stage of the culture（P〈0.05）. The addition of bagasse with Bacillus sp. was contribute to the formation of bio-floccules. The concentration of bio-floccules in treatment B and Cwere 6.3-20 ml/L and 8.3-30 ml/L respectively, which were significantly higher than that in treatment A（2.7-8.3 ml/L） during the culture（P〈0.05）.The average weight of shrimp in treatment B and C were 8.56±0.21g and 8.84±0.26g respectively, which were significantly bigger than that in treatment A（7.66±0.40g） when harvest（P〈0.05）.     

大菱鲆胚胎的玻璃化冷冻保存

Vitrification solutions were examined for their suitability of cryopreservation of turbot (Seophthulnms maximus) embryos. PMPI gave higher survival rate (75.68%) than other vitrification solutions and was considered to be suitable liar the cryopreservation of turbot embryos. The freezing point of the vitrification solutions consisted of PG and MeOH in the proportion of 3:2 was measured when the vitrification solutions were cryopreserved. It showed that the vitrification solutions with a cryoprotectant concentration of over 41% have no freezing point and the freezing points of vitrification solutions containing 33.33%-40% cryprotectants were between -32.4℃ and -65.4℃， The freezing point decreased with the increase of cryoprotectant concentration. The resistance of turbot embryos at different stages to the cryoprotectants was studied, It was found that the turbot embryos from 4-5 pairs somite to tail bud were more resistant to cryoprotectants and suitable to vitrify. A live embryo was obtained after cryopreservation in liquid nitrogen for 14h and was hatched out。     

饥饿胁迫对鲤肠道与肝脏蛋白酶和淀粉酶活性的影响

The aim of the present study was to determine the effect of starvation on digestive enzymes activity of the carp （Cyprinus carpio） at the water temperature 18 -20℃ in the cycle and flow water aquarium. The experiment was designed for four groups with three replication,which were 0 day（control） , 10 days, 20 days, 30days group. The results showed That there was a trend to drop first and keep stabilization for Cyprinus carpio body weight and Meat Content with the prolongatin of starvation, and that Effect of starvation stress on the the activity of protease in intestine and hepatopancreas of Cyprinus carpio was significant, with the prolongatin of starvation, there was a trend to increase then drop for the activity of protease in fore-gut, mid-gut hind-gut and hepatopancreas of Cyprinus carpio. And that the variatin for activityof amylase in fore-gut was significant butthat of the others was no significant.     

细角螺的繁殖生态条件及繁殖习性

Hemifusus ternatanus(Gmelin), a gastropoda living in the deep water areas of undertide,as well as a delicious seafood, was distributed mainly in Chinese southeast seas and Japanese seas near the coast. In this experiment Hemifusus ternatanus' breeding ecological conditions and its propagation habit were studied in order to provide an experimental foundation for the artificial breeding. The brood stocks of test animals collected from the waters in Taiwan Strait, and then were examinated their growing temperature， salinity, feeding and propagation habit in the laboratory. The re sults showed that the growing temperature for Hemifusus ternatanus ranges from 16 ℃ to 32 ℃, with an optimum temperature between 20 ℃ to 28 ℃ and between 23 ℃ and 28 ℃ for breeding. The optimum growth salinity was 18.3-32.3 though it much adapts to salinity from 13 to 35. They prefer to feed on bivalves particularly those with thin shells and byssusless. Hemifusus ternatanus is dioecious and fertilization finishes inside the body. It was called as egg vesicle procreation that the whole stages of embryo growth were developed within egg vesicle. Larva forms as soon as it leaves egg vesicle, which is called direct occurrence type. The larva became the juvenile after metamorphosis during 20-25 days development. It was first to obtained 233 juveniles with shell height 32-45 mm by an artificial breeding method.     

鳗源嗜水气单胞菌主要外膜蛋白基因克隆及其表达

A pair of primers were designed according to the published nucleotide sequence of a putative outer membrane protein gene (omp) of Aeromonas hydrophila . With the specific primers, a target fragment about 1.1 kb was amplified from Aeromonas hydrophila ML316 via PCR .The target fragment was inserted into the linearized pGEM-T easy vector. After enzyme restriction and sequencing analysis,the nucleotide data had been further analyzed by DNAman and ClutalW software. The analysis results showed that the cloned DNA fragment had a longest open reading frame (ORF) of 1035 nt,it predicted to be encoded a 344 aa protein with the molecular weight of 36 kD. Hydrophobicity analysis suggested that the protein was highly hydrophilic, especialy at the first 24 aminoacid, this region could function as signal peptide. The homologious comparison proved the cloned gene had 96% homology to the sequence of the omp gene, and the alignment of the amino acid sequence was 98% . The recombinant plasmid was constructed with the target gene and the expressing vector pGEX-4T-1 and then was transformed into E. coli BL21（DE3）by BamH and Sal I .The fusion protein was expressed under the IPTG inducing condition,and exhibited about 62 kD in size,very close to the predicted molecular weight of GSTMOMP, furthermore,the fusion protein was specifically recognized by antiserum which raised against the major outer membrane protein of AHML316. Considering all these together, it proved that the cloned gene represented the major outer membrane protein gene of AHML316, and the expressed gene products shared identical antigenicity with the natural main outer membrane protein,and also provided technical support for developing an advanced gene engineering vaccine against Aeromonas hydrophila.     

大菱鲆仔鱼皮肤发育及超微结构

The histological development and ultrastructure of larval turbot skin were studied by both light and transmission electron microscopes.The early larvae showed a complete bisymmetry of skin.The skin of 1-25days old larvae developed slowly and was composed of the thin epidermis of 1-2layers epidermal cells and undeveloped dermis before the metamorphosis.But the skin developed significantly when the.metamorphosis began.Till the completion of metamorphosis(about 60days old),the skin contained 3-4layers epidermal cells and very developed collagenous strata.Wisth the process of metamorphosis,the ocular side skin and blind side skin became different in ultrastructure.Ultrastructural observation showed the epidermis of turbot contained three types of epidermal cells;filament-containing cells,mucus cells and chloride cells.The ocular side epidermis with looser structure consisted one type of filament-containing cells,but the blind side epidermis was structured densely and two types of filament-containing cells were observed in it.Melanophores,iridophores,fibroblasts and other cell types and tisssues were found in the spongiosum of dermis.The distribution of melanophores depended on the larval developing stage and the pigmentation of skin.The differentiation of skin seems to be an adaptation for the change of life style of turbot larvae from pelagic living to benthonic living after the metamorphosis.     

羊栖菜和裙带菜中抗凝血活性物质的初步筛选

Components with blood-anticoagulant activity were fractionated from marine algae Sargassumfusiforrne and Undaria pinnatifida and the activity of the components was investigated by using the method of bioassay. It was found that the 80% ethanol soluble fraction of Sargassumfusiforme exhibited no blood-anticoagulant activity, while the components obtained from the 80 % ethanol insoluble fraction (hot water extract) gave obvious bloodanticoagulant activity. A further study suggests that the principal component in the hot water extract from Sargassumfusiforrne with high blood-anticoagulant activity should be sulfated polysaccharides, in which the activity was positively correlated with the content of total sugar and fucose in sulfated polysaccharides. As for Undaria pinnatifida, the n-butanol extract fractionated from the 80% ethanol soluble fraction had blood anticoagulant activity, while the petroleum ether, ether, ethyl acetate, or water extract showed no bloodanticoagulant activity.     

^60Co-γ射线对成体长牡蛎生长的影响及致死效应

Effects of ionizing irradiation of ^60Co-γ ray on mortality and growth of one year-old Pacific oysters ( Crassotrea gigas ) were studied. Irradiation was conducted at 4℃ and without sea water, and the survival, shell length, shell width, whole wet weight and condition index of irradiated oysters were measured. A biphasic dose response effect of 6^60Co-γ irradiation on Pacific oyster with high stimulatory response dose range to 100 Gy, and strong hormesis effects had been observed. Gamma ray induced mortality happened within 90 days after irradiation, and the relation between dosage ( D ) and mortality at 90 days could be described by the equation of M90 = 0.0041D^2- 0. 1816D (R^2= 0. 9958) with an LD50/90 of 135 Gy, indicating that the Pacific oyster has strong resistance to ^60Co-γ irradiation. Growth in shell length, shell width and whole wet weight of Pacific oyster were also enhanced by 60 Gy of ^60Co-γ ray. The condition index was also enhanced by ^60Co-γ irradiation in the dosage range from 40 to 100 Gy, with the maximum stimulatory response of 2.5 fold of control value at the 40 Gy dosage, suggesting that the accelerating effects of γ ray on growth of the Pacific oyster led to increase in dry meat. Results of this study showed that ionizing irradiation might be useful in the improvement of production traits in shellfish.     

添加外源益生菌对大菱鲆育苗生物饵料菌群结构的影响

为评价在大菱鲆育苗生产中添加外源益生菌对生物饵料轮虫和卤虫微生物菌群结构的影响,运用基于illumina HiSeq平台的高通量测序技术对添加益生菌和按照生产流程正常强化的轮虫、卤虫进行了菌群结构的分析和对比。添加外源益生菌的实验组轮虫和卤虫中的菌群物种多样性均明显高于对照组。在轮虫强化过程中,不同时期的对照组样品中菌群结构差异较明显,优势细菌种类变化较大。而添加外源益生菌后的各时期实验组轮虫菌群结构很相似,优势菌群的种类更为丰富,Lactococcus sp.、Pseudoalteromonas sp.和Alteromonas sp.等一直是各实验组中的优势细菌。在卤虫强化过程中,各对照组样品的菌群结构高度相似,优势细菌Cobetia sp.的相对丰度高达54%~65.2%。而在添加益生菌后,各实验组中的菌群结构仍高度相似,但Cobetia sp.的比例下降至4.3%~25.3%,最优势的细菌为Pseudoalteromonas sp.和Alteromonas sp.等几种,菌群结构中的物种均匀度更好。研究表明,在轮虫和卤虫强化过程中添加外源益生菌,能够改变生物饵料的菌群结构,使生物饵料中的细菌种类均匀度更好,并使菌群结构趋于稳定。     

Microbially matured water: a technique for selection of a non-opportunistic bacterial flora in water that may improve performance of marine larvae

Before transfer to larval incubators, water was membrane filtered to remove >95% of the bacteria and then transiently maintained in a biofilter that promoted recolonization of the water by non-opportunistic bacteria. The process is termed microbial maturation of the water. Hypothetically the bacterial flora in the matured water should protect the marine larvae from colonization and proliferation by opportunistic bacteria. Testing of the hypothesis demonstrated 76% higher survival of yolk sac larvae of Atlantic halibut (Hippoglossus hippoglossus) in matured than in membrane filtered water. Proliferation of opportunistic bacteria was observed in the rearing water after hatching of turbot eggs (Scophthalmus maximus), but to a less extent in the microbially matured water. In the early phase of first feeding of turbot larvae, the matured water induced qualitative differences in the gut microflora. Significantly higher initial growth rate of the turbot larvae in the matured water affected 51% higher average weight of 13 days old larvae than in membrane filtered water. Algal addition to the matured water enhanced the larval growth further. The experiments conducted supported the proposed hypothesis that microbial maturation selects for non-opportunistic bacteria, which protects the marine larvae from proliferation of detrimental opportunistic bacteria.     

大菱鲆仔稚鱼发育早期肠道菌群结构形成的影响因素

为研究鱼类发育早期肠道菌群结构的演变过程及影响因素,运用高通量测序技术,分析了处于不同发育阶段的大菱鲆(Scophthalmus maximus)仔稚鱼肠道、受精卵、不同类型的饵料和水源中的菌群结构,以及它们之间的相关性。结果显示,以不同的OTU(Operational Taxonomic Units,可操作分类单元)作为分类依据,发现大菱鲆仔稚鱼的肠道菌群结构在开口摄食后不久已趋于稳定,其优势菌与受精卵所携带的细菌关联较大。并且在大菱鲆仔稚鱼不同的发育时期,这一菌群的结构非常稳固,几乎不受水和饵料中优势细菌的影响而发生改变。乳球菌属的Lactococcus piscium菌株一直是大菱鲆仔稚鱼肠道中的优势菌种,在不同发育时期的优势度高达45%~65%。本研究还发现,大菱鲆仔稚鱼肠道可能对定植的菌种具有选择性,一些水环境和饵料中的非优势菌,如Streptococcus sp.,Pseudomonas sp.,Carnobacterium sp.等细菌也会定植于肠道,成为大菱鲆肠道中的次优势菌。     

中华绒螯蟹温室育苗的水处理

通过对生物净化系统处理前后育苗水质指标变化情况的测定,初步探讨了生物净化系统在河蟹育苗过程中对水质的处理效果和应用价值。与传统的育苗工艺相比,应用生物净化系统有机物降解速度快,脱氮效果显著。ＣＯＤ、总氨氮的平均去除率分别为３３．２％,５５．５％。整个育苗过程ｐＨ稳定,溶氧丰富,池底无氧债和黑臭。     

温度对大菱鲆幼鱼生长及免疫相关酶活性的影响

将体质量(31.13±1.67)g的大菱鲆幼鱼饲养在容积100L的水桶中,每个水桶40尾,采用OKE-6710HF微电脑温控仪控制温度为15、18、21、24℃和27℃,在第0、15、30、45、60d进行采样分析血清中免疫相关酶活性及幼鱼全长、体质量及存活率。结果显示,在15~27℃条件下,大菱鲆幼鱼生长速度随温度升高先升后降;水温为18℃时,幼鱼存活率为100%,生长速度最快,为最佳生长水温。大菱鲆幼鱼血清中酸性磷酸酶和碱性磷酸酶活力,随温度及饲养时间的变化而呈波动性,但没有表现出显著的规律性;大菱鲆幼鱼血清中超氧化物歧化酶的活力随温度的升高先升后降;刚达到设定温度(即第0d)时,各温度组之间溶菌酶活力并无显著差异(P0.05),随着饲养时间的加长及温度的升高,溶菌酶活力不断升高,表明温度对大菱鲆幼鱼生长及存活率具有显著的影响。     

鳜不同养殖模式水质因子变化规律和微生物制剂调节技术研究

该文采用生态学试验方法,对鳜池塘和大棚养殖模式的水质变化规律进行了调查分析,同时采用不同微生态制剂商品(光合细菌、枯草芽孢杆菌、乳酸菌)对鳜养殖水体水质调节效果进行了研究。结果显示,整个鳜养殖周期(苗种至商品鱼),大棚养殖模式水体温度、溶氧、pH值与池塘养殖模式无明显区别;大棚养殖模式三氮(铵态氮、硝态氮、亚硝态氮)变化规律与池塘养殖模式大致相同,但大部分时间前者水体含量较高;大棚养殖模式水体总磷含量高于池塘养殖模式,且总磷最高值出现时间较池塘养殖模式推迟了近1个月。光合细菌对鳜养殖水体氨氮、亚硝态氮以及总氮整体调控效果最佳;枯草芽孢杆菌对降低硝态氮和亚硝态氮有良好的效果;乳酸菌对养殖后期降低水体pH值有一定的作用。结论:相对于鳜池塘养殖模式,大棚养殖模式氮磷物质循环转化效率较低,合理搭配使用微生物制剂调节水质养殖效果更佳,同时需注意不良天气对微生态制剂使用效果的影响。     

The effect of ammonia on the growth of juvenile Dover sole, Solea solea (L.) and turbot, Scophthalmus maximus (L.)

The growth of sole and turbot was followed in groups of fish exposed to constant unionised ammonia concentrations at pH levels both at and below that of normal sea water. Low pH levels were produced by the addition of carbon dioxide. Results for both species showed evidence for a threshold level of unionised ammonia below which little or no effect on growth was evident. This level was determined to be 0.066 mg N/l for sole and 0.11 mg N/l for turbot at 16°C, 34‰ salinity. The different pH conditions did not have any marked effect on these threshold levels.Above the threshold levels growth appeared to be depressed in a linear manner with increased unionised ammonia. The rate at which growth was depressed was greatest at the low pH levels for both species. The level at which growth ceased thus decreased from 0.77 to 0.38 mg N/l for sole and from 0.9 to 0.3 mg N/l for turbot, over the pH range from 7.9 to 6.9.The absence of disease and gill damage in the experimental populations is discussed. It is suggested that this and the tolerance shown by the fish, which is higher than is generally accepted for fish in high density culture, may be due to the clean environment in which the fish were held while under test.     

Colonization of the gut in first feeding turbot by bacterial strains added to the water or bioencapsulated in rotifers

Two bacterial strains, 4:44 and PB52, isolated from turbot(Scophthalmus maximus L.) were used during a first feedingexperiment with turbot larvae. Bacteria were either added directly tothe water on the day of hatching of the larvae (day 0), orbioencapsulated in rotifers (Brachionus plicatilis) distributedon day 2 after hatching. The two bacterial strains were found to bepresent in the water of the rearing tanks throughout the experiment. Theaddition of bacteria influenced the species-composition of themicroflora associated with intensively produced rotifers added to thetanks, and resulted in colonization of the gut of the larvae by theadded bacterial strains. The strain 4:44 showed a peak on day 9 afterhatching, and reached a mean(sem) of 2.5(1.4) × 10⁴bacteria per larva, when added in a mixture with PB52, and 0.4(0.1)× 10⁴ bacteria per larva, when added alone. The strainPB52, on day 12 after hatching, reached 5.2(1.5) × 10⁴bacteria per larva when added in a mixture with strain 4:44, and12.5(0.7) × 10⁴ bacteria per larva, when added alone.The added bacteria colonized the gut turbot larvae, whereas thebacterial land, the survival and growth of the larvae were in most casesnot influenced in a negative way by the addition of bacteria.     

三种微生物制剂对有机物废水中氨氮及亚硝酸盐的影响

实验采用复合净水菌、复合芽孢杆菌、光合细菌与空白对照比较了三种微生物制剂对有机物废水中氨氮及亚硝酸盐的处理效果。结果显示：清除氨氮方面：利用微生态制剂微生物制剂净化7d,三种微生物制剂对有机物污染水体中氨氮的清除率均极显著大于对照组（P〈0.01）,处理组之间复合芽孢杆菌的氨氮降解率56.58%,显著大于降解率分别为42.64%和44.02%的复合净水菌和光合细菌组（0.01〈P〈0.05）。清除亚硝酸盐方面：三种微生物制剂对亚硝酸的降解率均极显著大于对照组（P〈0.01）,而复合芽孢杆菌对亚硝酸的降解率68.84%,显著大于复合净水菌47.70%及光合细菌组37.17%（0.01〈P〈0.05）。结果表明：三种微生物制剂对有机物污染的水体中的氨氮及亚硝酸盐均具有降解效果,其中复合芽孢杆菌对于降低水体中氨氮和亚硝酸盐效果最佳。     

硝化细菌制剂对淡水水族箱水质的净化效果

研究自制硝化细菌制剂对水族箱水质的净化效果,试验结果表明:硝化细菌制剂对水族箱水质具有明显的净化效果,实验组水族箱水质氨氮、亚硝氮、COD等指标明显低于对照组。投加硝化细菌制剂后,水族箱内氨氧化细菌、亚硝酸盐氧化细菌可在短时间内形成优势,使氨氮、亚硝氮维持在较低浓度水平;在不投加菌剂的情况下,氨氧化细菌虽然可在一定时间内形成优势,使氨氮浓度降低,但由于亚硝酸氧化细菌生长缓慢,水族箱中亚硝酸积累问题严重。