粪肠球菌生长曲线的测定及其对小鼠脑组织的影响

为测定致羔羊脑炎粪肠球菌（Enterococcus faecalis）的生长曲线，寻求一种快速而准确的方法测定不同生长时期粪肠球菌数量，并客观评价其毒性强弱及其对小鼠脑组织的影响，试验采用平板菌落计数法和OD-Monitor振荡比浊法（D_λ值法）测定粪肠球菌的生长曲线，探究该菌在合适时间段内的吸光值（D_{600 nm}）与平板菌落计数法测定的活菌数（CFU）的关系。用粪肠球菌感染小鼠，观察记录小鼠的死亡情况，最后采用Karber法计算粪肠球菌感染小鼠的半数致死量（LD₅₀）。用LD₅₀的剂量感染小鼠，及时采集死亡小鼠脑组织，未死亡的小鼠72 h后全部剖杀取脑组织，一部分做涂片染色，制作病理切片，观察病理变化；一部分进行培养，用于PCR方法进行细菌的回收鉴定。结果显示，用两种方法测定此株粪肠球菌的生长曲线基本一致，在2~8 h生长迅速，为对数生长期，8~14 h生长缓慢，为稳定期，14 h之后死亡数增加，进入衰亡期；对12 h粪肠球菌D_{600 nm}与CFU的关系进行探讨，成功建立回归方程：y=20.769x-1.3422，R²=0.997；其感染小鼠的LD₅₀为7.77×10¹¹个活菌。以此剂量感染小鼠，脑组织涂片染色和培养染色，均能看到革兰氏阳性球菌；PCR结果显示，均出现了大小为112 bp的条带。对脑组织进行病理学观察发现该菌可导致脑组织充血、出血、形成微血栓，脑膜充血。通过生长曲线和其D_{600 nm}与CFU关系的建立，可实时监测粪肠球菌数量，为后期更深入研究粪肠球菌穿越血脑屏障的机制奠定重要的理论基础。     

水流作用下一种养殖金属网衣水阻力特性的数值模拟研究

为保障深远海金属网箱养殖系统的结构安全,进行网箱结构的水动力学研究是一项重要的工作。构成金属网箱主要组成部分的金属网衣是一种小直径多孔的网状结构,在波浪和水流作用下的水动力学特性与一般的海洋工程结构物有显著不同。本文基于有限元基本原理,采用梁单元模拟金属网线结构,采用连接单元模拟网线接触部位的相互作用,并运用ABAQUS软件进行数值计算,在试验验证的基础上,分析了在水流作用下一种金属菱形链网衣在不同网目尺寸、不同网线直径情况下的水阻力变化情况。数值模拟结果表明在各种工况下,当网目尺寸由25 mm增加到35 mm、45 mm时,网衣受力增加幅度的平均值分别为14.71%和38.07%;当网线直径由2.5 mm增加到3.2 mm、4.0 mm时,各工况下网衣受力增加幅度的平均值分别为25.05%和45.06%。研究结果可以为进一步开展深海养殖金属网箱水动力特性研究提供积极的基础。     

闽江流域表层沉积物硫化物及重金属生物毒性研究

根据2012—2013年福建省闽江流域富屯溪、建溪和沙溪等支流以及干流7个站位表层沉积物调查数据,对硫化物(AVS)、有机碳及重金属(SEM)等物质进行了研究。结果表明,各站位硫化物两年平均含量在0.461 0~1.814 0μmol·g~(-1)之间,有机碳平均含量在0.76%~1.51%之间,尤溪S5站位硫化物含量明显高于其他站位,与此对应有机碳含量也较高,这与沿岸人类活动的强度有关;闽江干流S7站位硫化物含量最低,主要受水动力特征影响。各站位沉积物重金属(Cu、Pb、Cd、Hg和As)含量基本呈现CuPbAsHgCd的趋势,其中Cu和Pb是主要污染重金属。SEM平均含量在0.47~3.58μmol·g~(-1)之间,金溪S2、古田溪S6站位SEM/AVS(摩尔比值)小于1,重金属潜在生态危害风险较小;其他站位SEM/AVS大于1,生物有效性高,具有潜在的毒性效应。与水体沉积物中金属元素对底栖生物产生毒性效应的阈值比较,单个重金属中Pb和Cd可能产生的生物有效性高于Cu,具有对底栖生物产生毒性效应的潜力。     

Polymerisation of gluten proteins in developing wheat grain as affected by desiccation

The breadmaking quality of wheat is affected by the composition of gluten proteins and the polymerisation of subunits that are synthesised and accumulated in developing wheat grain. The biological mechanisms and time course of these events during grain development are documented, but not widely confirmed. Therefore, the aim of this study was to monitor the accumulation of gluten protein subunits and the size distribution of protein aggregates during grain development. The effect of desiccation on the polymerisation of gluten proteins and the functional properties of gluten were also studied. The results showed that the size of glutenin polymers remained consistently low until yellow ripeness (YR), while it increased during grain desiccation after YR. Hence, this polymerisation process was presumed to be initiated by desiccation. A similar polymerisation event was also observed when premature grains were dried artificially. The composition of gluten proteins, the ratios of glutenin to gliadin and high molecular weight-glutenin subunits to low molecular weight-glutenin subunits, in premature grain after artificial desiccation showed close association with the size of glutenin polymers in artificially dried grain. Functional properties of gluten in these samples were also associated with polymer size after artificial desiccation.     

Disulphide structure of high-molecular-weight (HMW-) gliadins as affected by terminators

This study aimed at elucidating SS-bonds of HMW-gliadins (HGL) from wheat with the focus on terminators of glutenin polymerisation. HGL from wheat flour extracts non-treated or treated with the S-alkylation reagent N-ethylmaleinimide (NEMI) were compared. HGL from wheat flour Akteur were isolated, hydrolysed with thermolysin and the resulting peptides pre-separated by gel permeation chromatography and analysed by liquid chromatography/mass-spectrometry using alternating electron transfer dissociation/collision-induced dissociation. Altogether, 22 and 28 SS-peptides from samples without and with NEMI treatment, respectively, were identified. Twenty-six peptides included standard SS-bonds of α- and γ-gliadins, high-molecular-weight and low-molecular-weight glutenin subunits. Eleven SS-bonds were identified for the first time. Fifteen peptides unique to HGL contained cysteine residues from gliadins with an odd number of cysteines (ω5-, α- and γ-gliadins). Thus, gliadins with an odd number of cysteines, glutathione and cysteine had acted as terminators of glutenin polymerisation. Decisive differences between samples without and with NEMI treatment were not obvious showing that the termination of polymerisation was already completed in the flour. The two HGL samples, however, were different in the majority of ten peptides that included disulphide-linked low-molecular-weight (LMW) thiols such as glutathione and cysteine with the former being enriched in the non-treated HGL-sample.     

The release and uptake of metals from potential biofilm inhibition products during spiny lobster (Sagmariasus verreauxi,H. Milne Edwards 1851) culture

Zinc (Zn) and copper (Cu) are strong inhibitors of bacterial biofilms in aqueous solutions, but are known toxins of crustaceans. A new metal application method; cold‐sprayed metal embedment, known to modulate metal release, was tested for its applications in crustacean larval culture systems. Cold‐spray technology allows metal particles to bond to plastics, while modulating metal ion release and biocide activity to the substrate boundary. In this study, Eastern spiny lobster (Sagmariasus verreauxi) larvae (phyllosoma) were cultured in the presence of cold‐sprayed Zn and Cu metal surfaces. Metal loss was monitored gravimetrically on embedded surfaces, assessment of water ion concentrations and analysis of phyllosoma body content were undertaken. Phyllosoma moulting, deformity and mortality patterns were monitored. Cold‐sprayed Zn‐ and Cu‐embedded surfaces were depleted with losses of 0.69% and 31.2% noted respectively. Culture water concentrations of these metals were elevated and accumulation by phyllosoma occurred. Water Zn concentrations of 18.5 μg L^?1 were associated with chronic eyestalk moult deformities; the first report of Zn causing a non‐lethal moult deformity in crustacean larvae. The Cu surface lost a third of its metal mass with a water concentration of 40 μg L^?1 causing acute toxicity and localization of composite granules in the midgut gland. Cu associated mortality was noted by Day 2 of culture with a LD 50 experienced by Day 9. Future work on the use of bioactive metals in aquaculture systems will focus on a range of different metal alloys, and improved modulation of ion release mechanisms through increased particle embedment depth and separation.     

Comparative functional responses of native and high‐impacting invasive fishes: impact predictions for native prey populations

Comparative functional responses (FRs) can predict impacts of invasive species, including piscivorous fishes, via quantifying their depletion of native food resources as a function of prey density. The utility of FRs for predicting impacts on prey populations by invasive fishes of different trophic guilds was tested here by comparing the FRs of the invaders Cyprinus carpio and Carassius auratus, with three native, trophically analogous fishes, Barbus barbus, Squalius cephalus and Tinca tinca. Chironomid larvae and Gammarus pulex were used as prey items. Predictions, developed from studies on the foraging of C. carpio and C. auratus in the literature, were that the invaders would have significantly higher consumption rates for chironomids than the native fishes, but not for G. pulex. Mean consumption rates for chironomids were significantly lower for both invaders than B. barbus and S. cephalus, but were similar to T. tinca. Barbus barbus had a significantly lower consumption rate of G. pulex than both invaders, but there were no significant differences between S. cephalus, T. tinca and the invaders. All FRs were type II, with FR curves for the invaders preying upon chironomids never being significantly higher than the native fishes, contrary to predictions. For G. pulex, some significant differences were apparent between the invaders and native fishes, but again were contrary to predictions. These results indicated that when predation impacts of invasive fishes could also be a function of their population density and body sizes, these parameters should be incorporated into FR models to improve impact predictions.     

Long‐term habitat loss in a lightly‐disturbed population of the Indo‐Pacific humpback dolphin,Sousa chinensis

1. Coastal and estuarine waters are important ecosystems with high primary and secondary productivity, but they are prone to the impacts of habitat loss caused by anthropogenic activities. For species exclusively inhabiting coastal and estuarine waters, such as the Indo‐Pacific humpback dolphin, Sousa chinensis , irreversible habitat loss can have dramatic implications for population viability.
2. A Landsat image database was used to determine the extent of coastal changes along the northern Beibu Gulf, where a large humpback dolphin population is found. The results were compared with the standardized sighting gradient (SPUF) determined from a questionnaire survey of fishermen and likely core habitats identified by application of a global digital elevation model.
3. Both SPUF and likely core habitat results indicated a continuous distribution of the humpback dolphin along the northern Beibu Gulf. Landsat images revealed that 129.6 km² of coastal waters were permanently lost in the past 40 years, 60 km² within the likely core habitats. Although this may be considered small, the impact of such habitat loss could be substantial in some local habitats.
4. The humpback dolphin population in the northern Beibu Gulf should be regarded as one management unit, with two or more social subunits. Immediate systematic surveys are needed to fill information gaps on true distribution range and habitat‐use patterns.
5. Habitat protection actions for dolphins in the northern Beibu Gulf should include both core and linking habitats, including enacting protected areas in core habitats, mitigating anthropogenic impacts in likely habitats, restoring both coastal waters and surrounding landscape quality, effective treatment of industrial sewage discharge, and comprehensive environmental impact assessments for the planning of coastal development projects.

     

Replacement of Fishmeal by a Mixture of Soybean Meal and Chlorella Meal in Practical Diets for Juvenile Crucian Carp,Carassius auratus

A growth trial was conducted to evaluate the effect of a mixture of soybean meal and Chlorella meal (SCM) as a dietary fishmeal (FM) substitute on growth performance, apparent digestibility coefficients (ADCs), digestive enzymatic activities, and histology of juvenile crucian carp, Carassius auratus. Five isonitrogenous and isoenergetic diets were formulated to replace 0 (SCM0), 25 (SCM25), 50 (SCM50), 75 (SCM75), and 100% (SCM100) of protein from FM with SCM, respectively. The diets were fed to triplicate groups of juvenile crucian carp for 6 wk. Weight gain, specific growth rate, feed intake, protein efficiency ratio, and intestinal digestive enzymatic activities (amylase, trypsin, and lipase) tended to decline with increasing FM replacement levels (P > 0.05). Dietary SCM substitution significantly influenced dry matter content in muscle, and crude protein and lipid contents in liver (P < 0.05). ADCs for dry matter, protein, lipid, energy, and most amino acids showed no significant differences between the control and SCM25 group, but tended to decline with replacement levels over 25%. Higher SCM substitution (50–100%) caused karyopyknosis and necrosis in liver, but intestinal histology did not show noticeable pathological changes. The present study indicated that FM could be replaced by 25% of SCM, without significant adverse growth performance, feed utilization, and histology of crucian carp.