Effects of temperature on viral load, inclusion body formation, and host response in Pacific Herring with viral erythrocytic necrosis (VEN).

OBJECTIVE: The primary objective of this study was to determine the effects of temperature on viral erythrocytic necrosis (VEN) progression under controlled conditions. Secondarily, this study was intended to evaluate the combined effects of temperature and VEN on the Pacific Herring Clupea palasii transcriptome. METHODS: The effects of temperature on VEN progression were assessed by waterborne exposure of laboratory-reared, specific-pathogen-free Pacific Herring to tissues homogenates containing erythrocytic necrosis virus (ENV) at 6.9, 9.0, or 13.5°C. RESULT: Exposure of Pacific Herring to ENV resulted in the establishment of infections characterized by high infection prevalence (89%; 40/45) and mean viral loads (5.5 log10 [gene copies/µg genomic DNA]) in kidney tissues at 44 days postexposure. Mean viral loads were significantly higher in fish from the ambient (mean = 9.0°C) and warm (mean = 13.5°C) treatments (6.1-6.2 log10 [gene copies/total genomic DNA]) than in fish from the cool (mean = 6.9°C) treatment (4.3 log10 [gene copies/µg genomic DNA]). Similarly, the peak proportion of diseased fish was directly related to temperature, with cytoplasmic inclusion bodies detected in 21% of fish from the cool treatment, 52% of fish from the ambient treatment, and 60% of fish from the warm treatment. The mean VEN load in each fish (enumerated as the percentage of erythrocytes with cytoplasmic inclusions) at 44 days postexposure increased with temperature from 15% in the cool treatment to 36% in the ambient treatment and 32% in the warm treatment. Transcriptional analysis indicated that the number of differentially expressed genes among ENV-exposed Pacific Herring increased with temperature, time postexposure, and viral load. Correlation network analysis of transcriptomic data showed robust activation of interferon and viral immune responses in the hepatic tissue of infected individuals independent of other experimental variables. CONCLUSION: Results from this controlled laboratory study, combined with previous observations of natural epizootics in wild populations, support the conclusion that temperature is an important disease cofactor for VEN in Pacific Herring.

A geographic hot spot of Ichthyophonus infection in the southern Salish Sea, USA.

The prevalence of Ichthyophonus infection in Pacific herring Clupea pallasii was spatially heterogeneous in the southern Salish Sea, Washington State, USA. Over the course of 13 mo, 2232 Pacific herring were sampled from 38 midwater trawls throughout the region. Fork length was positively correlated with Ichthyophonus infection at all sites. After controlling for the positive relationship between host size and Ichthyophonus infection, the probability of infection was approximately 6-fold higher in North Hood Canal than in Puget Sound and the northern Straits (12 vs. 2% predicted probability for a 100 mm fish and 30 vs. 7% predicted probability for a 180 mm fish). Temporal changes in Ichthyophonus infection probability were explained by seasonal differences in fish length, owing to Pacific herring life history and movement patterns. Reasons for the spatial heterogeneity remain uncertain but may be associated with density-dependent factors inherent to the boom-bust cycles that commonly occur in clupeid populations.

Use of agent-based modelling to predict benefits of cleaner fish in controlling sea lice, Lepeophtheirus salmonis, infestations on farmed Atlantic salmon, Salmo salar L.

Sea lice, Lepeophtheirus salmonis, are ectoparasites of farmed and wild salmonids. Infestations can result in significant morbidity and mortality of hosts in addition to being costly to control. Integrated pest management programmes have been developed to manage infestations, and in some salmon farming areas, these programmes include the use of wrasse. Wrasse prey upon the parasitic life stages of L. salmonis and can be stocked on farms at varying densities. Despite considerable variation in the usage of wrasse, there are few quantitative estimates of how well they can control sea lice and how best to optimize their use. To explore at what densities wrasse should be stocked in order to meet specific control targets, we built an individual-based model that simulates sea lice infestation patterns on a representative salmonid host. Sea lice can be controlled through the use of chemical treatments as well as by wrasse predators. We found that the wrasse can effectively control sea lice, and the densities of wrasse needed for effective control depend upon the source of the infestation and the targeted level of control. Effective usage of wrasse can result in decreased use of chemical treatments and improved control of sea lice.

Identification of major water-soluble fluorescent components of some petrochemicals.

The chemical identities of several organic compounds that dominate the ultraviolet (UV) fluorescence of water after exposure to gasoline, diesel fuel and crude oil are presented. A combination of high-performance liquid chromatography with UV-fluorescence detection, fluorescence spectroscopy and gas chromatography-mass spectrometry (GC-MS) is used to show that naphthalene, methylnaphthalene and methylstyrene are the major fluorescent species in water following exposure to gasoline. These compounds are not dominant in water exposed to other petrochemicals we studied.

Calmodulin-antagonists inhibit vesicular Ca2+ uptake in Dictyostelium.

Chemotactic stimulation by cAMP elicits Ca2+ entry across the plasma membrane and uptake into intracellular Ca2+ stores. In order to better understand Ca2+ regulation in Dictyostelium we measured 45Ca2+ uptake in homogenates of aggregation competent cells. Besides the InsP3-sensitive store the acidosomes are responsible for Ca2+ transport. About 50% of the vesicular 45Ca2+ accumulation was inhibited by the calmodulin antagonist W-7 and 14% by the less efficacious analogue W-5. Half maximal inhibition by W-7 occurred at 37 microM concentration. Calmodulin antagonised the activity of W-7, and a monoclonal antibody against Dictyostelium calmodulin inhibited Ca2+ sequestration as did calmodulin antagonists of different classes. 100 microM BHQ-a SERCA-type Ca2+ transport ATPase blocker-inhibited most of the W-7 sensitive compartment and oxalate increased Ca2+ uptake into this compartment indicating that intracellular Ca2+ stores are the target of W-7. Ca2+/calmodulin thus seems to provide for a feedback regulation of Ca2+ sequestration.

On the confounding effects of phosphor persistence in oscilloscopic displays.

Phosphor persistence has been a source of confounding in studies of temporal integration in vision. We examined the confounding by assessing the effects of the persistence of two commonly-used phosphors (P15 and P31) on performance of a temporal-integration task. In one experiment we eliminated the visibility of phosphor persistence by closing two mechanical shutters upon display termination. In a second experiment we estimated the duration of phosphor persistence by displaying the image behind closed shutters which opened upon display termination. No detectable persistence was every produced by P15 phosphor. By contrast, P31 phosphor produced persistence that lasted several hundred milliseconds even when a veiling light was projected on the screen. We ascribe the earlier instances of confounding to inadequate interpretation of the technical data on phosphor decay.

Specific effects of the benzodiazepine midazolam on visual receptive fields in light and dark adapted human subjects.

Psychophysical experiments in humans have revealed similar characteristics of visual receptive fields as were found in cats and monkeys from retinal ganglion cell recordings. In addition, in some retinal ganglion cells of cats the GABA antagonist bicuculline decreases the activity of the inhibitory surround. These findings led to two predicitions: 1) benzodiazepines will selectively increase the inhibitory surround of human visual receptive fields, 2) after dark adaptation, no free GABA will be available in the synapses and benzodiazepines will have no effect on the visual system. Characteristics of human receptive fields were determined by subthreshold summation: the contrast threshold of a vertical line was measured dependent on the distance of two parallel flanking lines whose contrast was below threshold. Both hypotheses were confirmed: the threshold in the inhibitory region of receptive fields was specifically increased in a dose-dependent manner by midazolam PO (7.5 mg: P < 0.05; 15 mg: P < 0.01). In dark-adapted subjects no effect of midazolam was found. Control experiments with atropine (1 mg IV), sulpiride (100 mg IM), and levodopa (100 mg PO) showed no specific effect. The visual system may be a model to bridge the gap between animal and human psychopharmacology.

Visual masking: a reliable measure for the assessment of cognitive dysfunction in the elderly?

Reliable assessment of cognitive dysfunction in the elderly is a prerequisite for the evaluation of treatment of age-related cognitive decline. Psychophysical thresholds are known to be more reliable than psychometric tests, as assessed by stability of performance in visual masking. A backward (Till & Franklin, 1981) and a forward masking study (Coyne, 1981) were replicated. Thereafter, the same volunteers carried out a backward and forward masking task adapted to minimize noncognitive age-related influences: target and mask duration were individually adjusted to control for reduced eye transmissiveness. Attention was assessed with the electrooculogram; a dependent variable insensitive to sporadic decreases of attention was selected. Test-retest stability in the elderly after 2-4 weeks was r = .97 in backward and r = .86 in forward masking. As Alzheimer's disease mainly affects the cortex, backward masking, which is primarily cortical, may be useful to assess aspects of cognitive dysfunction.

Attention and eye movement control: an overview.

A review of recent experimental and theoretical findings is presented which relates the results of neurophysiological and psychological research on attention and eye movement to cognitive theorizing. With respect to the relation between shifts of attention and saccadic eye movements, several experiments have demonstrated that it is possible to dissociate the line of attention from the gaze direction. On the other hand, there is some indirect (although not yet conclusive) evidence that saccadic eye movements always produce a concomitant shift of attention. Another question is whether there is a moment-to-moment control of each consecutive saccade and latency or whether indirect control models (i.e. planning several saccades in advance) are more appropriate. The function of attentional processes which have to coordinate stimulus driven (bottom-up) and concept driven (top-down) mechanisms are discussed and related to some studies investigating local and global scanpaths.

A model of visible persistence and temporal integration.

A quantitative model of temporal integration and visible persistence is described and tested. The model treats visible persistence as resulting from processing activity within sustained visual channels whose temporal response is modelled using a second-order control system. Temporal integration of two successive stimuli is assumed to be enabled by the overlap between the two periods of activity and to be impaired by the non-overlapping activity. The model successfully predicts the effects of inter-stimulus interval and of stimulus duration on goodness of temporal integration.

Comparative study with enoxacin and netilmicin in a pharmacodynamic model to determine importance of ratio of antibiotic peak concentration to MIC for bactericidal activity and emergence of resistance.

An in vitro pharmacokinetic model was used to study the comparative antibacterial activities of multiple-dose regimens of enoxacin and netilmicin. Strains of Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, and Staphylococcus aureus were exposed to changing drug concentrations, mimicking human two-compartment pharmacokinetics. Oral administration was simulated for the quinolone, and intravenous administration was simulated for the aminoglycoside. Similar ratios of peak concentration to MIC resulted in similar changes in bacterial concentrations over time with both compounds. Following the initial dose, a rapid bactericidal effect occurred, with a greater than 99% reduction of the bacterial counts within 4 h at peak concentrations more than three times the MIC. However, bacterial regrowth occurred within 24 h unless the peak concentration/MIC ratio exceeded 8:1 (P less than 0.01). For the regrowing bacteria, MICs were four- to eightfold higher, and little or no bactericidal effect occurred following the second and subsequent doses. These data demonstrate the equally potent bactericidal activity of orally administered enoxacin and intravenously administered netilmicin. Selection of resistant subpopulations was similar with each drug. The peak concentration/MIC ratio may be an important parameter in the clinical use of quinolone and aminoglycoside antibiotics.

Impact of netilmicin regimens on the activities of ceftazidime-netilmicin combinations against Pseudomonas aeruginosa in an in vitro pharmacokinetic model.

The antibacterial activities of ceftazidime and netilmicin were studied in a two-compartment in vitro model. Pseudomonas aeruginosa cultures were exposed to changing drug concentrations that mimic human pharmacokinetics. Netilmicin alone reduced the numbers of organisms in cultures of the susceptible strains by more than 99% within 4 h; however, regrowth occurred after 8 h. Although ceftazidime alone killed more slowly than netilmicin, only one of the five strains regrew within 28 h. When both drugs were combined, rapid initial killing occurred without subsequent regrowth. Studied after 24 h in combination with ceftazidime, netilmicin was as effective when given as a single daily dose as when administered in three daily doses that provided 50% more aminoglycoside per day. Decreased bacterial susceptibility was seen after ceftazidime exposure for one strain and after netilmicin exposure for all originally netilmicin-susceptible strains. No such reduction in susceptibility was observed during exposure to the combination. The results of standard in vitro checkerboard tests for synergism were predictive of the initial (4 to 8 h) but not the final (24 to 28 h) assessment of drug interaction in the pharmacokinetic model.

Use of an in-vitro kinetic model to study antibiotic combinations.

A two compartment pharmacokinetic model was used to study combinations of piperacillin with N-formimidoyl thienamycin or amikacin, and azlocillin with netilmicin against strains of Pseudomonas aeruginosa. Antibiotic antagonism seen with in-vitro static tests of piperacillin and thienamycin did not occur with the kinetic model. Piperacillin plus amikacin showed enhanced activity, and azlocillin prevented bacterial regrowth seen with netilmicin alone during multiple dosing experiments at high bacterial inocula. This model is useful in the study of antibiotic combinations.

Beyond the displacement limit: an analysis of short-range processes in apparent motion.

If a circle of random dots is presented in two successive displays in which the second is rotated in relation to the first, then observers are able to accurately discriminate the direction of apparent rotation as long as the rotation is small. Rotations beyond this short-range apparent motion can produce the impression of motion in the reverse direction. The performance in identifying the direction of rotation further depends on the eccentricity of stimulation and the density of the random dots. Simulations of the experiments using the Marr and Ullman model of motion detection are in good quantitative agreement with the data except for low dot density patterns and large displacements. In these situations perception seems to be dominated by the operation of long-range processes.

Influence of diazepam and methylphenidate on identification of rapidly presented letter strings: diazepam enhances visual masking.

The effect of diazepam (0, 5, 10 mg) and methylphenidate (0, 5, 10 mg) on the identification of briefly displayed letter strings composing words was investigated. Each letter was presented for 70 ms. The interstimulus interval (ISI) between two successive letters was 10, 100 or 200 ms. The length of the words (WL) was three, five, seven or nine letters. The dependent variable was the number of correctly identified words. As predicted, strong visual masking effects, the strength of which was dependent on ISI and WL, appeared. In the 10 ms condition, diazepam at both doses, but not methylphenidate, further enhanced masking. It is suggested that the results reflect the effect of GABA on masking, and support certain psychological theories about the inhibitory neuronal pathways that are involved in masking. The human peripheral visual system is a promising model for the experimental assessment and integration of pharmacodynamic, neurophysiological and psychological observations.