A Bacterial Swimmer with Two Alternating Speeds of Propagation

We recorded large data sets of swimming trajectories of the soil bacterium Pseudomonas putida. Like other prokaryotic swimmers, P. putida exhibits a motion pattern dominated by persistent runs that are interrupted by turning events. An in-depth analysis of their swimming trajectories revealed that the majority of the turning events is characterized by an angle of ϕ₁ = 180° (reversals). To a lesser extent, turning angles of ϕ₂ = 0° are also found. Remarkably, we observed that, upon a reversal, the swimming speed changes by a factor of two on average—a prominent feature of the motion pattern that, to our knowledge, has not been reported before. A theoretical model, based on the experimental values for the average run time and the rotational diffusion, recovers the mean-square displacement of P. putida if the two distinct swimming speeds are taken into account. Compared to a swimmer that moves with a constant intermediate speed, the mean-square displacement is strongly enhanced. We furthermore observed a negative dip in the directional autocorrelation at intermediate times, a feature that is only recovered in an extended model, where the nonexponential shape of the run-time distribution is taken into account.     

Analyzing fish movement as a persistent turning walker

The trajectories of Kuhlia mugil fish swimming freely in a tank are analyzed in order to develop a model of spontaneous fish movement. The data show that K. mugil displacement is best described by turning speed and its auto-correlation. The continuous-time process governing this new kind of displacement is modelled by a stochastic differential equation of Ornstein–Uhlenbeck family: the persistent turning walker. The associated diffusive dynamics are compared to the standard persistent random walker model and we show that the resulting diffusion coefficient scales non-linearly with linear swimming speed. In order to illustrate how interactions with other fish or the environment can be added to this spontaneous movement model we quantify the effect of tank walls on the turning speed and adequately reproduce the characteristics of the observed fish trajectories.     

Changes in the swimming behavior of Pseudodiaptomus annandalei (Copepoda,Calanoida) adults exposed to the diatom toxin 2-trans, 4-trans decadienal

Diatoms are broadly present in marine habitats and often dominate seasonal phytoplankton blooms in polar and temperate latitudes. Certain species produce polyunsaturated aldehydes upon mechanical wounding caused by mesozooplankton grazing. Ample evidence is available on toxin-induced reproductive failure in copepods, yet their behavioral effects remain unclear. Here we present results of laboratory experiments in which we investigated the immediate effects of the diatom-derived aldehyde 2-trans, 4-trans decadienal on the three-dimensional swimming behavior of the calanoid copepod Pseudodiaptomus annandalei. Short-term direct exposure to the toxin at 3 μM, 6 μM and 12 μM induced hyperactivity in the three adult states, as evidenced by a marked and dose-dependent increase in the number of trajectories. It also caused a higher proportion of vertical movements. In males and ovigerous females exposed to decadienal at 3 μM and 6 μM, hyperactivity came with an equally specific dose-dependent decrease in swimming speed. Males and ovigerous females swam faster at 12 μM than at 6 μM, suggesting a complex mode of action of the toxin. In non-ovigerous females, decadienal had little effects on swimming speed, supporting the assumption that female copepods are less affected by certain environmental stressors. Multifractal analysis revealed differences in the statistical properties of the swimming behavior between experimental conditions. The moment structure function of the displacement appeared to be moderately multifractal in the three adult states swimming in control water. Ethanol as carrier solvent at 200 ppm caused an increase in swimming speed and a switch toward a more ballistic motion in males and ovigerous females. On the opposite, exposure to the toxin reduced or cancelled the effects of ethanol and resulted in a more Brownian motion for high moment values. Decadienal had little effects on the behavior of non-ovigerous females except at the highest concentration. Our results demonstrate that decadienal, a model diatom aldehyde, impairs the behavior of adult copepods. They provide further information on the interaction between diatoms and their main predator.     

Ontogenetic changes in the critical swimming speed of Gadus morhua (Atlantic cod) and Myoxocephalus scorpius (shorthorn sculpin) larvae and the role of temperature

Most studies on behavioural contributions to dispersal and recruitment during early life history stages of fishes have focused on coral reef species. For cold ocean environments, high variation in seasonal temperature and development times suggest that parallel studies on active behaviour are needed for cold-water species. Thus, we examined the critical swimming speed (U_crit) of marine fish larvae from 2 contrasting species: Gadus morhua (Atlantic cod) and Myoxocephalus scorpius (shorthorn sculpin), a pelagic and bottom spawner respectively. Within-species comparisons showed that sculpin reared at 6 °C had lower initial U_crit values, but a faster U_crit increase through development compared with 3 °C conspecifics, ultimately resulting in faster critical swimming speeds at metamorphosis (10.5 vs. 9.1 cm·s^− 1). In contrast, although cod larvae reared at 10 °C were faster swimmers at first feeding than 6 °C fish, temperature differences were absent after the first week. These results show that temperature influences the trajectory of larval critical swimming speed development, but that the relationship is species-specific. Although 6 °C sculpin and cod of similar length had equivalent U_crit values, the smaller size of cod at hatch (5.3 vs. 10.8 mm for sculpin) resulted in much lower age-specific U_crit values for cod. These data have significant implications for how swimming activity of the two species might affect dispersal, particularly in the first few weeks post-hatch. Overall, our data suggest that temperature during larval development influences the swimming capacity of cold-water marine fishes, and has important ramifications for biophysical models of dispersal.     

The effects of feeding on the swimming performance and metabolic response of juvenile southern catfish,Silurus meridionalis,acclimated at different temperatures

To test whether the effects of feeding on swimming performance vary with acclimation temperature in juvenile southern catfish (Silurus meridionalis), we investigated the specific dynamic action (SDA) and swimming performance of fasting and feeding fish at acclimation temperatures of 15, 21, 27, and 33 °C. Feeding had no effect on the critical swimming speeding (U_crit) of fish acclimated at 15 °C (p = 0.66), whereas it elicited a 12.04, 18.70, and 20.98% decrease in U_crit for fish acclimated at 21, 27 and 33 °C, respectively (p < 0.05). Both the maximal postprandial oxygen consumption rate (VO_2peak) and the active metabolic rate (VO_2active, maximal aerobic sustainable metabolic rate of fasting fish) increased significantly with temperature (p < 0.05). The postprandial maximum oxygen consumption rates during swimming (VO_2max) were higher than the VO_2active of fasting fish at all temperature groups (p < 0.05). The VO_2max increased with increasing temperature, but the relative residual metabolic scope (VO_2max? VO_2peak) during swimming decreased with increasing in temperature. The present study showed that the impairment of postprandial swimming performance increased with increasing temperature due to the unparalleled changes in the catfish's central cardio-respiratory, peripheral digestive and locomotory capacities. The different metabolic strategies of juvenile southern catfish at different temperatures may relate to changes in oxygen demand, imbalances in ion fluxes and dissolved oxygen levels with changes in temperature.     

The use of spontaneous behavior,swimming performances and metabolic rate to evaluate toxicity of PFOS on topmouth gudgeon Pseudorasbora parva

Perfluorooctane sulfonate (PFOS) is a ubiquitous environmental contaminant that has been found to pose various risks to fish health and the safety of aquatic ecosystem. Swimming performance is an integrated index of fitness in fish. However, little research has sought on the effects of PFOS on swimming performances of fish. Experiments were carried out to clarify the impacts of acute exposure to PFOS on behavior, swimming ability and metabolic rate in topmouth gudgeon (Pseudorasbora parva), to understand the underlying ecotoxicological effects of waterborne PFOS exposure on fish physiology and behavior. Fish were exposed to PFOS (0, 0.5, 2, 8 or 32 mg/L) for 96 h. Afterwards, the routine metabolic rate (RMR), spontaneous swimming behavior (SSB), fast-start swimming performance (FSP) and critical swimming speed (U_crit) of the topmouth gudgeon were examined. The results show reduced behavioral performance and increased physiological stress with increasing PFOS concentration. Both RMR, SSB and U_crit were significantly affected by PFOS exposure (p < 0.05). The lowest observed effect concentration (LOEC) is 2 mg/L for SSB. PFOS treatment resulted in increased RMR (p = 0.001) and decreased U_crit (p = 0.005), whereas FSP was not influenced by PFOS (p > 0.05). The results indicate that the anaerobic swimming capacity was conserved, but the metabolic level, SSB and aerobic swimming performance in topmouth gudgeon were susceptible to PFOS contamination, and hence might be useful as considerable potential biomarkers of pollution.     

Switching of Swimming Modes in Magnetospirillium gryphiswaldense

The microaerophilic magnetotactic bacterium Magnetospirillum gryphiswaldense swims along magnetic field lines using a single flagellum at each cell pole. It is believed that this magnetotactic behavior enables cells to seek optimal oxygen concentration with maximal efficiency. We analyze the trajectories of swimming M. gryphiswaldense cells in external magnetic fields larger than the earth’s field, and show that each cell can switch very rapidly (in <0.2 s) between a fast and a slow swimming mode. Close to a glass surface, a variety of trajectories were observed, from straight swimming that systematically deviates from field lines to various helices. A model in which fast (slow) swimming is solely due to the rotation of the trailing (leading) flagellum can account for these observations. We determined the magnetic moment of this bacterium using a to our knowledge new method, and obtained a value of (2.0 ± 0.6) × 10^?16 A · m². This value is found to be consistent with parameters emerging from quantitative fitting of trajectories to our model.     

Hypoxic avoidance behaviour in cod (Gadus morhua L.): The effect of temperature and haemoglobin genotype

Hypoxia can influence fish growth, survival and on larger scales, population structure. These effects may be influenced by water temperature, and may vary intra-specifically with genotype. In Atlantic cod (Gadus morhua L.), the two haemoglobin homozygotes (Hb-I?11 and Hb-I?22) vary in oxygen affinity at different temperatures, which is thought to correspond to variation in hypoxia tolerance. We therefore tested if hypoxic avoidance behaviour in cod 1) depends on ambient temperature and 2) is modified by haemoglobin genotype. In a laminar flow choice box, we subjected juvenile cod to an initial phase of non-escapable hypoxia, and a subsequent recovery phase, where one habitat was kept at 20% O₂ saturation while the other was raised in steps to full saturation. The experiment was performed at 5 and 15 °C with Hb-I?11 and Hb-I?22 cod. Cod responded to inescapable hypoxia by reducing their overall swimming speed and then, at the initial levels of the recovery phase, avoiding the most hypoxic habitat, irrespective of temperature or genotype. Fish recovered quickly as O₂ levels increased, as evidenced by increased swimming speed and time spent in the most hypoxic habitat. The avoidance response depended strongly on temperature: the relative reduction in speed and avoidance of the most hypoxic habitat was more pronounced at 15 than at 5 °C. During the recovery phase, stressed fish initially maintained a higher swimming speed in the most hypoxic habitat. However, as O₂ increased, swimming speed in both habitats converged. This point of convergence occurred at a lower O₂ saturation at 5 °C. Fish ventilation rate in inescapable hypoxia was also higher at 15 °C. Haemoglobin genotype did not influence either ventilation rates or the nature of the hypoxic avoidance response at either temperature, but Hb-I?11 cod swam faster than Hb-I?22 cod in normoxia at 15 °C. Our results indicate that increased temperature limits the ability of cod of both haemoglobin genotypes to exploit hypoxic habitats. This may have negative future consequences for coastal cod stocks in light of increasing global temperatures and eutrophication in coastal waters.     

Catechol 1,2-dioxygenase from the new aromatic compounds - Degrading Pseudomonas putida strain N6

This study aimed to characterization of catechol 1,2-dioxygenase from a Gram-negative bacterium, being able to utilize a wide spectrum of aromatic substrates as a sole carbon and energy source. Strain designated as N6, was isolated from the activated sludge samples of a sewage treatment plant at Bentwood Furniture Factory Jasienica, Poland. Morphology, physio-biochemical characteristics and phylogenetic analysis based on 16S rDNA sequence indicate that strain belongs to Pseudomonas putida. When cells of strain N6 grown on protocatechuate or 4-hydroxybenzoic acid mainly protocatechuate 3,4-dioxygenase was induced. The activity of catechol 1,2-dioxygenase was rather small. The cells grown on benzoic acid, catechol or phenol showed high activity of only catechol 1,2-dioxygenase. This enzyme was optimally active at 35 °C and pH 7.4. Kinetic studies showed that the value of K_m and V_max was 85.19 ??M and 14.54 ??M min⁻¹ respectively. Nucleotide sequence of gene encoding catechol 1,2-dioxygenase in strain N6 has 100% identity with catA genes from two P. putida strains. The deduced 301-residue sequence of enzyme corresponds to a protein of molecular mass 33.1 kDa. The deduced molecular structure of the catechol 1,2-dioxygenase from P. putida N6 was very similar and characteristic for the other intradiol dioxygenases.     

Mechanism of cyanide and thiocyanate decomposition by an association of Pseudomonas putida and Pseudomonas stutzeri strains

The intermediate and terminal products of cyanide and thiocyanate decomposition by individual strains of the genus Pseudomonas, P. putida strain 21 and P. stutzeri strain 18, and by their association were analyzed. The activity of the enzymes of nitrogen and sulfur metabolism in these strains was compared with that of the collection strains P. putida VKM B-2187^T and P. stutzeri VKM B-975^T. Upon the introduction of CN⁻ and SCN⁻ into cell suspensions of strains 18 and 21 in phosphate buffer (pH 8.8), the production of NH ₄ ⁺ was observed. Due to the high rate of their utilization, NH₃, NH ₄ ⁺ , and CNO⁻ were absent from the culture liquids of P. putida strain 21 and P. stutzeri strain 18 grown with CN⁻ or SCN⁻. Both Pseudomonas strains decomposed SCN⁻ via cyanate production. The cyanase activity was 0.75 μmol/(min mg protein) for P. putida strain 21 and 1.26 μmol/(min mg protein) for P. stutzeri strain 18. The cyanase activity was present in the cells grown with SCN⁻ but absent in cells grown with NH ₄ ⁺ . Strain 21 of P. putida was a more active CN⁻ decomposer than strain 18 of P. stutzeri. Ammonium and CO₂ were the terminal nitrogen and carbon products of CN⁻ and SCN⁻ decomposition. The terminal sulfur products of SCN⁻ decomposition by P. stutzeri strain 18 and P. putida strain 21 were thiosulfate and tetrathionate, respectively. The strains utilized the toxic compounds in the anabolism only, as sources of nitrogen (CN⁻ and SCN⁻) and sulfur (SCN⁻). The pathway of thiocyanate decomposition by the association of bacteria of the genus Pseudomonas is proposed based on the results obtained. Original Russian Text ? N.V. Grigor’eva, T.F. Kondrat’eva, E.N. Krasil’nikova, G.I. Karavaiko, 2006, published in Mikrobiologiya, 2006, Vol. 75, No. 3, pp. 320–328.     

Pectoral fin beat frequency predicts oxygen consumption during spontaneous activity in a labriform swimming fish (Embiotoca lateralis)

The objective of this study was to identify kinematic variables correlated with oxygen consumption during spontaneous labriform swimming. Kinematic variables (swimming speed, change of speed, turning angle, turning rate, turning radius and pectoral fin beat frequency) and oxygen consumption (MO₂) of spontaneous swimming in Embiotoca lateralis were measured in a circular arena using video tracking and respirometry, respectively. The main variable influencing MO₂ was pectoral fin beat frequency (r ² = 0.71). No significant relationship was found between swimming speed and pectoral fin beat frequency. Complementary to other methods within biotelemetry such as EMG it is suggested that such correlations of pectoral fin beat frequency may be used to measure the energy requirements of labriform swimming fish such as E. lateralis in the field, but need to be taken with great caution since movement and oxygen consumption patterns are likely to be quite different in field situation compared to a small lab tank. In addition, our methods could be useful to measure metabolic costs of growth and development, or bioassays for possible toxicological effects on fish.     

Purification,kinetic and thermodynamic characterization of soluble acid invertase from sugarcane (Saccharum officinarum L.)

We report for the first time kinetic and thermodynamic properties of soluble acid invertase (SAI) of sugarcane (Saccharum officinarum L.) salt sensitive local cultivar CP 77-400 (CP-77). The SAI was purified to apparent homogeneity on FPLC system. The crude enzyme was about 13 fold purified and recovery of SAI was 35%. The invertase was monomeric in nature and its native molecular mass on gel filtration and subunit mass on SDS-PAGE was 28 kDa. SAI was highly acidic having an optimum pH lower than 2. The acidic limb was missing. Proton transfer (donation and receiving) during catalysis was controlled by the basic limb having a pKa of 2.4. Carboxyl groups were involved in proton transfer during catalysis. The kinetic constants for sucrose hydrolysis by SAI were determined to be: k_m = 55 mg ml^?1, k_cat = 21 s^?1, k_cat/k_m = 0.38, while the thermodynamic parameters were: ΔH* = 52.6 kJ mol^?1, ΔG* = 71.2 kJ mol^?1, ΔS* = ?57 J mol^?1 K^?1, ΔG*_E–S = 10.8 kJ mol^?1 and ΔG*_E–T = 2.6 kJ mol^?1. The kinetics and thermodynamics of irreversible thermal denaturation at various temperatures 53–63 °C were also determined. The half -life of SAI at 53 and 63 °C was 112 and 10 min, respectively. At 55 °C, surprisingly the half -life increased to twice that at 53 °C. ΔG*, ΔH* and ΔS* of irreversible thermal stability of SAI at 55 °C were 107.7 kJ mol^?1, 276.04 kJ mol^?1 and 513 J mol^?1K^?1, respectively.     

Production of two monomer structures containing medium-chain-length polyhydroxyalkanoates by β-oxidation-impaired mutant of Pseudomonas putida KT2442

Pseudomonas putida KT2442 produces medium-chain-length (MCL) polyhydroxyalkanoates (PHA) from fatty acids. When gene encoding 3-hydroxyacyl-CoA dehydrogenase which catalyzes long-chain-3-hydroxyacyl-CoA to 3-ketoacyl-CoA, was partially or completely deleted in P. putida KTOY08, the PHA accumulated was shown to contain only two different monomer structures dominated by a monomer of the same chain length as that of the fatty acids fed and another monomer two carbon atoms shorter. Among the PHA copolymers, P(44% 3HD-co-3HDD) containing 44% 3HD and 56% 3HDD was demonstrated to have a melting temperature T_m, an apparent heat of fusion △H_m and a Young’s modulus E of 75 °C, 51 J g^?1 and 2.0 MPa, respectively, the highest among all the MCL PHA studied.     

Effects of acute temperature changes on the swimming abilities and oxygen consumption of Ptychobarbus kaznakovi from the Lancang River

Water temperature is known to be a particularly important environmental factor that affects fish swimming performance, but it is unknow how acute temperature changes affect the fish performance of Ptychobarbus kaznakovi. P. kaznakovi in the Lancang River have declined quickly in recent years, and this species was used to examine the effects of acute temperature changes on swimming abilities and oxygen consumption in a Brett‐type swimming tunnel respirometer. The standard metabolic rate (SMR) and routine metabolic rate (RMR) showed 216% and 134% increases, respectively, at 22°C (an acute increase from 17 to 22°C) compared to those at 12°C (an acute decrease from 17 to 12°C). Moreover, the RMR was approximately 1.7, 1.6 and 1.3 times the value of the SMR at 12°C, 17°C and 22°C, respectively. The critical swimming speed (U_crit) of P. kaznakovi at 22°C was 5.45 ± 0.45BL/S, which was 45% higher than that at 12°C (3.77 ± 0.92BL/S). The oxygen consumption rates (MO₂) reached their maximum values at swimming speeds near the U_crit for all the temperature treatments. The maximum metabolic rate (MMR) values at 12°C, 17°C and 22°C were 274.53 ± 142.60 (mgO₂ kg^?1 hr^?1), 412.85 ± 216.34 (mgO₂ kg^?1 hr^?1) and 1,095.73 ± 52.50 (mgO₂ kg^?1 hr^?1), respectively. Moreover, there was a narrow aerobic scope at 12°C compared to that at 17°C and 22°C. The effect of acute temperature changes on the swimming abilities and oxygen consumption of P. kaznakovi indicated that water temperature changes caused by dam construction could directly affect energy consumption during the upstream migration of fish.     

The Zinc Center Influences the Redox and Thermodynamic Properties of Escherichia coli Thioredoxin 2

Thioredoxins are small, ubiquitous redox enzymes that reduce protein disulfide bonds by using a pair of cysteine residues present in a strictly conserved WCGPC catalytic motif. The Escherichia coli cytoplasm contains two thioredoxins, Trx1 and Trx2. Trx2 is special because it is induced under oxidative stress conditions and it has an additional N-terminal zinc-binding domain. We have determined the redox potential of Trx2, the pK_a of the active site nucleophilic cysteine, as well as the stability of the oxidized and reduced form of the protein. Trx2 is more oxidizing than Trx1 (-221 mV versus -284 mV, respectively), which is in good agreement with the decreased value of the pK_a of the nucleophilic cysteine (5.1 versus 7.1, respectively). The difference in stability between the oxidized and reduced forms of an oxidoreductase is the driving force to reduce substrate proteins. This difference is smaller for Trx2 (ΔΔG°_H2O = 9 kJ/mol and ΔT_m = 7. 4 °C) than for Trx1 (ΔΔG°_H2O = 15 kJ/mol and ΔT_m = 13 °C). Altogether, our data indicate that Trx2 is a significantly less reducing enzyme than Trx1, which suggests that Trx2 has a distinctive function. We disrupted the zinc center by mutating the four Zn²⁺-binding cysteines to serine. This mutant has a more reducing redox potential (-254 mV) and the pK_a of its nucleophilic cysteine shifts from 5.1 to 7.1. The removal of Zn²⁺ also decreases the overall stability of the reduced and oxidized forms by 3.2 kJ/mol and 5.8 kJ/mol, respectively. In conclusion, our data show that the Zn²⁺-center of Trx2 fine-tunes the properties of this unique thioredoxin.     

Reduced scapular muscle control and impaired shoulder joint position sense in subjects with chronic shoulder stiffness

The purpose of the present study was to determine (1) if joint position sense (JPS) in subjects with shoulder stiffness (SS) differs from that in controls; (2) if, when JPS is reduced in SS, it is related to scapular muscular activities in the mid/end ranges of motion; and (3) if a person’s function is associated with his or her level of JPS. Eighteen subjects with unilateral SS and 18 controls were included. Each subject performed abduction by self-selecting an end/mid range position. The electromagnetic motion-capturing system collected kinematic data while surface electromyography collected muscle activities (upper trapezius, lower trapezius, and serratus anterior muscles). Subjects were asked to move the upper limb to the target position (end/mid range) accurately without visual guidance. Reduced JPS was observed in subjects with SS (2.7 degrees in mid range, p < 0.05). The JPS was enhanced by an increased scapula muscular activation level in the end range of motion (R = ?0.61 for SS and ?0.41 for controls) and by coordination among muscles’ activation in the mid-range of motion (R = ?0.87 for SS and R = ?0.53 for controls). Impaired JPS was also related to self-reported functional status (R = ?0.56) in subjects with SS. Shoulder JPS in subjects with chronic SS is impaired in comparison with controls. In the mid-range motion, the coordination of scapula muscular activation is related to shoulder JPS. Impaired JPS is also function-related in subjects with SS. These findings suggest that the coordination among scapula muscles’ activation were important to consider in the rehabilitation of patients with chronic SS.