Impact of a shift work-like lighting schedule on the functioning of the circadian system in the short-lived fish Nothobranchius furzeri.

Adult Nothobranchius furzeri of the MZM-04/10 strain were individually kept and subjected to a "5 + 2" shifting lighting schedule (SHIFT) for 8 weeks in order to evaluate the desynchronizing effects of a simulated human-like shift-work schedule on the functioning of the circadian system (CS). With this aim, sixteen 21-week-old N. furzeri were placed into a Morning, Night and Evening schedule (lights on from 08:00 to 16:00, 00:00 to 08:00 and 16:00 to 00:00 h, respectively) and fed once a day in the middle of the corresponding photophase (12:00, 04:00 and 20:00 h, respectively). Then, in the weekends (2 days), fish were always returned to the Morning shift. As controls, 16 fish were maintained under a non-shifting LD cycle condition (CONTROL) throughout the whole experiment, with lights on from 08:00 to 16:00 h. Rest-activity rhythm (RAR) of fish subjected to SHIFT showed several symptoms of chronodisruption, such as a decrease in the percentage of diurnal activity and a reduction of the relative amplitude and the circadian function index with time. When a periodogram analysis was performed, RAR of N. furzeri under SHIFT conditions showed up to three separate circadian components: one longer than 24 h (26.5 h) that followed the weekly 8 h delays; a short-period component (~23 h) that was related to the weekend's phase advances, and finally, a 24 h component. The shifting LD schedule also affected fish CS at a molecular level, with several significant differences in the expression of core genes of the molecular clock (bmal1, clock, rorα, rev-erbα) between SHIFT and CONTROL animals. RAR impairment along with changes in clock gene expression could be associated with high stress and accelerated aging in these fish.

Age-related changes in fatty acid profile and locomotor activity rhythms in Nothobranchius korthausae.

The life cycle of Nothobranchius korthausae, a Cyprinodontiformes fish, was studied in our laboratory to characterise the ageing process. Some morphological changes, such as spine curvature, skin colour, and fin and eye appearance are described. Growth and survival curves reflected a fast life cycle with rapid initial growth until 4weeks of age, after which the fish grew more slowly before reaching their final size in week 40. Senescence onset was established at week 48 with a decrease in spawn size and viability and a general decline in the animal's appearance (weight and colouration losses, caudal fin degradation, and cataractogenesis). The fatty acid composition changed with age, with high unsaturation in the adult stage as reflected by a high peroxidation index, a condition that is associated with high susceptibility to oxidative damage if elevated reactive oxygen species (ROS) production occurs. Senescent fish had an increase in monounsaturated fatty acid proportions and a lower peroxidation index (226.5±19.7 in adults versus 120.2±19.1 in senescent fish, P<0.05). The circadian system, as reflected by locomotor activity rhythms, showed noticeable changes with age. Twenty-four-week-old fish (adults) had a robust diurnal rhythm that showed a decrease in total activity, an increase in rhythm fragmentation, and a fall in amplitude and regularity with age. Changes were clearly reflected in the Circadian Function Index variations (0.56, 0.47 and 0.25 at 24, 48 and 72weeks of age, respectively). In conclusion, N. korthausae appears to be a species with appropriate characteristics for ageing studies because it manifests clear signs of progressive ageing. Comparing species of Nothobranchius genus with different lifespans may be useful for increasing our understanding of the ageing process.

Energy intake and macronutrient selection in sharpsnout seabream (Diplodus puntazzo) challenged with fat dilution and fat deprivation using encapsulated diets.

Sharpsnout seabream fed pure macronutrient capsules were challenged to fat dilution and fat deprivation in order to investigate the effects of fat level on energy intake regulation and macronutrient selection by fish, as they lack oropharyngeal chemosensory information from the diet. During the control phase, the fish were fed three individually encapsulated macronutrients, from which they composed a diet containing 67.36% protein (P), 19.08% carbohydrates (CH) and 13.57% fat (F), in terms of macronutrient weight intake percentage. During the second phase of the experiment, a lipid content reduction in F capsules from 55.0% to 13.4% did not significantly modify this selection pattern, energy ingestion or the number of capsules ingested of each macronutrient. During the third phase, in which they were subjected to fat deprivation, starting on almost the first day, the fish increased their total energy intake and total ingested number of capsules. These results reveal that fish are capable of distinguishing and selecting each of the three macronutrients contained in gelatine capsules, and that fish selection of a balanced diet from pure macronutrients is remarkably stable. Fish are capable of sustaining their macronutrient selection pattern and energy intake with very low amounts of fat in their diets (Phase 2). A certain instability in the initial P, CH and energy intake was only observed when fat was totally deprived (Phase 3), which resulted in higher values than those observed in Phase 1. In order to examine any possible effects of diet encapsulation, digestibility assays were performed in a second experiment. The fish were divided into two experimental groups and fed the same complete commercial diet, the only difference being the way it was presented to each group (pelleted or encapsulated). No statistical differences between the experimental groups were found with regards to both apparent digestibility coefficients and fish growth.

Macronutrient selection through post-ingestive signals in sharpsnout seabream fed gelatine capsules and challenged with protein dilution.

Sharpsnout seabream ability for macronutrient self-selection was studied using gelatine capsules containing pure macronutrients. In particular, the existence of non-oropharyngeal chemosensory pathways involved on protein (P), fat (F) and carbohydrate (CH) selection, as well as sharpsnout seabream response to dietary protein dilution were investigated. In a sequence of experimental phases, sharpsnout seabream were fed a pelleted complete diet, an encapsulated complete diet or a combination of separately encapsulated pure macronutrients. In order to induce associative learning, capsules containing a given macronutrient were paired with a particular colour. The animals composed a diet containing 62.7% P, 21.3% CH and 16.0% F, in terms of macronutrient percentage intake, and this selection pattern was maintained throughout all experimental phases. In a second experiment, individually kept sharpsnout seabream were challenged with protein dilution. After protein capsules were diluted (from 91.9% P to 56.3% P) with cellulose, the animals increased their protein intake to compensate for dilution in such a way that their energy intake was not significantly modified (17.4 kJ/100 g BW vs. 17.6 kJ/100 g BW after dilution). These results show that sharpsnout seabream feeding on encapsulated diets are able to select and maintain a particular diet composition, as well as sustain their energy intake, without using the diet's oropharyngeal chemosensory properties. Moreover, they were also able to maintain their protein intake after dilution, which highlights the importance of this macronutrient in this omnivorous species.

Effects of intense exercise training on rainbow trout growth, body composition and metabolic responses.

Cultured fish are bound to experience a variety of potentially stressful situations. In principle, stress may be reduced by adapting fish to intense exercise. This article addresses the effects of chase-induced intense exercise training on rainbow trout. Training reduced fish growth and worsened their nutritional use of food, but had no effect on food intake, biometry or body composition, indicating that a significant fraction of the diet's energy was being used for exercising rather than growth. Regarding metabolic responses, training significantly counterbalanced lactate, glucose and cortisol plasma concentration increases induced by intense exercise, while completely abolishing that of plasmatic proteins. These data suggest that this type of training adversely affects fish growth and food conversion, whereas it improves their metabolic response capacity and reduces the stress component of forced exercise.

Influence of exercise on plasma and muscle free amino acids in trained rainbow trout.

Variations in the concentration of free amino acids in the muscle and plasma of trout submitted to 5 minutes of intense exercise have been studied. The responses of untrained fish and those trained performing the same type of exercise twice daily for 28 days are compared. Total amino acid concentrations in muscle tend to diminish after intense exercise. Significant decreases are observed in muscle content of alanine, beta-alanine, isoleucine and ornithine. Plasma amino acids tend to increase after exercise with significant differences in glutamate, GABA, methionine and NH4+. The small variations due to intense exercise suggest that the amino acids are mobilised. Training led to a decrease in total amino acid concentration in plasma but not in muscle, where levels of aspartate and ornithine increased. This suggests a metabolic adaptation to exercise, with amino acid level retention in the muscle.

Trout plasma metabolites, blood factors and spleen contraction: responses to exercise.

Fish react to handling and capture with a burst of exercise that affects them deeply. The present study examines the effect of such severe exercise and the time course of recovery on the hematology (including spleen response) and metabolism of a population of cultured rainbow trout. Exercise was induced by continuous chasing for 5 min when the trout showed signs of exhaustion. Such exercise led to spleen contraction and an increase in haematocrit values. Carbohydrates were mobilized and anaerobic glycolysis produced lactate without significant effect on lipid metabolism. The conclusion is reached that the respiratory properties of rainbow trout blood do not change following severe exercise, while muscle anaerobic metabolism is slightly activated as deduced from the fast and short lactacidemia observed, which may have been related to a reduced stressing component, as the exercise was performed in the same environment in which the fish were reared.

Sexual abuse within the marital relationship.

The authors aim to characterise sexual abuse within the marital relationship. They analyzed the reports of the clinical examinations performed at the Institute of Legal Medicine of Oporto on victims of sexual abuse from January 1990 until December 1994. They also randomly interviewed victims of physical abuse at the Institute to determine if sexual abuse was performed by their partners. This study shows that complaints are rarely reported by victims of sexual abuse in marital relationships, and in all the cases studied where sexual abuse was verified it was accompanied by other forms of abuse.

Factors simultaneously affecting the growth of the rainbow trout (Oncorhynchus mykiss).

Growth of rainbow trout (Oncorhynchus mykiss) is examined in relation to simultaneous variation of initial weight, initial density, temperature and time (days). Twenty-six lots of medium weight were set up, ranging from 92 to 372 g; initial density ranged from 7.2 to 38.8 kg/m3; and temperatures from 14 degrees C in winter to 20 degrees well into spring. Experimental periods carried out were of 7, 20, 38 and 26 days duration. By multiple correlation and regression analysis, predictive equations for increments in weight, final density were obtained in relation to the afore mentioned variables and ranges. The values of multiple correlation obtained were highly significant. We confirmed that the effect of each factor on growth is modified by the simultaneous action of the other variables.

Simultaneous effect of initial weight, initial crowding, temperature and O2 concentration on the nutritional use of food by rainbow trout (Oncorhynchus mykiss).

The simultaneous effects of initial weight, initial crowding, temperature and O2 concentration on the following ratios: relative growth rate percent (RGRP), feed efficiency (FE), protein efficiency ratio (PER) and protein productive value (PPV) were studied in the rainbow trout. Multivariant equations were obtained for each of the mentioned indices. The joint effects of these factors were evidenced by means of a multiple correlation analysis. The influence of temperature and, to a lesser extent, of crowding, and O2 concentration on the nutritional use of food by the trout was demonstrated, their fundamental dependence on factors extrinsic to the animal being underlined. The non proportional changes in PER and PPV as temperature rises revealed that an increasing part of the ingested aminoacids were used for synthesis of fat, non for proteins edification.

Effect of simultaneous variation of weight, density, temperature and O2 concentration on rainbow trout (Oncorhynchus mykiss) body composition.

The simultaneous effect of weight, initial density (kg/m3, temperature and O2 concentration on rainbow trout body composition (fat, protein, moisture and ash) has been studied. In 3 successive experimental phases fish were kept in different lots of varying initial weight (178-372 g), initial density (7.2-38.8 kg/m3) and temperature (15-20 degrees C). Simple correlations were statistically significant for weight vs fat (r = 0.56; P less than 0.001) and moisture (r = -0.57; P less than 0.001); temperature vs fat (r = 0.73; P less than 0.001) moisture (r = -0.73; P less than 0.001) and ash (r = -0.26; P less than 0.02); and O2 concentration vs fat (r = 0.22; P less than 0.05). Multivariant equations for the different compounds were obtained. Only fat and moisture percentages showed significant differences (rm = 0.75; P less than 0.00005); an inverse relation existing between them (r = -0.94; P less than 0.001). Temperature is the factor which has the strongest influence on fat and moisture when it varies simultaneously with weight, initial density and O2 concentration, which is shown by its equation coefficients (P less than 0.00005).

Histoplasmosis in individuals with acquired immunodeficiency syndrome (AIDS): report of six cases with cutaneous-mucosal involvement.

The authors report the first six cases of disseminated histoplasmosis and acquired immunodeficiency syndrome (AIDS) seen at the University Hospital, Faculty of Medicine of Ribeirão Preto, University of São Paulo, from 1987 to 1989, with emphasis on dermatological clinical manifestations, nasal mucosa lesions and treatment. The mycosis was the first manifestation of AIDS in four patients. It is concluded that biopsies of the lesions for histopathologic study and fungal culture are important for diagnosis.

Histochemical determination of muscle fiber types in locomotor muscles of anuran amphibians.

1. A histochemical study using myosin ATPase, succinate dehydrogenase and alpha-glycerophosphate dehydrogenase reactions and a morphometric analysis with image analyser, was carried out in sartorius and gastrocnemius muscles of two anuran species, Rana perezi and Bufo calamita, that show different locomotor activities. 2. Four types of muscle fiber were found. There were interspecific variations in their proportions, with a predominance of oxidative muscle fibers in Bufo calamita. 3. These results agree with those obtained previously for the metabolic profile of several tissues from both species and point to a clear metabolic basis for the differences in locomotor activities between these two species.

Seasonal changes in the activities of glycolytic enzymes from liver and skeletal muscle of Rana perezi.

Glycogen phosphorylase (GP), Hexokinase (HK), Phosphofructokinase (PFK), Pyruvate kinase (PK) and Lactate dehydrogenase (LDH) activities from skeletal muscle and liver were measured in Rana perezi for the four seasons of the year. Skeletal muscle showed a decrease in PFK, PK and LDH activity during winter and summer. Liver displayed an increase in GP activity in spring and in PK and LDH in autumn.