Effect of Evening Exposure to Dim or Bright Light on the Digestion of Carbohydrate in the Supper Meal

In a previous study we found that daytime exposure to bright as compared to dim light exerted a beneficial effect on the digestion of the evening meal. This finding prompted us to examine whether the digestion of the evening meal is also affected by evening light intensity. Subjects lived in light of 200 lux during the daytime (08:00–17:00 h) and took their evening meal at 17:00 h under 20 lux (evening dim‐light condition: 17:00–02:00 h) or 2000 lux (evening bright‐light condition: 17:00–02:00 h) until retiring at 02:00 h. Assessment of carbohydrate digestion of the evening meal was accomplished by a breath hydrogen test that is indicative of the malabsorption of dietary carbohydrate. Hydrogen excretion in the breath in the evening under the dim‐light condition was significantly less than under the bright‐light condition (p < 0.05). This finding is the opposite to that obtained in previous experiments in which subjects were exposed to the different intensities of light during the daytime, and indicates that the exposure to dim light in the evening exerts a better effect on carbohydrate digestion in the evening meal than does the exposure to bright light.     

Preliminary investigation of the digestive enzymes in Pagellus erythrinus (Linneo 1758) larvae

To gain a better understanding of the role of the olfactory system in sexual selection, we conducted a series of behavioral assays to correlate the number of aesthetascs with pre-copulatory behavior and male mating success in a protandrous simultaneous hermaphroditic shrimp, Lysmata wurdemanni. We demonstrated that male pre-copulatory behavior and mating success were closely associated with the number of aesthetascs. Individuals with a higher numbers of aesthetascs started approaching/searching females earlier and had higher mating success than ones with lower numbers. Sexual selection theory predicts that small males in a pure searching mating system have advantages to mate with females because they are cryptic and agile. However, our results show that aesthetasc number, rather than shrimp size, may be the major factor in determining male mating success.     

Carnivorous pitcher plants: insights in an old topic

Plant insect interactions are usually recognized as a scenario where herbivorous insects feed on a host plant. However, also the opposite situation is known, where plants feed on insects. Carnivorous pitcher plants of the genus Nepenthes as well as other pitcher plants obtain many nutrients from caught insect prey. Special features of the pitcher traps’ surface are responsible for attraction and trapping insects. Once caught, the prey is digested in the fluid of the pitchers to release nutrients and make them available for the plant. Nutrients are taken up by special glands localized on the inner surface of the pitchers. These glands also secrete the hydrolyzing enzymes into the digestion fluid. Although this is known for more than 100 years, our knowledge of the pitcher fluid composition is still limited. Only in recent years some enzymes have been purified from the pitcher fluid and their corresponding genes could be identified. Among them, many pathogenesis-related proteins have been identified, most of which exhibiting hydrolytic activities. The role of these proteins as well as the role of secondary metabolites, which have been identified in the pitcher fluid, is discussed in general and in the context of further studies on carnivorous plants that might give answers to basic questions in plant biology.     

Impact of feed restriction and fragmented feed distribution on performance,intake behaviour and digestion of the growing rabbit

Postweaning feed restriction preserves rabbit digestive health after weaning, but the underlying physiological mechanisms are not yet understood. To elucidate whether the feeding intake pattern modification related to feed restriction might be involved, we studied the effects of both feed intake quantity and intake frequency. Animals were allotted at weaning (28 d old) in a 2 × 2 factorial design: feed intake quantity (AL = ad libitum vs R = 75% of AL) and fragmented feed distribution (FFD) (1 vs 13 distributions), thus forming four groups (AL1, AL13, R1 and R13). New Zealand White growing rabbits were used from weaning to slaughter (70 d old), to analyse mortality, morbidity, performance, intake behaviour, digestion and microbial activity. Seven days after starting feed restriction (35 d old, group R1), rabbits consumed 44% of the feed within 2 h, 65% in 4 h and in 7 h over 95%. Over the 28–70 d period, mortality was low (5.3%) while morbidity averaged 18.5% and neither was affected by treatment. However, FFD tended to decrease the morbidity rate during the first 14 days after weaning (P = 0.06). Feed conversion (28–70 d) was improved by restriction (+15%, P < 0.001) and by FFD (+5%, P < 0.001). Nutrient digestibility was improved by restriction (+10% for energy, P < 0.01), but not by FFD. Fragmented feed distribution led to a lower stomachal pH, in the antrum (1.48 vs 2.13, P < 0.001) and in the fundus (1.52 vs 2.63, P < 0.001), while a higher pH was found in the caecum (6.07 vs 5.86, P < 0.001). Butyrate proportion in the caecum was reduced by four units for restricted groups. Fragmented feed distribution reduced the caecal VFA concentration by 23% within restricted rabbit groups only. A similar interaction between intake level and FFD was observed for fibrolytic activity (cellulase and xylanase). The diversity of caecal bacterial community was not modified by either of the two factors studied. Globally, fragmented meals have no major impacts on the caecal microbial activity, diversity, and thus would not be implicated in the better resistance of restricted rabbit to digestive troubles.     

The effects of temperature on metabolic interaction between digestion and locomotion in juveniles of three cyprinid fish (Carassius auratus, Cyprinus carpio and Spinibarbus sinensis)

To test whether the effects of temperature on the metabolic mode changed among different fish species, we investigated the specific dynamic action (SDA) and swimming performance of fasting and fed fish at 15 and 25°C in three juvenile Cyprinidae fish species: goldfish (Carassius auratus), common carp (Cyprinus carpio) and qingbo (Spinibarbus sinensis). Both taxon and temperature had significant effects on the resting oxygen consumption rate (M˙O(rest)), SDA and swimming performance (p<0.05). In addition, the effect of temperature differed significantly among the different species (interaction effect, p<0.05). Under the low temperature condition, digestion had no effect on either critical swimming speed (U(crit)) or the active MO(2) (MO(active)) for all fish species (additive metabolic mode). When the temperature was increased from 15 to 25°C, the metabolic scope (MS) for digestion increased approximately 182, 49 and 17%, and the MS for locomotion increased approximately 129, 58 and 138% in goldfish, common carp and qingbo, respectively. The total metabolic demands for both digestion and locomotion (i.e., the sum of digestive MS and locomotive MS) increased approximately 143, 56 and 112% in goldfish, common carp and qingbo, respectively. The total MS for both digestion and locomotion (the difference between MO(active) in fed fish and MO(rest) in fasting fish) increased approximately 106, 58 and 78% in goldfish, common carp and qingbo, respectively. Thus, the MS for locomotion in fed goldfish decreased due to the large increase in digestive function at the high temperature, and the U(crit) of fed goldfish decreased by 11% compared to that of fasting fish (p<0.05) (digestion-priory metabolic mode). The metabolic mode of qingbo changed to locomotion-priority mode, as illustrated by the large increase in locomotive MS in response to the increase in temperature. In the common carp, temperature had no effect on metabolic mode as illustrated by the parallel increases in cardio-respiratory capacity and metabolic capacity of digestive and locomotive organs. A discussion on the changes in metabolic mode in response to temperature and its possible relationship with the metabolic characteristics of a given fish species was also documented in this paper.     

Cholecystokinin (CCK) in Atlantic herring (Clupea harengus L.) - ontogeny and effects of feeding and diurnal rhythms

Neural and alimentary cholecystokinin (CCK) levels in Atlantic herring, Clupea harengus, were analyzed from hatching to 40days after hatching (DAH). The head compartment representing the neural pool was quantitatively dominant (>80% of the total CCK content) while the digestive tract pool represented 6-10%. During ontogeny the CCK level in whole larvae increased almost 15-fold from 0 to 40 DAH, being particularly marked from 14 to 20 DAH. Larvae of 24 to 26 DAH were examined for potential occurrence of a circadian rhythm and to analyze the effects of feeding. Fed and fasted larvae were significantly different, where fed larvae showed higher CCK levels. There were large fluctuations in CCK levels analyzed at 3h intervals without an apparent diurnal pattern. Shorter sampling intervals of 1h in the morning when lights were switched gradually on and food was offered to the larvae demonstrated a marked drop in the relative gut CCK levels and a concurrent increase in the CCK carcass to gut ratio, 1h after introduction of food followed by a return to prefeeding levels after 2h. This response probably results from a release and re-synthesis of CCK in the gut after initiation of feeding. Taken together, these results support earlier reports that CCK participates in the regulation of digestive processes in herring larvae, but CCK does not seem to have a circadian rhythm independent of feeding.     

Optimization of cell lysis and protein digestion protocols for the analysis of HeLa S3 cells by LC-MS/MS

In order to maximize the number of proteins identified from Hela S3 cell lysate we tested various cell lysis, protein precipitation and digestion protocols. First, we compared three different lysis buffers, two mechanical cell disruption methods and two precipitation methods. Then, we tested six different in-solution digestion protocols, three different in-gel digestion protocols and ten different peptide extraction protocols. The result is a proposal for an optimized protocol to prepare the whole cell lysate samples from HeLa S3 cells.     

Retention of different-sized particles and derived gut fill estimate in tammar wallabies (Macropus eugenii): physiological and methodological considerations

The capacity of the digestive tract is an important parameter in understanding digestive adaptations, particularly in herbivores. Measures of this capacity (‘gut fill’) are commonly performed in killed animals, which has ethical and logistical implications. Alternatively, dry matter gut contents (DMC) can be estimated in live animals from food intake, digesta retention and digestibility, based on physical principles (Holleman and White, Can. J. Zool. 67, 488-494, 1989). Although this method has been used to some extent, it still awaits thorough validation. Here we estimated DMC in seven tammar wallabies during 5-day feeding trials and compared the results to those gained from dissections immediately after the trials. Calculated DMC exceeded that actually measured by 29 ± 22%. A closer inspection of the data suggested that this was partly due to the fact that DMC as measured by dissection is susceptible to short-term influences such as daily variation in food intake, whereas the calculated DMC represents an integrative measure over the whole period of the feeding trial. Correlations between both the measured digesta retention times, and the calculated DMC, with the measured wet contents mass suggest that it is particularly the DMC determined via dissection that needs to be measured with care. For a comparison of gut capacities, the calculated DMC therefore can be considered adequate, but should for a more widespread use be validated in further studies including more species and experimental regimes controlling food intake variation. Additionally, we tested whether very small (100-500 μm) and small (500-1000 μm) particles were retained differently in the tammar wallabies. There was no indication of such a difference. Whether the macropod forestomach selectively passes a certain particle fraction (that represents microbes) with the generally faster-passing fluids remains to be investigated with even smaller markers, e.g. labelled bacteria.     

Sodium-deoxycholate-assisted tryptic digestion and identification of proteolytically resistant proteins

Identification of proteolytically resistant proteins with compact molecular structure and/or poor water solubility is a challenge in current proteomic study. In this study, sodium deoxycholate (SDC)-assisted tryptic digestion and identification of proteolytically resistant myoglobin and integral membrane proteins were systematically investigated. When the effect of SDC up to 10% on trypsin activity was investigated, little decrease in the trypsin activity was observed in 1% SDC solution, 2-5% SDC decreased the enzyme activity only by about 13.6%, and even in the presence of 10% SDC trypsin still retained 77.4% of its activity. Matrix-assisted laser desorption ionization time of flight mass spectrometry analysis showed that SDC could be removed from sample solution with acid treatment followed by centrifugation, and the remaining SDC, if any, had little effect on mass spectrometry analysis with regard to the number and signal/noise ratio of ions in the mass spectra. Compared with urea and methanol, two other commonly used additives in addition to SDS in proteomic analysis, SDC improved more efficiently the denaturation, solubilization, and tryptic digestion of proteins, particularly proteolytically resistant myoglobin and integral membrane proteins, thereby enhancing the efficiency of their identification with regard to the number of identified proteins and unique peptides and the sequence coverage of matched proteins.