Chronotoxicology of florfenicol

Sixty 3-month-old homozygote male mice were studied for circadian rhythmicity in the toxicity of florfenicol overdose. Animals were kept under a regimen of 12h light, 12h darkness (12:12 LD) with food and water available ad libitum. The LD50 (median lethal) dose was determined in a preliminary experiment and was administered to groups of 10 mice at six different clock times (hours) after light onset (HALO): 0, 4, 8, 12, 16, and 20 HALO. Cosinor analysis verified a statistically significant (P < .04) circadian rhythm in the toxic effect (mortality) of florfenicol. Mortality was greatest when the drug was injected 4h after the commencement of the activity span (16 HALO) and least when injected 4h after the start of the diurnal rest span (4 HALO). Mortality was 2.5 times greater when drug injection was given at 16 HALO than at 4 HALO.     

Circadian rhythm of physiological color change in the amphibian Bufo ictericus under different photoperiods

Ectothermic vertebrates can exhibit chromatic adaptation to the environment. The aim of this work was to characterize the rhythm of color change of the amphibian Bufo ictericus, submitted to different photoperiodic regimens, as quantified by skin reflectance values. Adult males were maintained under a 12:12 Light/Dark (LD) cycle during seven days before every experiment. During the experiments, animals were kept in individual boxes for 8 days, under the following photoperiodic regimens: LD 12:12, LD 14:10, DD and LL. In the last 3 days of the treatments, the reflectance of the toad dorsal skins was measured at 3-h intervals, with the aid of a reflectometer. A 3-day time series consisting of 8 data points per day was obtained, which was analyzed by the Cosinor method. The analysis demonstrated that the reflectance values exhibited significant circadian oscillations in the regimens LD 12:12, LD 14:10 and DD, suggesting that the specie B. ictericus shows an effective circadian rhythm of color change. The reflectance values did not exhibit a significant circadian rhythm in the LL regimen showing that this is a condition not permissive for the expression of the color change rhythm.     

Chronosynergistic effects of lighting schedule-shift and cefodizime on plasmacytoma growth and host survival time

Lighting regimen shifts can modify the effects of cefodizime, for the purpose of a chronoimmunomodulation. Two experiments were carried out on male and female LOU rats inoculated subcutaneously with plasmacytoma cells. Some rats were kept on their original LD12:12 regimen, whereas others, after tumor implantation, were subjected every second day to 6-h shifts, instituted, in alternation, as advance or delay. Daily treatment with cefodizime or placebo started when, overall, about 50% of the animals had developed a palpable tumor. A subgroup of animals contributed daily smears for the determination of the estrus cycle and further provided core temperature and activity data by telemetry. In Experiment I, the repeated shifting of the LD regimen was associated with survival time prolongation (p less than 0.05), irrespective of drug administration. Moreover, in those (female) rats repeatedly exposed to shifts of the lighting schedule, cefodizime was found to prolong survival time (p less than 0.05). The effects of cefodizime vs placebo on survival time were found to be circadian stage-dependent. In Experiment II, differing from Experiment I in the initial conditions before the institution of the shifts, cefodizime treatment was associated with a prolongation of survival time of the female rats kept on a fixed LD12:12 regimen. Both male and female rats again showed a circadian stage-dependence of the cefodizime effect. These results suggest that interactions between synchronizers of rhythms (such as shifts of the lighting regimen, the latter simulating the daily routine) and immunomodulating agents such as cefodizime may be optimized to improve treatment strategies against cancer and other diseases.     

EXPERIMENTS ON THE MODE OF ACTION OF DIELDRIN SEED-DRESSINGS AGAINST SEEDLING ATTACK BY THE LARVAL FRIT FLY, OSCINELLA FRIT L

Experiments with dieldrin seed-dressings were done to see whether the conditions favouring their action against wheat bulb fly larvae apply also to frit fly.
Experiments with oats and wheat in boxes and a small field trial with oats showed that fewer newly hatched larvae were killed when dieldrin-treated seed was sown deeply than when sown 1/4 - 1/2 in. deep; shallow sowing probably puts the insecticide where the larvae are likely to meet it before they attack the plant. Kill was never complete even with dieldrin applied at almost forty times more than the highest rate normally used. This is partly because some larvae hatch from eggs on the plant and these, unlike those from soil eggs, can enter the shoot without touching the seed dressing.
Even in the most favourable circumstances (large doses of dieldrin applied to the shallow-sown seed and the plant at the one-leaf stage at the time of attack) there was little evidence that frit fly larvae are killed inside the shoot by systemic action of dieldrin translocated from the seed. This is not because the pales which normally surround the oat seed impair the uptake of insecticide, or because of any other factor specific to oats. Frit fly larvae also survived in wheat from seed dressed with doses of dieldrin that can kill almost all wheat bulb fly larvae by systemic action. Possible reasons for this difference are discussed.     

The Comparative Susceptibility of the Diamondback Moth Plutella xylostella and Some Other Major Lepidopteran Pests of Brassica Crops to a Range of Baculoviruses

The susceptibility of larvae of the Diamondback moth (DBM), Plutella xylostella to infection by three baculoviruses was evaluated in the laboratory using a microdroplet feeding assay. The viruses tested were a granulovirus (GV), originally isolated in Taiwan from P. xylostella larvae (Px GV-Taiwan); the nucleopolyhedrovirus (NPV) from Galleria mellonella (Gm NPV), and the NPV from Autographa californica (Ac NPV). Neonate P. xylostella larvae were susceptible to infection by all three viruses. In an extensive series of bioassays carried out over a 21-month period, LD 50S for neonate DBM larvae ranged from 1.0-8.9 viral occlusion bodies (OB) for Px GV-Taiwan, and 9.5-30.2 OB for Gm NPV and Ac NPV. LT 50S for the three viruses ranged from 3.8-6.0 days at 27 C, with Gm NPV having a significantly shorter LT 50 than the other two viruses. Second and third instar larvae of P. xylostella were significantly less susceptible to infection by Px GV-Taiwan (LD 50 s ranging from 18-57 OB/larva) than were neonate larvae. Gm NPV also initiated infection in several other lepidopterous pest species that colonize brassica crops. In particular, neonate Crocidolomia binotalis larvae proved highly susceptible to Gm NPV, with mean LD 50 s ranging from 2.1 to 9.3 OB/larva and a mean LT 50 of 4.8 days at a dose of 8.08 OB. Heliothis virescens neonate larvae were also highly susceptible to Gm NPV (LD 50 , 7.1 OB), but Mamestra brassicae larvae were less so (LD 50 , 80-270 OB). The results of the bioassays suggest that Px GV-Taiwan is highly infective and could be developed as a selective microbial pesticide for DBM. While Gm NPV has a higher LD 50 in DBM larvae, its wider host range may be of considerable value in situations where DBM occurs on cruciferous crops together with a complex of other lepidopterous pests.     

Circadian Variation in Human Performance Evaluated By the Walter Reed Performance

-As part of a two clock-time (0830 versus 2030) evaluation of administration-time dependent effects of dexedrine (S mg) and triazolam (0.25 mg) on human cognitive performance, placebo (control) studies were conducted on 12 diurnally active (0700-2300) male adults (23-38 yrs) using a double-blind, randomized crossover design. Testing was conducted hourly during a series of sleepless 13-hr spans commencing in the morning or evening, using the Walter Reed computer controlled and scored multi-task cognitive performance assessment battery. For the placebo condition, Single and Group Cosinor analyses documented circadian rhythms in performance for most tasks (reaction time, logical reasoning, serial add/subtract and spatial orientation) both for individuals and the group. Overall, performance was worse overnight, when sleepiness was greatest, and best between 1830 and 2030. It was most variable around 0600-0700. The day-night variation in performance over all cognitive tests amounted to 21% of the 24-hr mean.     

CHRONOTOXICOLOGY OF FLORFENICOL

Sixty 3-month-old homozygote male mice were studied for circadian rhythmicity in the toxicity of florfenicol overdose. Animals were kept under a regimen of 12h light, 12h darkness (12:12 LD) with food and water available ad libitum. The LD₅₀ (median lethal) dose was determined in a preliminary experiment and was administered to groups of 10 mice at six different clock times (hours) after light onset (HALO): 0, 4, 8, 12, 16, and 20 HALO. Cosinor analysis verified a statistically significant (P <. 04) circadian rhythm in the toxic effect (mortality) of florfenicol. Mortality was greatest when the drug was injected 4h after the commencement of the activity span (16 HALO) and least when injected 4h after the start of the diurnal rest span (4 HALO). Mortality was 2.5 times greater when drug injection was given at 16 HALO than at 4 HALO. (Chronobiology International, 18(3), 567–572, 2001)     

Persistence and distribution of insecticidal seed-dressings for control of the wheat bulb fly, Leptohylemyia coarctata

Samples of plants and soil from two experimental sites, one clay loam and one peaty loam, were analysed chemically to try to explain differing results with γ-BHC and dieldrin seed-dressings applied to control wheat bulb fly on different soil types, and to suggest reasons for patchy plant stands and occasional failure to control the pest. Seed dressed with dieldrin yielded more than untreated seed on both sites, whereas seed treated with γ-BHC yielded as much as seed treated with dieldrin on the clay site, but it yielded less than untreated seed on peat. The chemical analyses showed that the poor performance of γ-BHC on the peat site could not be explained by its failure to persist, because the soil still contained considerable quantities of γ-BHC at the time of insect attack. Possible reasons may be sorption of the poison by organic matter making it less available, and deeper sowing, permitting larvae to reach the plants without prolonged contact with the insecticide. Bayer 38156 (O-ethyl S-p-tolyl ethyl phosphonodithioate) and trichloro-nate (O-ethyl O-2,4,5-trichlorophenyl ethyl phosphonothionate) persisted in soil long enough and were sufficiently toxic to wheat bulb fly to suggest that organophosphorus compounds might provide alternatives to chlorinated hydrocarbon insecticides for control of the fly. Analyses made on seeds dressed at the laboratory for the experiment showed that the amounts of insecticide on seeds were smaller than expected and that the amounts on individual seeds differed greatly. Of ten samples of seeds dressed commercially with y-BHC, three carried very little insecticide, and the variations in the other seven samples were greater than with experimentally applied dressings.     

Circadian Variation in Human Performance Evaluated By the Walter Reed Performance

–As part of a two clock-time (0830 versus 2030) evaluation of administration-time dependent effects of dexedrine (S mg) and triazolam (0.25 mg) on human cognitive performance, placebo (control) studies were conducted on 12 diurnally active (0700–2300) male adults (23–38 yrs) using a double-blind, randomized crossover design. Testing was conducted hourly during a series of sleepless 13-hr spans commencing in the morning or evening, using the Walter Reed computer controlled and scored multi-task cognitive performance assessment battery. For the placebo condition, Single and Group Cosinor analyses documented circadian rhythms in performance for most tasks (reaction time, logical reasoning, serial add/subtract and spatial orientation) both for individuals and the group. Overall, performance was worse overnight, when sleepiness was greatest, and best between 1830 and 2030. It was most variable around 0600–0700. The day-night variation in performance over all cognitive tests amounted to 21% of the 24-hr mean.     

Behaviour and fitness of γHCH/dieldrin resistant and susceptible female Anopheles gambiae and An.stephensi mosquitoes in the absence of insecticide

The effects of gamma HCH/dieldrin resistance genes on various fitness components of mosquito larvae and adult females in the absence of insecticide were investigated in backcrossed strains of Anopheles gambiae Giles and An.stephensi Liston. Among larvae, heterozygotes (RS) developed slightly but significantly faster than homozygotes for resistance (RR) or susceptibility (SS). The lifetime fecundity of RR females in population cages was only half to two-thirds that of SS and RS females despite similar longevities; several reasons were identified: RR gravid females were less responsive to oviposition-site stimuli, their spontaneous activity--as measured in an acoustic actograph--was only half that of SS or RS females, and RR females produced fewer eggs per unit bloodmeal. When inseminated females were recorded in LD 12:12, RR were again less active than SS or RS. When the lighting was switched to a regime simulating full-moonlight, the activity pattern of SS and RS changed and they flew for longer periods. In contrast, the activity of RR females was the same in LD 12:12 as in 'moonlight'. In a test simulation of potential predation, RR mosquitoes took to flight least readily. All component tests on adult females therefore point to RR as being the least fit of the three genotypes. The behavioural tests suggest that resistance has raised the response threshold of RR females to diverse stimuli. A possible physiological mechanism underlying RR behaviour is that a change in the cyclodiene receptor on the chloride channels has increased their permeability to chloride ions, causing hyper-inhibition of the nervous system.     

Biological rhythm of leukocyte concentration in calves under natural conditions

The objective of this study was to describe daily rhythms in leukocyte concentration in calves. The work was carried out during June - July 2004. Six animals were exposed to a natural photoperiod of about 10:14 LD. 5 ml blood samples were drawn from the jugular vein every two hours during the 1st, 4th, 14th, 20th and 30th days of life. Concentration of leukocytes was measured with a hematological counter and analyzed with repeated measures ANOVA and Cosinor analysis. Leukocyte concentration did not show a circadian rhythm at the moment of birth, but exhibited ultradian components that adjusted to daily patterns by the 30th day of life, with an acrophase at 0:35 h and R = 0.97%.     

Cross-resistance between dieldrin and fipronil in German cockroach (Dictyoptera: Blattellidae)

The toxicity of fipronil and dieldrin was determined in one susceptible laboratory strain and seven insecticide-resistant field-collected strains of Blattella germanica (L). The Zo960302 and Ga021001 strains were 1,270- and 2,030-fold resistant to dieldrin and 15- and 14-fold resistant to fipronil. The Su960304 and Od010803 strains were 15- and 13-fold resistant to dieldrin and two- and four-fold resistant to fipronil. Three strains showed no or a low level of resistance to dieldrin and fipronil. Crosses were performed between the susceptible strain Danish Pest Infestation Laboratory (DPIL)-SUS and the resistant strains Zo960302 and Su960304 and resistance to dieldrin and fipronil were intermediate compared with the susceptible and the resistant strains. Backcrosses to both of the parental strains showed cosegregation of dieldrin and fipronil resistance. The toxicity of dieldrin and fipronil was correlated when compared at LD50, and 93% of the observed variation in LD50 of fipronil can be ascribed to variation among predictions based on the value of LD50 of dieldrin. The frequency of the A302S substitution in the resistance to dieldrin (Rdl) gene in the highly dieldrin- and fipronil-resistant strains Zo960302 and Ga021001 and the moderately resistant Su960304 was 0.97, 1.0, and 0.38, respectively. We consider the connection between the frequency of the Rdl mutation and dieldrin and fipronil resistance a causal connection and not merely a coincidence.     

Perspectives in chronobiology of air pollution

In a series of experiments, male and female Sprague Dawley rats, kept in light (L) from 06(00) to 18(00) alternating with darkness (LD 12:12) inhaled different concentrations of carbon monoxide (50-1,700 ppm) at each of two test times, 12 h apart. A decrease in flow of CO2 (VCO2) resulting from CO inhalation was greater in the active dark (D) than resting light (L) span. Experimental hypoxic mortality of male and female mice also shows circadian variations, being greater in the D than in the L span. Moreover, a difference of mortality was observed betwen hypoxic exposures performed at 12(00) (in LD or DL) and hypoxic exposures performed at 00(00) (in LD or DL). Such results await tests of any extent to which they model responses of human beings to air pollution. In human beings any external environmental circadian, circaseptan and circannual variations in air pollution as such may serve to variable extent as socioeconomic synchronizers of innate rhythms with a corresponding frequency, rather than as solely generators of time patterns in any physiopathologic response to air pollution.     

Botanical insecticides for controlling agricultural pests: piperamides and the Colorado Potato Beetle Leptinotarsa decemlineata say (Coleoptera: Chrysomelidae)

The efficacy of extracts from two Piperaceae species, Piper nigrum L. and P. tuberculatum Jacq. were evaluated using larvae and adults of the Colorado Potato Beetle Leptinotarsa decemlineata (Say). Young larvae and neonates were the most susceptible; a 24-h LD(50) of 0.064% extract of P. tuberculatum was determined for 4-day-old larvae, while 0.05% extract of P. nigrum reduced larval survival up to 70% within one week after treatment of potato Solanum tuberosum L. (Solanaceae) plants. When an insecticide resistant strain of L. decemlineata larvae was tested with the P. tuberculatum extract, there was less than a 2-fold tolerance ratio compared to the 22-fold tolerance ratio to cypermethrin, a pyrethroid. Older larvae, pre-pupal stage and adults, were less sensitive to the P. nigrum extracts; the 24-h LD(50) was 0.5% (95% C.I. = 0.36, 0.65). However, the same concentration was equally effective under field conditions. In the greenhouse, P. nigrum at 0.5% was as effective at reducing adult L. decemlineata feeding as combinations with 2 separate botanical mixtures, garlic and lemon grass oil. Under field conditions, the residual activity of the P. nigrum extracts was less than 3 h. When adult L. decemlineata were placed on treated plants exposed to full sunlight for 0, 1.5, and 3 h, leaf damage progressively increased as the main active compound, piperine, was found to degrade by 80% after 3 h. An in vitro polysubstrate monoxygenase (PSMO) enzyme assay, using the substrate methoxyresorufin O-demethylation (MROD), determined that the principal P. nigrum active compound, piperine, is responsible for inhibition of that specific enzyme. The results suggest that Piper extracts could be used effectively as contact botanical insect control agents to protect potato plants from developing L. decemlineata larvae at concentrations less than 0.1%. There is also potential for Piper extracts to control insecticide resistant populations in conjunction with other integrated pest management (IPM) strategies used in conventional and organic agriculture.     

Effects of several insecticide formulations on oothecal drop and hatchability in German cockroaches (Dictyoptera: Blattellidae)

Formulations used for control of German cockroaches, Blattella germanica (L.), were investigated. The insecticides tested on gravid females were bendiocarb, chlorpyrifos, cyfluthrin, cypermethrin, fenvalerate, hydramethylnon, malathion, propetamphos, propoxur, and pyrethrins. Exposure to each insecticide increased the frequency of oothecal drop and reduced the percentage oothecal hatch. The percentage of oothecae that were dropped increased as a curvilinear function of insecticide concentration. An LD50 of propoxur to female German cockroaches resulted in the greatest oothecal drop (83.50%); fenvalerate caused the least drop (22.60%). LD50's of cypermethrin, propetamphos, and propoxur resulted in 29.60, 31.807, and 37.30% hatch from dropped oothecae, respectively. The smallest percentage hatch from retained oothecae was caused by LD50's of propoxur (1.5%) and cyfluthrin (7.70%). Retained oothecae from females treated with an LD50 of fenvalerate (68.70%) or pyrethrins (68.70%) had the greatest percentage hatch. Total percentage hatch (dropped and retained oothecae) declined exponentially as the insecticide concentration increased. An LD50 of cypermethrin limited total oothecal hatch to 24.50%, whereas LD50's of fenvalerate, malathion, and pyrethrins resulted in 53.60, 52.20, and 58.90% hatch, respectively. Mean time to oothecal hatch increased linearly with increasing insecticide concentration for all insecticides tested.     

Social Cues Accelerate Reentrainment of Circadian Rhythms in Diurnal Female Octodon Degus (Rodentia-Octodontidae)

Previous studies paired diurnal Octodon degus undergoing/phase advances (phase-shifters) with those entrained to a light-dark (LD) cycle (donors). Results included opposite outcomes of male and female social cues on resynchronization following 6-h advances in females, but no effect of social cues on male resynchronization. The first experiment determined if social cues could influence resynchronization rates of circadian rhythms in male and female degus following a 6-h phase delay of the LD cycle. Female phase-shifters resynchronized temperature and activity rhythms 20-35% faster when housed with either entrained (donor) females or males compared with females housed alone. No significant differences in resynchronization rate for phase-shifting males existed between test conditions. This experiment extends the previous finding that females, but not males, respond strongly to donor cues to increase resynchronization rates in the presence of light. A second experiment determined that accelerated resynchronization rates of female phase-shifters housed with female donors were due to social cues directly affecting the circadian system rather than the result of social masking. On the day following resynchronization with or without a female donor present, phaseshifters were transferred individually to constant conditions (DD). The temperature and activity rhythms of female phase-shifters free-ran from the point at which resynchronization occurred for both the control and experimental females. Thus, social cues accelerate true reentrainment, not masking, of the circadian system in the presence of a LD cycle in female degus. Donor cues from females enhance reentrainment after advances and delays, but the effect of male donor cues is dependent on the direction of the phase shift.     

Genetic linkage between malathion and dieldrin resistance in Anopheles arabiensis

A strain of Anopheles arabiensis resistant to both malathion and dieldrin was crossed and backcrossed to a susceptible strain. The progeny were tested on each insecticide in turn. Less than 50% mortality in the second insecticide exposure among the backcross progeny indicated linkage between the resistance genes. In a backcross of A. gambiae X A. arabiensis hybrids a recombination rate of 7.5% was observed. A Y-translocation strain of A. arabiensis showed less than 2.8% recombination between the resistance genes. It is impossible to confirm the genotype of apparent recombinants using existing stocks, but the two resistance mechanisms are biochemically distinguishable. If the two genes are very closely linked, linkage disequilibrium could influence the consequences of switching to malathion spraying after dieldrin resistance has evolved.     

Light regulates the cell cycle in zebrafish

The timing of cell proliferation is a key factor contributing to the regulation of normal growth. Daily rhythms of cell cycle progression have been documented in a wide range of organisms. However, little is known about how environmental, humoral, and cell-autonomous factors contribute to these rhythms. Here, we demonstrate that light plays a key role in cell cycle regulation in the zebrafish. Exposure of larvae to light-dark (LD) cycles causes a range of different cell types to enter S phase predominantly at the end of the day. When larvae are raised in constant darkness (DD), a low level of arrhythmic S phase is observed. In addition, light-entrained cell cycle rhythms persist for several days after transfer to DD, both observations pointing to the involvement of the circadian clock. We show that the number of LD cycles experienced is essential for establishing this rhythm during larval development. Furthermore, we reveal that the same phenomenon exists in a zebrafish cell line. This represents the first example of a vertebrate cell culture system where circadian rhythms of the cell cycle are observed. Thus, we implicate the cell-autonomous circadian clock in the regulation of the vertebrate cell cycle by light.     

Circadian and Seasonal Variations of Plasma Insulin and Cortisol Concentrations in the Syrian Hamster, Mesocricetus Auratus

Circadian rhythms of plasma insulin, Cortisol, and glucose concentrations were examined in scotosensitive (reproductively sensitive to inhibitory effects of short daylengths) and scotorefractory male and female Syrian hamsters (Mesocricetus auratus) maintained on short (LD 10:14) and long (LD 14:10) daylengths. The baseline concentration (mean of all values obtained every 4 hr six times of day) of insulin was much greater in female than in male scotosensitive hamsters kept on short daylengths. These differences in insulin concentration may account for the observed heavy fat stores in female and low fat stores in male scotosensitive hamsters kept on short daylengths. The baseline concentrations of Cortisol were approximately equal in both scotosensitive and scotorefractory males held on short and long daylengths, but were relatively low in females held on short daylengths and especially high in scotorefractory females held on long daylengths.

The plasma concentrations of both cortisol and insulin varied throughout the day in many of the groups tested. However, the variations were not equivalent. The circadian variations of cortisol were similar irrespective of sex, seasonal condition and daylength. Peak concentrations generally occurred about 12 hr after light onset. In contrast, the circadian variations of insulin differed markedly. For example in male hamsters, robust daily variations were found in scotosensitive hamsters held on short daylengths but not on long daylengths and in scotorefractory hamsters held on long daylengths but not on short daylengths. Furthermore, the daily peak occurred during the light in the scotosensitive hamsters and during the dark in the scotorefractory animals. Neither the daily feeding pattern (about 60% consumed during dark) nor the daily variations of glucose concentration varied appreciably with seasonal condition or daylength. They do not appear to determine nor directly reflect the variations in cortisol and glucose concentrations. It is postulated that the daily rhythms of cortisol and insulin are regulated by different neural pacemaker systems and that changes in the phase relations of circadian systems account in part for seasonal changes in body fat stores.