Pinealectomy Does Not Modulate the Characteristics of 24-h Variation in Air-Gulping Activity of Clarias batrachus

The effect of pinealectomy on the characteristics of circadian variation in air-gulping activity of Clarias batrachus was examined. Live Clarias batrachus of mixed sex (40-50 gm body weight) were procured from the local market and kept in the stock aquaria under the laboratory conditions for proper acclimation. Following 7 days of acclimation, animals were randomly selected from the stock aquaria and divided into two groups. The pineal was surgically removed from fishes in the first group (P×G) and the second group was left intact (IG). The individuals were kept in the specially designed glass aquaria. They were exposed under LD 12:12 (Lights on at 06:00) inside the chronocubicles for recording of the air-gulping activity. Results clearly indicate that the air-gulping activity exhibits a significant 24 h rhythm in both Intact (IG) and Pinealectomized (P×G) groups of this catfish. The peak of the air-gulping activity rhythm was located in the dark phase of the LD cycle in both groups. It may be concluded that pinealectomy does not modulate the 24 h rhythm in air-gulping activity of Clarias batrachus under LD 12:12.     

Effect of Pinealectomy on Re-entrainment of Air-gulping Activity in Indian Walking Catfish, Clarias batrachus

The present study was aimed to examine the effect of pinealectomy on re-entrainment of air-gulping activity in Indian walking catfish Clarias batrachus under various LD phases. Live Clarias batrachus of mixed sex (40-50g body weight) were procured from the local market and kept in the stock aquaria under the laboratory conditions for proper acclimation. Following 7 days of acclimation, animals were randomly selected from the stock aquaria and divided into 2 groups. The pineal was surgically removed from one group (P_xG) and the other group was treated as intact control group (IG). Both groups were exposed under LD 12:12 cycle inside the chronocubicles. In every 7 days LD schedule was delayed by 4 hours by lengthening of the light-on time. Both intact and pinealectomized animals are exhibited 24h rhythm in air-gulping behaviour and its entrainability irrespective of phase shifts of LD cycle. However, the phase angle of peak with reference to light on hour gradually decreased from phase 1 to phase 4, irrespective of treatment (intact/pinealectomized). Further, it has been noticed that the occurrence of multifrequency rhythm (24h, 12h and 6h) is being decreased from phase 1 to phase 4. It could be concluded that daily light-dark cycle plays an important role in modulating the rhythmic characteristics of air-breathing activity in Clarias batrachus and that pinealectomy does not modulate the effects of LD cycles on such activity.     

Circadian Variation in Phototactic Behaviour of Walking Indian Catfish, Clarias batrachus

Temporal organization in phototactic rhythm in male Clarias batrachus was studied in a choice chamber consisting of a photic zone and an aphotic zone. Results indicate that most of the fishes exhibited a statistically significant 24-h variation in the frequency of visits to the photic zone with a peak frequency at late night or early morning. Cosinor analysis of phototactic index (PI) values also yielded comparable results. This behaviour of C. batrachus shows similarity with its other activities, like air gulping, surfacing, foraging or predation that occur chiefly at night. The light intensity at the surface of a natural water body is more than at the bottom, even at night. Therefore, the peak phototactic activity observed in C. batrachus in the night or early morning could be attributed to its daily migration from the bottom to the surface of the water bodies. The results of the present study might be useful in aquaculture practice and inland fishing.     

Does starvation influence the antioxidant status of the digestive gland of Nacella concinna in experimental conditions?

In a previous study we analysed the effect of diesel seawater contamination in the digestive gland of the Antarctic limpet Nacella concinna. We observed that antioxidant enzyme activities decreased after one-week starvation prior to the experiment, and this was considered in the analysis of the obtained results. To know whether the digestive gland oxidant-antioxidant status may be altered by starvation and experimental conditions, we evaluated the food deprivation effect in limpets from the nearshore shallow waters of Potter Cove, Antarctica. Organisms were acclimated to laboratory conditions and were divided in fed and starved groups, and maintained in these conditions during one month. Every week 20 limpets were sampled from each group. Digestive glands were dissected and kept frozen until they were processed. Superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST) activities, as well as lipid peroxidation (LPO) measured as thiobarbituric reactive substances (TBARS), protein oxidation (PO) and reduced glutathione (GSH) were measured. For both groups of limpets, SOD increased its activity in the first week of the exposure period, with a maximum in the second week. CAT activity increased significantly in the second week, only for the starved group. Similarly, GST activity also increased for starved group in the second week; but maintained this tendency for both groups until the fourth week. In fed and starved limpets, TBARS values increased significantly, during the first week and then returned to normal values. The PO levels in the starved group increased only during the first week. The GSH content, for the fed group, increased significantly after the third week. The obtained results indicate that biochemical or physiological studies conducted with N. concinna should consider the effects of food deprivation and time spent under experimental conditions.     

Effect of short light-dark cycles on young and adult TGR(mREN2)27 rats

Animals placed under short light-dark (LD) cycles show a dissociation of their circadian rhythms. However, this effect has only been studied in Wistar rats and with the motor activity (MA) rhythm. Thus, in the present experiment, we studied in TGR(mREN2)27 (TGR) rats, a strain of hypertensive rats, the effect of a short LD cycle on the circadian rhythms of MA, heart rate (HR), and blood pressure (BP). Our aim was [1] to investigate whether the exposure of TGR rats to a short LD cycle induced a dissociation of their circadian rhythms, [2] to study the effect of short LD cycles on the development of the circadian rhythms of TGR rats, and [3] to compare the effect of short LD cycles on young and adult TGR rats. One group of TGR rats was maintained under LD cycles of 22h periods (group G22). The progress in time of their rhythms was compared to that of TGR rats of the same age that had been kept under LD cycles of 24h periods (group G24). For the third point, the rhythms of a group of 5-week-old TGR rats kept under LD 22h cycles (young rats) were compared to those of a group of 11-week-old TGR rats (adult rats). Results showed that there is a dissociation of the circadian rhythms of all the variables monitored in TGR rats maintained under LD 22h cycles, independent of age. We have also found that group G22 showed a higher increase in BP with age and a higher mortality due to malignant hypertension compared to group G24. Finally, it seems that it is harder for young rats to entrain to short LD cycles than for adult rats, and young rats have a higher mortality due to malignant hypertension than adult rats. In conclusion, we demonstrated that short LD cycles produce a dissociation in the HR, BP, and MA circadian rhythms. The results of this experiment, compared to those previously obtained in Wistar rats, suggest that the light perception, the responses of the circadian system to light, or both are altered in the TGR rats. (Chronobiology International, 18(4), 641-656, 2001)     

Pinealectomy Does Not Modulate the Characteristics of 24-h Variation in Air-Gulping Activity of Clarias batrachus

The effect of pinealectomy on the characteristics of circadian variation in air-gulping activity of Clarias batrachus was examined. Live Clarias batrachus of mixed sex (40–50 gm body weight) were procured from the local market and kept in the stock aquaria under the laboratory conditions for proper acclimation. Following 7 days of acclimation, animals were randomly selected from the stock aquaria and divided into two groups. The pineal was surgically removed from fishes in the first group (P×G) and the second group was left intact (IG). The individuals were kept in the specially designed glass aquaria. They were exposed under LD 12:12 (Lights on at 06:00) inside the chronocubicles for recording of the air-gulping activity. Results clearly indicate that the air-gulping activity exhibits a significant 24 h rhythm in both Intact (IG) and Pinealectomized (P×G) groups of this catfish. The peak of the air-gulping activity rhythm was located in the dark phase of the LD cycle in both groups. It may be concluded that pinealectomy does not modulate the 24 h rhythm in air-gulping activity of Clarias batrachus under LD 12:12.     

Life history response and age-specific tolerance to starvation in Brachionus plicatilis O.F. Müller (Rotifera)

To examine the life history response and age-specific tolerance to starvation in the rotifer Brachionus plicatilis O.F. Müller, we carried out two series of individual culture experiments. In the first experiment, rotifers were fed until each of the ages of 1-4 days, and were then starved during the rest of their lifetimes. The control group was fed throughout their lifespans. Rotifers stopped active reproduction just after the onset of food deprivation, and showed shorter subsequent survival times when they were starved at older ages. The finding that the larger the number of offspring produced before food deprivation, the shorter the subsequent lifetime under starvation, appeared to reflect a trade-off with the cost of reproduction. In the second experiment, newborns were starved until each of the ages of 1-5 days, and were fed thereafter. The lifespans of the rotifers starved up to the age of 3 days were not statistically different from those that were not starved. Although the starved rotifers began to reproduce once fed again, their lifetime fecundity decreased significantly from that of the non-starved group. Based on these results, it was suggested that the reproductive suppression caused by starvation would cause rotifers to have a longer lifespan to allow for future reproduction.     

Effects of starvation and refeeding on elastase-induced emphysema

Adult rats received pancreatic elastase (75 U/100 g) intratracheally and were divided into three groups: fed, starved, and refed. Starved rats received one-third of their measured daily food consumption until they lost 40% body weight. The refed group was fed after 40% weight loss. A control group received saline intratracheally. Saline volume-pressure curve was shifted more significantly to the left of the control group in starved than in fed rats and was superimposed in refed and fed groups. Mean linear intercept was larger and alveolar surface area was smaller in starved than in fed rats compared with the control group; both were similar in fed and refed rats. Protein and hydroxyproline content of the lung were higher in fed than in control and in starved groups; after refeeding these returned to the control values. We conclude that starvation aggravates elastase-induced injury and that refeeding results in the complete recovery of the mechanical but only partial recovery of the morphometric changes induced by starvation.     

Liver arginase activity and urinary nitrogen products profile in adult Bufo arenarum under different protein intake

The effect of different levels of diet protein on adult Bufo arenarum liver arginase activity and protein content, plasma urea and urinary profile of nitrogen waste products was estimated. Animals kept under environmental constant conditions were submitted to a nutritional standardization period being fed beef meat daily during four days. Then animals were distributed in three groups: Group 0 (control), that was sampled at the end of the standardization period; Group 1, that was starved for 18 days and Group 2, that was fed daily for 18 days and then sampled. With respect to controls, liver arginase specific activity was significantly lower in starving toads (Group 1); liver protein content was elevated in fasted animals (Group 1) and plasma urea concentration increased in the intensive feeding group (Group 2). Urinary nitrogen end products in animals from both control and experimental groups showed no changes either in their absolute values or in their partition percentage rates.     

Circadian Variation in Circulating Levels of Melatonin, Testosterone and Estradiol-17β in a Tropical Freshwater Catfish, Clarias batrachus, during Different Phases of the Annual Reproductive Cycle

In the present investigation, daily variations in plasma levels of melatonin, testosterone and estradiol-17β were analysed in the tropical freshwater catfish, Clarias batrachus during early and late-preparatory, pre-spawning, late-spawning and post-spawning phases of its annual reproductive cycle. Plasma melatonin levels exhibited a significant circadian rhythm during all the reproductive phases but peaks were invariably at night. The amplitude of melatonin was high during the early-preparatory phase and low in the pre-spawning phase. Testosterone also displayed significant rhythm during all the studied phases except post spawning phase. Estradiol-17β revealed diurnal rhythm only during preparatory and pre-spawning phases. The peak in the levels of plasma testosterone and estradiol-17β were recorded during the photophase. During the late-preparatory phase, the rhythm of testosterone coincided with the rhythm of estradiol-17β.     

Effect of anthropogenic feeding regimes on activity rhythms of laboratory mussels exposed to natural light

Anthropogenic disturbance may affect animal behaviour and should generally be minimised. We examined how anthropogenic disturbance (24 h food deprivation) affected circadian rhythms in laboratory mussels Mytilus edulis exposed to natural light in the absence of tides. Repeated measures data were collected on mussel gape angle, exhalant pumping and valve adduction using a Hall sensor system over eight consecutive 24 h periods when exposed to two feeding conditions after 24 h food deprivation. Mussels (fed once per day at either midday or midnight) exposed to natural light showed a clear day–night rhythm with increased nocturnal activity: significantly greater gape angle, increased exhalant pumping and had significantly higher valve adduction rates. However, circadian rhythms were less clear directly after anthropogenic food deprivation, in terms of the circadian rhythm in gape angle becoming significantly more apparent over the following days. Unlike mussels fed at midnight, those fed at midday displayed no significant change in gape angle from the hour before to the hour after they were fed, i.e. mussels given food at midday reacted to this food less than mussels fed at midnight. We suggest that independent of feeding time, laboratory mussels exposed to natural light and free from anthropogenic disturbance increase feeding activity at night because their circadian rhythms are strongly influenced by light levels. This study emphasises that the behaviour of animals in the laboratory and in the wild can be altered by anthropogenic disturbances such as vibrations caused by experimental setups and artificial illumination at night.     

The circadian system of Trypanosoma cruzi-infected mice

The effects of Chagas disease on the mammalian circadian system were studied in Trypanosoma cruzi-infected C57-Bl6J mice. Animals were inoculated with CAI or RA strains of T. cruzi or vehicle, parasitism confirmed by blood specimen visualization and locomotor activity rhythms analyzed by wheel-running recording. RA-strain infected mice exhibited significantly decreased amplitude of circadian rhythms, both under light-dark and constant dark conditions, probably due to motor deficiencies. CAI-treated animals showed normal locomotor activity rhythms. However, in these mice, reentrainment to a 6h phase shift of the LD cycle took significantly longer than controls, and application of 15min light pulses in DD produced smaller phase delays of the rhythms. All groups exhibited light-induced Fos expression in the suprachiasmatic nuclei. We conclude that the main effect of T. cruzi infection on the circadian system is an impairment of the motor output from the clock toward controlled rhythms, together with an effect on circadian visual sensitivity.     

Effects of starvation on the formation of daily growth increments in the otoliths of milkfish, Chanos chanos (Forsskål), larvae

The effects of starvation on daily growth and increment formation in the otolith were examined using a double oxytetracycline-labelling method on larval milkfish, Chanos chanos (Forsskål), reared under different feeding regimes. The results indicated that the differences in body and otolith growth between the larvae fed once and three times a day were not significant, and that the otolith growth increment was deposited daily in both groups of fed larvae. In contrast, the starved larvae grew at a slower rate than fed larvae in body length and otolith dimensions, and the otolith growth increment in the starved larvae was not deposited on a daily basis. After undergoing starvation, the larvae were unable to recover their normal growth either in otolith increment deposition or in body and otolith growth even though they were fed. Therefore, the application of ageing techniques based on counting otolith growth increments seems to be inaccurate for starved larvae.     

Effect of Pinealectomy on Re-entrainment of Air-gulping Activity in Indian Walking Catfish,Clarias batrachus

The present study was aimed to examine the effect of pinealectomy on re-entrainment of air-gulping activity in Indian walking catfish Clarias batrachus under various LD phases. Live Clarias batrachus of mixed sex (40-50g body weight) were procured from the local market and kept in the stock aquaria under the laboratory conditions for proper acclimation. Following 7 days of acclimation, animals were randomly selected from the stock aquaria and divided into 2 groups. The pineal was surgically removed from one group (P_xG) and the other group was treated as intact control group (IG). Both groups were exposed under LD 12:12 cycle inside the chronocubicles. In every 7 days LD schedule was delayed by 4 hours by lengthening of the light-on time. Both intact and pinealectomized animals are exhibited 24h rhythm in air-gulping behaviour and its entrainability irrespective of phase shifts of LD cycle. However, the phase angle of peak with reference to light on hour gradually decreased from phase 1 to phase 4, irrespective of treatment (intact/pinealectomized). Further, it has been noticed that the occurrence of multifrequency rhythm (24h, 12h and 6h) is being decreased from phase 1 to phase 4. It could be concluded that daily light-dark cycle plays an important role in modulating the rhythmic characteristics of air-breathing activity in Clarias batrachus and that pinealectomy does not modulate the effects of LD cycles on such activity.     

Circadian Phase Dependent Thermal Stimulation of Ovarian Recrudescence in Indian Catfish, Clarias batrachus

Female Clarias batrachus acclimated to long photoperiod (13L:11D), were subjected to 30º ± 1ºC thermopulses of either 6-hour or 12-hour duration at different phases of the LD cycle during the late resting phase (first week of January) of their annual reproductive cycle. Six-hour pulses were given either at 0600 or 1200 or 1800 or 0000. Twelve-hour thermopulses were given at 0600 or 1800. The long photoperiods were started at 0530 and that of ambient at 0630 coinciding with the average timing of sunrise that prevailed during the period of the study. The results indicate that exposure to long photoperiod or constant high temperature induced gonadal growth (GSI) and elevated testosterone and oestradiol levels in plasma. The high temperature was significantly more effective in its action. Further, combination of long photoperiod and high temperature produced the strongest gonadal stimulation as gauged from GSI and the levels of steroid hormones. Interestingly, 30ºC thermopulses of 12-h duration when given at 0600 to fish held under long photoperiod induced gonadal development of comparable magnitude as observed in response to constant high temperature under long photoperiod. Thermopulses (30ºC) of 6-h duration given at 0600 or 1200 also induced significant gonadal recrudescence but of much lesser magnitude. Thermopulses either of 6-h or 12-h duration at 1800 failed to elicit any change in the variables under study. The results of cosinor analysis performed on the responses to 6-h thermopulses also substantiate that there is a rhythm in the sensitivity of C. batrachus to thermopulses. Thus it appears that in this species temperature-induced gonadal recrudescence would occur only following coincidence of high temperature with the thermoinducible phase. The underlying mechanism of this phenomenon might be circadian in nature.     

Starvation and the Mating Success of Wild Male Mediterranean Fruit Flies (Diptera: Tephritidae)

Recent evidence suggests that the nutritional state of male Mediterranean fruit flies, Ceratitis capitata (Wied.) (medfly), is an important influence on various components of their reproductive biology, including mating success. The objective of the present study was to examine experimentally the effect of temporary starvation on the mating success of wild male C. capitata. Males were maintained on protein–sugar or sugar-only diets, and for each diet we compared the mating success of continuously fed males versus males starved for 18 or 24 h immediately before testing. In trials conducted on field-caged, host trees, males starved for 24 h obtained only about half as many matings as fed males for both diets. However, when the starvation period was 18 h, starved males reared on the protein–sugar diet mated significantly less frequently than fed males, whereas starved males reared on sugar mated as often as fed males. Measurements of male pheromone calling and female attraction revealed that reduced mating success likely reflected the decreased signaling activity of starved males.     

Effect of prior nutritional status on the activity of lipogenic enzymes in primary monolayer cultures of rat hepatocytes.

Effect of prior nutritional status of the animal on the activity of lipogenic enzymes and the fatty acid content of cultured hepatocytes was investigated. Hepatocytes were isolated from rats that were starved for 24 h ('starved') or continuously fed ('fed'), or starved for 48 h and then re-fed for 48 h ('re-fed') with a carbohydrate-rich fat-free diet, and maintained as monolayer cultures for 96 h in a serum-free glucose-rich medium (Waymouth's MB752/1) supplemented with insulin, dexamethasone and tri-iodothyronine. The fatty acid content and the activities of acetyl-CoA carboxylase, fatty acid synthase, glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase were determined initially at 3 h after plating and then every 24 h. Initially the activities of all the four enzymes were highest in hepatocytes isolated from the re-fed rats and lowest in those from the starved rats. With time in culture, the activity of all these enzymes increased severalfold (2-5, depending on the enzyme under consideration) in hepatocytes isolated from fed and starved rats, whereas there was a severalfold (2-5) decrease in the activity of these enzymes in hepatocytes isolated from re-fed rats. The initial fatty acid content of the hepatocytes from re-fed rats was 2-3 times that in the other two groups of hepatocytes. The fatty acid content seemed to increase in all three groups of hepatocytes during the 96 h in culture, but these apparent increases were not statistically significant.     

Some effects of early starvation on the survival and development of barnacle nauplii,Balanus improvisus (Darwin)

Newly hatched stage I–II nauplii of Balanus improvisus (Darwin) were “totally starved” (until death) or “partially starved” for the first 48 h and 96 h of their development. Daily mortality and molting were monitored throughout larval development in both starved and fed control groups. Fed control animals exhibited a largely synchronous molting pattern with instars of equal duration. Total starvation suppressed molting beyond stage II; 50% mortality occurred in ≈4 days at both 15 and 21 °C, while longest survival time was 7 days at 15 °C and 6 days at 21 °C. At 15 °C, partially starved nauplii retained the ability to complete naupliar development but at a slower overall rate and with increased mortality relative to controls. These effects were more pronounced in the 96-h group. Increased mortality of stage VI nauplii was evident in both partially starved groups (7.1% for 48 h, 18.8% for 96 h) relative to unstarved controls (3.1%). Stage II nauplii exhibited little resistance to starvation and survival potential may have decreased as soon as 24 h.     

Effects of thyroidectomy and starvation on the activity and properties of hepatic carnitine palmitoyltransferase.

1. Hepatic carnitine palmitoyltransferase activity was measured over a range of concentrations of palmitoyl-CoA and in the presence of several concentrations of the inhibitor malonyl-CoA. These measurements were made in mitochondria obtained from the livers of fed and starved (24 h) normal rats and of fed and starved thyroidectomized rats. 2. In the fed state thyroidectomy substantially decreased overt carnitine palmitoyltransferase activity and also decreased both the Hill coefficient and the s0.5 when palmitoyl-CoA concentration was varied as substrate. Thyroidectomy did not appreciably alter the inhibitory effect of malonyl-CoA on the enzyme. 3. Starvation increased overt carnitine palmitoyltransferase activity in both the fed and the thyroidectomized state. In percentage terms this response to starvation was substantially greater after thyroidectomy. In both the hypothyroid and normal states starvation decreased sensitivity to inhibition by malonyl-CoA.     

Effects of starvation on lung mechanics and biochemistry in young and old rats

Two groups of rats (young and old) were food-deprived for 3 wk and were compared with age-matched fed groups. Final body weight and dry and wet weights of lungs were significantly reduced in both young and old starved rats. As determined by saline volume-pressure (VP) curves, lungs of young starved rats accepted significantly less volume at all pressure levels compared with lungs of young fed rats. When expressed as a percent of maximum lung volume, the VP curve in young starved rats was significantly shifted upward at low lung volumes. In the old rats, the VP curves were similar in fed and starved rats. Total lung content of protein, DNA, crude connective tissue, hydroxyproline, and elastin were significantly reduced in young starved compared with young fed rats, whereas in old starved rats only protein and DNA contents were lower than those in old fed animals. It appears that in rapidly growing young rats starvation leads to growth retardation, loss of connective tissue components, and possibly reduction in tissue elastic forces at low lung volumes, whereas starvation has no significant effects on lung mechanics and connective tissue in old rats.