Toxicity of Cadmium to Amoeba Proteus: A Biochemical Approach

SYNOPSIS The cadmium ion (Cd²⁺) was accumulated by Amoeba proteus in all cellular fractions, the highest level being associated with the cytosol fraction. On gel separation of the cytosol fraction, Cd-binding protein appeared in 2 peaks: one >45,000 MW (peak I) and the other 12,000 MW (peak II). Added cysteine increased the total Cd²⁺ taken up by the cells and resulted in disproportionate increase of Cd incorporated into the Cd-binding protein of peak II. the Cd-binding protein of peak II is analogous to the low-MW, Cdbinding proteins in Anacystis nidulans, Mytilus edulis , and to the metalloprotein of some vertebrates.     

The Contractile Vacuole in Amoeba proteus: Temperature Effects

The influence of temperature on the various aspects of the contractile vacuole cycle of Amoeba proteus has been established. In the upper temperature range (20, 25 and 30 C) an increase in temperature results in shorter vacuolar cycles with greater systolic (final) volumes. The systole is rapid and always complete. At 35 C the vacuole shows the effect of heat stress, cycles are irregular in volume and duration with only partial systoles. In the lower temperature range (15, 10 and 5 C), a new phenomenon has been observed, the plateau. Instead of undergoing systole, after reaching a certain critical volume the vacuole abruptly ceases to grow in size and remains in a state of pause for a well defined period of time, ending at a comparatively slow but complete systole. The duration of this plateau as well as its inception and termination seem quite precisely controlled. Its effect, a decrease in the fluid output by the vacuole, is such as to adjust vacuolar output to near constant Q₁₀ kinetics over our temperature range. This is correlated with a single straight line fit in an Arrhenius plot. Available data do not permit a complete explanation of the nature of the plateau. It could represent a steady state between 2 opposing phenomena: active fluid influx into the vacuole and osmotic losses from the vacuole into the relatively hypertonic cytoplasm.     

The cytoskeleton of spreadingDictyostelium amoebae

Summary The cytoarchitectural elements ofDictyostelium discoideum amoeba have been visualized by light and electron microscopy in cells prepared with mixtures of glutaraldehyde and Triton-X-100. After negative staining, the peripheral regions of spreading amoebae show a complex meshwork of actin filaments, the majority of which were less than 0.25 microns in length. Multiple branch points, end to side abutments and cross-overs were characteristic features of the actin meshworks. Filopodia extending from the cell periphery consisted of bundles of actin filaments that penetrated into and merged with the actin meshworks in the spreading lamellae. Microtubules emanating from the nucleus associated body penetrated to differing extents into the actin meshworks, sometimes extending close to the cell periphery.Dictyostelium cytoskeletons preparted as described here should prove useful for further studies on the locomotory mechanism.     

Scanning Electron Microscope Observations of Amoeba proteus During Phagocytosis*

SYNOPSIS. Phagocytosing Amoeba proteus at different stages of forming foodcups have been observed by scanning electron microscopy. A nonphagocytosing ameba is characterized by dorsal and lateral ridges running longitudinally over the posterior half of the cell and its attachment to the substrate over small areas. When stimulated by prey organisms, the ameba loses polarity and ridges, and adheres to the substrate more firmly over a wider area of contact. Then it forms broad pseudopods to surround its prey and this results in the formation of foodcups. The surface of all amebae is covered with small projections, and membranous blebs are often seen on the surface of phagocytosing organisms.     

Resuscitation of amebae deprived of essential symbiotes: Micrurgical studies*

SYNOPSIS. The effect of depletion and restoration of obligatory bacterial endosymbiotes on Amoeba proteus strain xD was studied. Removal of the symbiotes by culturing the amebae at 26.5 C resulted in loss of viability of the host cells, indicating that this strain is dependent on its endosymbiotes for survival. Amebae depleted of bacteria could initially be resuscitated by injection of isolated symbiotes, but prolonged deprivation led to irreversible changes. Nuclei of aposymbiotic amebae were viable when transplanted into the cytoplasm of normal cells, but the symbiote-depleted cytoplasm of heat-treated amebae could not be resuscitated by renucleation. No immediate ultrastructural changes were detected in aposymbiotic amebae except for clumping of nucleoli. Thus it appears that the symbiote performs an essential function as a cytoplasmic constituent.     

The multiplication dynamics of pathogenic bacteria in relation to the trophic and temperature cultivation conditions

The comparative study of the dynamics of multiplication of Listeria monocytogenes and Yersinia pseudotuberculosis in organic and synthetic media and in distilled water at temperatures of 37 degrees C and 6 degrees C was carried out. This study revealed that in organic media the multiplication of bacteria was good at 37 degrees C and 6 degrees C. In mineral media and distilled water their multiplication was observed only at 6 degrees C. Moreover, conditions necessary for the multiplication of pathogenic bacteria in distilled water were shown; these conditions depended on the inoculated dose, the number of autolyzed microbial cells and the state of the culture. Proofs of the multiplication of the bacteria under the conditions of minimum nutrition and low temperature were obtained with the use of labeled 3H-thymidine.     

Effect of Chloramphenicol on Bacterial Endosymbiotes in a Strain of Amoeba proteus*

SYNOPSIS. The effect of chloramphenicol (CAP) on the bacterial endosymbiotes of a strain of Amoeba proteus was studied by growing the symbiotic amebae in media containing 0.5–1.6 mg/ml CAP for up to 4 weeks. Treatments with CAP caused such ultrastructural changes as expansion of the nuclear zone and deformation of symbiotes. The CAP treatment also damaged the mitochondria, e.g. disappearance of central and protrusion of peripheral cristae. Number of bacteria per ameba decreased to < 10% of control in CAP-containing media, but no viable amebae became completely free of symbiotes. The resuts supported previous studies that amebae were dependent on endosymbiotes.     

变温对两种昆虫发育速率的影响

报道了松毛虫赤眼蜂和瓜螟在多组恒温、交替变温下的发育历期。对结果的分析表明,温度交替对这两种昆虫在任一温度下的瞬时发育率无明显影响,恒温下和变温下完成发育所需的热量无显著差异或基本相似。     

The Fine Structure of Four “Species” of Amoeba*

The fine structure of Amoeba discoides, Amoeba dubia, and Amoeba amazonas was studied and compared with that of Amoeba proteus. The different kinds of amebas showed general similarities but differed in the ultrastructural details of their organelles. With respect to fine structure, A. discoides was indistinguishable from A. proteus, while both A. dubia and A. amazonas had distinctive features. The nuclei of all had a prominent honeycomb-like fibrous lamina, but A. dubia differed from the others in the distribution of nucleoli within the nucleus. The mitochondria of A. amazonas were unusual in having a variable pattern of cristae, some being plate-like and others tubular. Golgi bodies in A. amazonas had a greater proportion of vesicles and a smaller number of cisternae than those of the others, while Golgi bodies in A. dubia had highly flattened cisternae without a lining of filamentous material such as is found in the other types. The plasma membrane of A. dubia also lacked the prominent filamentous cell coat common to A. proteus and other amebas. The relation between the Golgi apparatus and the cell coat and the significance of the degree of development of the cell coat for pinocytosis and other phenomena is considered. The experimental use of these cells, including the formation of hybrids by nuclear transplantation is discussed.     

Enhanced rate of multiplication and rooting through the use of coir in aseptic liquid culture media

In the quest for cheap and suitable eco-friendly matrix with liquid culture to facilitate rooting three matrices, viz. paddy straw, jute and coir were chosen. Of the three matrices, coir was selected due to its higher water retention capacity compared to the other two matrices. Coir was used with aseptic liquid media for rooting of ten plant species, viz., Nicotiana tabacum, Andrographis paniculata, Chenopodium album (2n=54), C. album (Broad leaf diploid), C. album (Narrow leaf diploid), C. murale, Beta vulgaris (Sugar beet and Table beet), B. palonga (Indian spinach), Tectona grandis and Musa spp. Higher number of roots with profuse root hairs were recorded in all the plant materials in liquid medium with coir compared to conventional agar-gelled media. Apart from rooting, the rate of multiplication, maintenance and concomitant hardening were also found to be more effective in comparison to agar-gelled media. The economical aspect of use of coir instead of agar is also discussed.     

The influence of thermal conditions on rectal temperature,respiration rate and pulse rate of lactating Holstein-Friesian cows in the humid tropics

The effect of minimum, maximum and mean ambient air temperatures and the temperature-humidity index (THI) of the same and the previous day on morning (a.m.) and afternoon (p.m.) rectal temperatures (RT), respiration rates (RR) and pulse rates (PR) were studied in 17 Holstein-Friesian cows over the first 125 days in the 3rd and 4th lactations. Physiological responses showed a diurnal pattern, being lower in the mornings than the afternoons: 38.6 vs 39.0° C for RT, 52.2 vs 60.7 breaths/min for RR and 58.1 vs 64.1 beats/min for PR. Correlations between RT and RR (r = – 0.043 to –0.046) and RT and PR (r = –0.178 to –0.261) were low (P> 0.05). Correlations between RR and PR (r = 0.353 to 0.365) were moderate (P<0.05). Weather variables, especially ambient temperature of the previous day, were more important and influenced physiological responses to a greater extent than other thermal factors the same day. Generally, physiological responses were influenced to a greater extent by ambient temperature than THI. Weather variables explained variations in RT (5.1–59.6%), in RR (13.0–17.8%) and in PR (22.1–25.4%). Relationships between weather variables the previous day and physiological responses were contradictory, with minimum and maximum values showing a negative relationship in contrast with a positive relationship for mean values.     

The necessity for low temperature treatment on the eggs of Emma Cricket,Teleogryllus emma and the method of increasing the hatching rate after low temperature treatment

The necessity for low temperature treatment of Emma Cricket Eggs, Teleogryllus emma, to dormancy breaking and the method of increasing hatching rate after low temperature treatment were investigated. Emma Cricket Eggs were treated by constant temperature, room temperature, low temperature, step‐down low temperature. As a result, constant temperature treatment was the lowest on hatching rate. The hatching rates of the rest of other temperature treatments were not different statistically (one‐way ANOVA). In other word, low temperature treatment was not necessary to dormancy breaking. But low temperature treatment was necessary to control individual number of Emma Cricket, Teleogryllus emma, on the mass indoor‐rearing system. And the step‐up temperature treatment was necessary to increase hatching rate after low temperature treatment.     

Ultra-structure and localisation of formazan formed by human neutrophils and amoebae phagocytosing virulent and avirulent Legionella pneumophila

Abstract Legionella pneumophila (LP) strains of differing virulence were incubated with a solution of nitroblue-tetrazolium (NBT) at a concentration of 1 mg · ml⁻ in the presence of Acanthamoeba polyphaga or human polymorphonuclear neutrophils (PMN). Reduction of NBT to formazan occurred at a faster rate in the presence of virulent strains. Reduction appeared to be temperature dependent; at 37°C the reaction rate was higher than at 20°C. On microscopic examination, deposits of formazan around Legionella cells were observed inside amoebae similar to those deposited in human neutrophils. Electron microscopy revealed electron-dense particles surrounding virulent legionellae, which appeared to be associated with formazan foundation. Formazan formation inside amoebae may suggest the presence of a respiratory burst against LP, which is more intense with virulent strains.     

Characteristics of trajectory in the migration of Amoeba proteus

Summary. We investigated the behavior of migration of Amoeba proteus in an isotropic environment. We found that the trajectory in the migration of A. proteus is smooth in the observation time of 500-1000 s, but its migration every second (the cell velocity) on the trajectory randomly changes. Stochastic analysis of the cell velocity and the turn angle of the trajectory has shown that the histograms of the both variables well fit to Gaussian curves. Supposing a simple model equation for the cell motion, we have estimated the motive force of the migrating cell, which is of the order of piconewton. Furthermore, we have found that the cell velocity and the turn angle have a negative cross-correlation coefficient, which suggests that the amoeba explores better environment by changing frequently its migrating direction at a low speed and it moves rectilinearly to the best environment at a high speed. On the other hand, the model equation has simulated the negative correlation between the cell velocity and the turn angle. This indicates that the apparently rational behavior comes from intrinsic characteristics in the dynamical system where the motive force is not torquelike.     

阿里山潜蝇茧蜂的羽化率及死亡率与储存温度的拟合关系

本文通过室内模拟低温,研究了阿里山潜蝇茧蜂雌成蜂、9日龄蛹的致死中温度和线性拟合。结果表明,在不同温度下雌蜂的砜分别为4℃下6．6149天,7℃下8．1235天,100℃下9．6161天,13℃下5．0257天,且在所有温度下都有死亡率陡升过程。9日龄蛹在不同温度下的LT50分别为4℃下8．4172天,7℃下8．1235天,10℃下13．9949天,13℃下18．4107天;同一温度下的蛹羽化率与保存天数的关系适宜用二次方程拟合。研究认为蛹的耐寒性强于雌成蜂,可以作为低温保存的虫态。     

The metabolic rate of roach in relation to body size and temperature

Standard and routine metabolic rates of roach Rutilus rutilus for a wide size and temperature range (3–200 g, 5–23° C) were analysed by automated, computerized intermittent flow respirometry. The mass exponent b ranged from 0·68 to 0·82 for standard metabolism, and from 0·65 to 0·92 for routine metabolism depending on the experimental temperature. For routine metabolism b was lowest at 10° C. At both decreasing and increasing temperatures, b increased significantly. Roach were exponentially temperature-dependent for both metabolic levels. For roach <20 g, however, an asymptotic relationship was observed between temperature and routine metabolic rate. The 'flattening of the curve' in the latter case may be explained by reduced spontaneous activities at the lower threshold of the preferred temperature range.     

温度和个体大小对单齿螺耗氧率和排氨率的影响

采用静水法测定了不同温度、不同个体大小的单齿螺耗氧率和排氨率。结果表明:在16-33℃的实验温度范围内单齿螺的耗氧率(R_O)和排氨率(R_N)与软体部干重(W)都呈负相关,它们之间关系可以分别用幂函数R_O=aW^-b和R_N=a₁W^-b1表示。16～29℃温度范围内单齿螺的耗氧率和排氨率均随温度的升高而增加,29℃时耗氧率和排氨率达到最大值,当温度继续升高超过29℃后,耗氧率和排氨率则随温度的升高而下降,耗氧率、排氨率与温度之间呈显著的指数函数关系R_O=ce^dT和R_N=c₁e^d1T;不同个体大小单齿螺的O:N比在16～20℃时较大,Q₁₀取值范围0.56-3.74,平均值为1.64。方差分析表明,温度、软体部干重对单齿螺的耗氧率和排氨率均有极显著的影响(P<0.01)。     

保存时间及温度对大肠杆菌感受态转化效率的影响

通过不同保存时间和温度对大肠杆菌感受态转化效率影响的研究,更明确阐明了不同保存温度下感受态细胞随保存时间的延长其转化率的变化趋势,为以后的研究提供了量化的指标。