The aniline blue fluorochrome specifically stains the septum of both live and fixed Schizosaccharomyces pombe cells

Abstract A novel method is described which uses aniline blue for the specific fluorescent staining of the septa of dividing cells of the fission yeast, Schizosaccharomvces pombe . It gives the same results with live and fixed cells. In fixed or, more generally, dead cells there is no staining of the cytoplasm: this renders aniline blue superior to other dyes previously used to stain the septum of S. pombe . This feature allows quantitative analysis of the septum index for fixed samples and, therefore, makes aniline blue the stain of choice for cell cycle kinetic studies.     

伊红、台盼蓝检测河蟹精子存活率的比较

对台盼蓝和伊红染色法检测河蟹(Eriocheir sinensis)精子存活率的方法进行了评价研究。结果表明,两种染色法死、活精子分别呈现出明显不同的染色特征:活精子无色透明,顶体中央凸起呈圆锥状,光镜下辐射臂及细胞边界清晰;死亡精子顶体着色,且中央有一染色较深的圆斑,核杯染色不明显,细胞体积变大,边界模糊。通过不同染色时间和不同染料浓度的比较发现,两种染色法最适染液浓度分别是0·25%的伊红和0·5%的台盼蓝,染色时间均以15min为佳。在此基础上,将新鲜精子和60℃水浴处理致死精子以不同的体积比混合,配成含致死精子比例为10%～90%的9个梯度样品,用伊红和台盼蓝分别测定各样品精子死亡率,并进行相关性分析。结果发现,各样品实测精子死亡率均略高于样品的理论死亡率,同时两种染色法实测值与样品理论值呈显著正相关(P<0·05),两种染色法之间亦呈显著正相关(P<0·05)。上述结果表明,伊红和台盼蓝可用于河蟹精子的活体染色,且两种染色法在对河蟹精子染色中具有一定的稳定性和可比性。     

Excretion of heptadecene-1 into lake water by swarms of Polyphemus pediculus (Crustacea)

1. In the littoral zone of a mesotrophic freshwater lake (Lake Selent, Northern Germany) much higher concentrations of heptadecene-1 were found within a swarm of Polyphemus pediculus than in lake water at some distance from the swarm. Polyphemus collected in the field produced large amounts of heptadecene-1 when exposed to osmotic shock. Other volatile compounds were not detected.
2. Heptadecene-1 could be involved in the formation, maintenance and recognition of swarms of Polyphemus. It is the first report of the release of a volatile organic compound from a freshwater crustacean.     

The effect of small zooplankton on the microbial loop and edible algae during a cyanobacterial bloom

SUMMARY 1. We studied the effect of the small crustacean zooplankton on heterotrophic micro-organisms and edible phytoplankton in a eutrophic lake during a cyanobacterial bloom.
2. Small (15 L) enclosures were filled with natural or screened (100 μm) lake water and incubated for 5 days in the lake. Screening removed crustacean zooplankton but the initial density of rotifers and phytoplankton remained the same in control and removal treatments. Changes in the abundance and biomass of bacteria, autotrophic picoplankton (APP), heterotrophic nanoflagellates (HNF) and ciliates were measured daily.
3. The crustacean zooplankton, dominated by the small cladoceran Chydorus sphaericus , did not affect cyanobacteria, the main phytoplankton group during the experiment.
4. The removal of the crustacean zooplankton induced a higher abundance of ciliates and reduced that of the HNF, indicating the importance of ciliates in controlling HNF in this system.     

Temporal and spatial patterns of crustacean zooplankton dynamics in a transitional lagoon ecosystem

Patterns and mechanisms of plankton crustacean seasonal succession in the eutrophic freshwater Curonian lagoon (south-eastern Baltic Sea) were analysed on the basis of four-year (1995, 1996, 1998 and 1999) field sampling results. The seasonal crustacean zooplankton succession in the lagoon appears to be the consistent six-stage sequence of four distinct species complexes. Each stage is characterised by its individual species composition and quantitative characteristics. The uniform and periodic pattern of the limnetic zooplankton crustacean successional stages in the lagoon indicates that the seasonal succession of the limnetic zooplankton is not disturbed by unpredictable environmental fluctuations, such as brackish water inflows. Seasonal zooplankton succession is also comparatively uniform at a spatial scale. Not more than two adjacent successional stages were found across the northern part of the lagoon during each of 11 seasonal surveys. Comparison between monthly water residence time and dominant plankton crustacean species life cycle duration points to a more transitory plankton community in spring while in the summer it is not much influenced by lagoon hydrodynamics. Consequently, the Curonian lagoon crustacean community quite closely follows the Plankton Ecology Group (PEG)-described freshwater lake seasonal succession in summer and turns into a lentic-like system in spring and autumn.     

Changes in the crustacean zooplankton community of Harp Lake, Canada, following invasion by Bythotrephes cederstrœmi

1. Detecting the impacts of invading Bythotrephes cederstrœmi (Crustacea, Onychopoda, Cercopagididae) on zooplankton in North American lakes has been hampered by the brevity of pre-invasion data, and by the difficulty of distinguishing the effects of the invader from other stresses. The data from Harp Lake in Ontario, Canada, circumvent these difficulties. Bythotrephes appeared in the lake in 1993. There is a 15-year pre-invasion data set, and no significant complicating concurrent stresses.
2. The species composition and the size structure of the crustacean zooplankton community of Harp Lake changed after the invasion. Several small species either declined dramatically in abundance (e.g. Bosmina longirostris , Tropocyclops extensus ) or disappeared ( Chydorus sphaericus , Diaphanosoma birgei , Bosmina ( Neobosmina ) tubicen ). In contrast the abundance of the larger cladocerans Holopedium gibberum and Daphnia galeata mendotae and the hypolimnetic copepod Leptodiaptomus sicilis increased. Several univariate and all multivariate summarizations of zooplankton abundance, biomass and size structure highlighted the uniqueness of the post-invasion community.
3. The alterations in the zooplankton community could not be attributed to changes in lake acidity, thermal regimes, penetration by ultraviolet light, nutrient status, fish stocking or the abundances of native invertebrate predators, but they were correlated with Bythotrephes abundance, both within and among years. Hence, we hypothesize that the invasion by Bythotrephes has significantly altered the crustacean zooplankton community of Harp Lake.     

Changes in the crustacean zooplankton community of Harp Lake, Canada, following invasion by Bythotrephes cederstrœmi

1. Detecting the impacts of invading Bythotrephes cederstrœmi (Crustacea, Onychopoda, Cercopagididae) on zooplankton in North American lakes has been hampered by the brevity of pre-invasion data, and by the difficulty of distinguishing the effects of the invader from other stresses. The data from Harp Lake in Ontario, Canada, circumvent these difficulties. Bythotrephes appeared in the lake in 1993. There is a 15-year pre-invasion data set, and no significant complicating concurrent stresses.
2. The species composition and the size structure of the crustacean zooplankton community of Harp Lake changed after the invasion. Several small species either declined dramatically in abundance (e.g. Bosmina longirostris , Tropocyclops extensus ) or disappeared ( Chydorus sphaericus , Diaphanosoma birgei , Bosmina ( Neobosmina ) tubicen ). In contrast the abundance of the larger cladocerans Holopedium gibberum and Daphnia galeata mendotae and the hypolimnetic copepod Leptodiaptomus sicilis increased. Several univariate and all multivariate summarizations of zooplankton abundance, biomass and size structure highlighted the uniqueness of the post-invasion community.
3. The alterations in the zooplankton community could not be attributed to changes in lake acidity, thermal regimes, penetration by ultraviolet light, nutrient status, fish stocking or the abundances of native invertebrate predators, but they were correlated with Bythotrephes abundance, both within and among years. Hence, we hypothesize that the invasion by Bythotrephes has significantly altered the crustacean zooplankton community of Harp Lake.     

Dead cell counts during serum cultivation are underestimated by the fluorescent live/dead assay

The live/dead fluorescent assay provides a quick method for assessing the proportion of live and dead cells in cell culture systems or tissues and is widely used. Dead cells are detected by the fluorescence produced when propidium iodide (PI) binds to DNA; PI and similar molecules are excluded from live cells but can penetrate dead cells because of their loss of membrane integrity. Here we investigated the effect of serum in the culture medium on the reliability of the method. We assessed viability of chondrocytes with/without serum using both a live/dead assay kit and also trypan blue staining. We found that after 2 days of culture, the DNA-binding dye PI could no longer detect dead cells if serum was present but they were readily detected in serum-free medium or if an inhibitor to DNase I was added to the serum-containing medium. Dead cells could be detected by trypan blue staining in all cultures. Hence dead cells are no longer detected as the DNase I present in serum degrades their DNA. DNA-binding dyes may thus not give a reliable estimate of the number of dead cells in systems that have been cultured in the presence of serum for several days.     

新疆乌伦古湖浮游甲壳动物的季节演替及与环境因子的关系

通过2006年11月至2008年8月2周年8个季度的野外采样调查和室内分析,研究了新疆乌伦古湖浮游甲壳动物的季节演替格局及其与环境因子的相关性。结果表明,枝角类和桡足类的总种类数为25种,其中枝角类6种,桡足类19种。枝角类Shannon-Wiener指数2007年夏季和2008年夏季较高,2007年冬季最低;Pielou 指数2006年冬季最高;桡足类Shannon-wiener指数和Pielou指数除冬季较低外,其余三季较高。方差分析结果表明枝角类和无节幼体的密度与生物量呈现出显著的季节变动格局,其中,枝角类密度和生物量在夏秋季形成单一高峰,最大值出现在2007年夏,最大密度和生物量分别为2.12个/L和0.062 mg/L,而无节幼体密度和生物量形成春秋双高峰,其最大值出现在2007年春,密度和生物量分别为5.14个/L和0.015 mg/L,桡足类密度和生物量季节变化不明显。优势种的季节演替方面,象鼻溞和模式有爪猛水蚤全年形成优势,其他枝角类、哲水蚤和剑水蚤不同季节的优势种各异。经典范对应分析(CCA)结果表明,长刺溞等与水体中的pH值、SD呈正相关,与浮游植物生物量呈负相关;长额象鼻溞等与水深呈正相关,无节幼体等与水温也呈正相关性。     

武汉东湖夏冬两季浮游动物物种多样性及群落结构研究

2018年7月、8月、12月及2019年1月, 对武汉东湖沿岸带的浮游动物群落分夏冬两季进行了调查研究。共鉴定出浮游动物35属50种, 其中轮虫22属37种、枝角类10属10种、桡足类3属3种。夏季优势物种为跨立小剑水蚤(Microcyclops varicans)、裂足臂尾轮虫(Brachionus diversicornis)和微型裸腹溞(Moina micrura); 冬季优势物种为长肢多肢轮虫(Polyarthra dolichoptera)、疣毛轮虫(Synchaeta spp.)、角突臂尾轮虫(Brachionus angularis)和犀轮虫(Rhinoglena sp.)。将观察到的浮游动物进行了图像信息采集, 并确定了部分物种SSU rRNA序列, 为淡水浮游动物的研究提供了参考资料。参考武汉东湖浮游动物研究的历史文献数据, 东湖数十年来浮游动物物种数逐渐下降, 轮虫优势物种由寡污型转为β-中污型甚至α-中污型。夏冬两季东湖浮游动物群落结构具有明显差异, 夏季轮虫平均密度(56.2 ind./L)和生物量(0.22 mg/L)低于冬季(476.3 ind./L、0.44 mg/L), 而甲壳类密度(137.8 ind./L)和生物量(2.88 mg/L)高于冬季(53.1 ind./L、1.12 mg/L)。夏季浮游动物群落Margalef丰富度指数、Shannon-Wiener多样性指数和Simpson多样性指数均高于冬季, 夏冬两季Pielou均匀度指数差异不大, 水质评价表明东湖夏季(中污染)优于冬季(重污染)。夏冬两季温度、溶解氧、pH、盐度和ORP五种环境因子均存在极显著差异, RDA分析表明, 其中温度对浮游动物群落结构的影响最大。     

Comparison of methods used in zooplankton sampling and counting in the joint Russian-Finnish evaluation of the trophic state of Lake Ladoga

The results of a joint Russian-Finnish investigation on zooplankton in Lake Ladoga are presented, and a comparison is made between two sampling techniques, tube sampler and plankton net, and between two counting methods. The precision of subsampling and sampling is further discussed on the basis of zooplankton data gathered in Lake Saimaa, Finland. The comparison clearly indicates that a tube sampler is required for reliable sampling of small-sized animals, while a plankton net saves time and is a more economical sampler for large, rare or active animals. The comparison between the results obtained by Finnish and Russian workers, using different counting procedures, shows that the main groups of crustacean zooplankton are similarly counted and identified in the two laboratories.     

浮游甲壳类定量采样计数方法评价

１引言浮游生物是养殖鱼类特别是滤食性鱼类的主要饵料,同时又是评价水生态系统质量优劣的重要指标．在大水面增养殖研究中,常常通过水体中浮游生物的采样分析,计算水体生物生产力．而在池塘生态系统中则又常常借助于浮游生物的采样分析来判定养鱼水质,决定增施饵肥料...     

Zooplankton biomass and community structure in a Danube River floodplain system: effects of hydrology

1. Zooplankton density and biomass was examined in a Danube River floodplain section with highly variable hydrological dynamics. Temporal patterns were analysed to assess the effects of hydrological conditions on zooplankton community structure and the differential response of the two major zooplankton taxa, rotifers and crustaceans.
2. Calculated floodplain water age was used as an integrated parameter describing hydrological conditions and connectivity.
3. Total zooplankton biomass, crustacean biomass and crustacean species number were significantly positively related to water age. Rotifer biomass followed a hump-shaped relationship with water age, and rotifer species number decreased with increasing water age.
4. Rotifers dominated the community in periods of low to medium water ages. In periods of higher water ages the community was dominated by crustaceans.
5. We propose that the hydrological regime of floodplains is crucial for zooplankton biomass patterns and succession, through the alternation of washing-out effects, taxon-specific potential of reproduction and biological interactions. Flood events and high water levels reset the community to an early successional phase.     

Comparison of Plankton-Water Chemistry Relationships in Three Acid Stressed Lakes in Nova Scotia,Canada

A field program was conducted in 1980 and 1981 on three lakes in Kejimkujik National Park, Nova Scotia, to study plankton-water chemistry relationships in a region of potential acid precipitation stress. Among lake and between year comparisons were made. The three study sites represent two different types of lakes. Kejimkujik Lake (pH 4.8) and Pebbleloggitch Lake (pH 4.5) are both high-colored, dystrophic acid lakes. Beaverskin Lake is a clear, oligotrophic lake which is not as acidic (pH 5.4). The phytoplankton of Kejimkujik Lake was dominated by diatoms, while Pebbleloggitch Lake was dominated by chlorophytes and chrysophytes, and Beaverskin Lake was dominated by cyanophytes. Kejimkujik Lake had the highest total algal cell volume per liter, and Pebbleloggitch Lake the lowest. Rotifer populations composed the majority of the zooplankton communities, while the crustacean zooplankton were dominated by the acid-tolerant copepod Diaptomus minutus. Kejimkujik Lake had the lowest zooplankton biomass and Pebbleloggitch Lake the highest zooplankton biomass per m³. A pattern of highest algal density in Beaverskin Lake, but highest zooplankton density in Pebbleloggitch Lake emerged in both years. This is likely an effect of abundant, but unsuitable food (blue-green algae) in Beaverskin Lake and important detrital food resources in Pebbleloggitch Lake in both years. Multiple regression analysis of water chemistry variables with plankton species produced many significant effects, but failed to show clear patterns. Cause and effect relationships in aquatic ecosystems are poorly delineated by such techniques.     

太湖梅梁湾大型控藻围栏对浮游甲壳动物群落结构的影响

2005年对太湖梅梁湾大型鲢、鳙控藻围栏内外浮游甲壳动物群落结构的季节变化进行了监测.结果表明:围栏内外的环境因子、浮游植物生物量、浮游甲壳动物种类组成无显著差异.但鲢、鳙放养对浮游甲壳动物的生物量产生了较大的影响,围栏内浮游甲壳动物的总生物量和枝角类的生物量显著低于围栏外.总体上,枝角类各种类的生物量受鲢、鳙放养的影响程度大于桡足类的种类.太湖梅梁湾浮游甲壳动物的季节演替明显,大部分种类只是季节性出现.冬季和春季以溞(Daphnia sp.)和近邻剑水蚤(Cyclops vicnus)等大型种类为主,夏季和秋季以象鼻溞(Bosmina sp.)、角突网纹溞(Ceriodaphnia cornuta)和中华窄腹剑水蚤(Limnoithona sinensis)等小型种类为主.典范对应分析表明,透明度、温度和浮游植物的生物量是影响浮游甲壳动物季节变化的主要因素.     

Microcystins derived from lysing Microcystis cells do not cause negative effects on crustacean zooplankton in Lake Taihu, China

Microcystins (MCs) have a toxic effect on crustacean zooplankton in the laboratory, but there is little or no unequivocal evidence in the literature of their lethal effects on crustacean zooplankton in the field. We used the natural microcystins extracted from Microcystis spp. to test if they could cause any negative effects on crustacean zooplankton. We conducted three experiments in enclosures with water from Lake Taihu, China, and microcystins derived by extraction from Microcystis spp. collected from the lake when the species was in bloom conditions. Initial concentrations of extracellular microcystins (EMCs = MC-RR + MC-LR + MC-YR) ranged from 9.7 to 44.9 μg/L in treatments with microcystin addition. Microcystin concentrations sharply decreased on second day in all the three experiments. EMCs at the end of the experiments varied from only 2.7 to 14.2 % of the levels at the start of the experiments. The dominant species of crustacean zooplankton in the lake were Bosmina longirotris, Ceriodaphnia cornuta, Mesocyclops spp., Limnoithona sinensis, Sinocalanus dorrii and Schmackeria inopinus. ANOVA analysis showed that the density and biomass of cladoceran and copepod did not significantly differ between treatments with microcystin addition and controls. Our results indicate that microcystins derived from lysing Microcystis do not cause any negative effects on crustacean zooplankton.     

鄱阳湖浮游甲壳动物群落结构特征

鄱阳湖是中国第一大淡水湖,具有"丰水为湖,枯水为河"的独特特点。为探讨鄱阳湖浮游甲壳动物群落结构及其时空分布的特征,于2009年全年采集其不同季节、不同水位期的浮游甲壳动物样品进行定量分析。结果显示,鄱阳湖浮游甲壳动物群落结构总体与河流浮游甲壳群落具有相似性。无节幼体、象鼻溞、剑水蚤等河流优势类群在鄱阳湖浮游甲壳动物中占优势;而哲水蚤和溞属仅在低水位季节占优势,枝角类丰度仅在高温、高水位、流速缓的季节高过桡足类。丰水期浮游甲壳动物平均丰度和生物量远远高于枯水期,可达枯水期的50倍,差异极其显著(P0.01)。温度和水位变化引起的环境因子改变是导致鄱阳湖浮游甲壳动物发生季节演替的主要原因;而营养盐对浮游甲壳动物的影响并不显著。空间上浮游甲壳动物群落构成明显不同,年均丰度最高和最低的点均出现在河口地区。因此:对于换水周期短,水交换速率快的水体,应该充分考虑水文条件对生物的影响。     

Viability and acrosome staining of stallion spermatozoa by Chicago sky blue and Giemsa

A simple trypan blue-neutral red-Giemsa staining procedure for simultaneous evaluation of acrosome, sperm head, and tail membrane integrity and morphology has been used to evaluate equine spermatozoa. Some special characteristics and problems have arisen in evaluating stallion semen. One problem was the differentiation of intact vs. damaged sperm tails primarily in frozen and thawed samples. After freezing and thawing, a high percentage of spermatozoa with an unstained head and stained tail were observed. These cells are considered immotile. Therefore, unambiguous differentiation of intact vs. damaged sperm tail membrane is very important for evaluating semen quality. The aim of our study was to develop a method especially for stallion sperm to distinguish more accurately the different cell types. We compared Chicago sky blue 6B (CSB) to trypan blue (TB) for viability staining. CSB/Giemsa staining showed good repeatability and agreement with TB/Giemsa measurements. For densitometry analysis, individual digital images were taken from smears stained by CSB/Giemsa and by TB/Giemsa. A red-green-blue (RGB) histogram for each area of spermatozoa was drawn. Differences of means of RGB values of live vs. dead tails and separate live vs. dead heads from each photo were used to compare the two staining procedures. CSB produced similar live/dead sperm head differentiation and better tail differentiation. TB can be replaced by CSB and this results in more reliable evaluation. After staining with 0.16% CSB and 4 min fixation, 2-4 h Giemsa staining at 25-40° C is recommended for stallion semen.     

The consumption of rainbow trout (Salmo gairdneri Richardson) eggs by macroinvertebrates in the field

SUMMARY. 1. Perforated boxes containing live, dead or no eggs of the rainbow trout, Salmo gairdneri Richardson, were buried in a natural spawning redd in a stream in North Wales for the duration of the natural incubation period.
2. A significant difference ( P <0.001) was found between the mean number of invertebrates colonizing perforated boxes containing live eggs, dead eggs and no eggs.
3. The number of eggs retrieved from boxes originally containing dead eggs decreased as the number of invertebrates colonizing the boxes increased.
4. Using a serological method of prey identification it was demonstrated that Leuctra hippopus (Kempny), Gammarus pulex (L.) and Ptychopter-idae sp. scavenged on trout eggs in the field.
5. There was some evidence that Rhyacophila dorsalis (Curtis) and Diura bicaudata (L.) preyed on trout eggs, but in view of their scarcity, it is suggested that they were a minor mortality factor.     

Viability and acrosome staining of stallion spermatozoa by Chicago sky blue and Giemsa.

A simple trypan blue-neutral red-Giemsa staining procedure for simultaneous evaluation of acrosome, sperm head, and tail membrane integrity and morphology has been used to evaluate equine spermatozoa. Some special characteristics and problems have arisen in evaluating stallion semen. One problem was the differentiation of intact vs. damaged sperm tails primarily in frozen and thawed samples. After freezing and thawing, a high percentage of spermatozoa with an unstained head and stained tail were observed. These cells are considered immotile. Therefore, unambiguous differentiation of intact vs. damaged sperm tail membrane is very important for evaluating semen quality. The aim of our study was to develop a method especially for stallion sperm to distinguish more accurately the different cell types. We compared Chicago sky blue 6B (CSB) to trypan blue (TB) for viability staining. CSB/Giemsa staining showed good repeatability and agreement with TB/Giemsa measurements. For densitometry analysis, individual digital images were taken from smears stained by CSB/Giemsa and by TB/Giemsa. A red-green-blue (RGB) histogram for each area of spermatozoa was drawn. Differences of means of RGB values of live vs. dead tails and separate live vs. dead heads from each photo were used to compare the two staining procedures. CSB produced similar live/dead sperm head differentiation and better tail differentiation. TB can be replaced by CSB and this results in more reliable evaluation. After staining with 0.16% CSB and 4 min fixation, 2-4 h Giemsa staining at 25-40 degrees C is recommended for stallion semen.