Nutritional characterization of Mucuna pruiriens: 2. In vitro ruminal fluid fermentability of Mucuna pruriens,Mucunal-dopa and soybean meal incubated with or without l-dopa

Three in vitro experiments were conducted to determine the rumen fermentability of Mucuna (M) pruriens (24 g 3,4-dihydroxy-l-phenylalanine (l-dopa)/kg dry matter (DM) and soybean meal treated with (SBD) or without (SB) 138 g l-dopa/kg DM). Additional objectives were to determine if l-dopa inhibits rumen fermentation, and if ruminal microbes can adapt to l-dopa or M. In Experiment 1, ground (1 mm) substrates were incubated in triplicate at 38 °C in 9 ml nutrient media and 1 ml rumen fluid in a series of six, 48 h, consecutive batch cultures. The first culture was inoculated with rumen fluid from two donor cows. Subsequent cultures were inoculated with fluid (1 ml) from the previous culture. The DM digestibility (DMD, 616 g/kg vs. 540 g/kg; P<0.01) and gas production (51.7 ml/g vs. 44.2 ml/g DM; P<0.05) were higher from fermentation of M versus SB but similar for SB and SBD (540 g/kg vs. 554 g/kg and 44.2 ml/g DM vs. 43.5 ml/g DM, respectively). The slopes of the relationships between DMD (g/kg) or gas production (ml/g DM) and fermentation period were not reduced by fermenting M (−0.014 DMD slope; 2.28 gas production slope) or SBD (−0.014 DMD slope; 0.459 gas production slope), instead of SB (−0.002 DMD slope; 1.039 gas production slope), indicating microbial adaptation to M and SBD. Total volatile fatty acid concentration (VFA; 53.7, 54.9 and 54.9 mmol/l) and molar proportions of VFA were similar among substrates. Gas production kinetics of M versus SB (Experiment 2), and SB versus SBD (Experiment 3) were also measured after substrates were incubated in triplicate in buffered rumen fluid for 24 h using a non-linear exponential model to fit the data. Residual l-dopa was measured after separate fermentation of substrates in triplicate for 0, 4, 8, 16 and 24 h. Fermentation of M versus SB produced more (P<0.05) gas (250 ml/g vs. 100 ml/g DM) and total VFA (203 mmol/l vs. 180 mmol/l) and a lower (P<0.05) acetate:propionate ratio (1.35 vs. 1.87; P<0.05). Adding l-dopa to SB increased (P<0.01) gas production (92 ml/g DM vs. 200 ml/g DM), and total VFA concentration (132 mmol/l vs. 188 mmol/l), but reduced (P<0.05) gas production rate (0.08 ml/h vs. 0.05 ml/h). The concentration of l-dopa in fermented M and SBD decreased by 53 and 47%, respectively during fermentation. In conclusion, M was more fermented than SB and degradation of l-dopa during ruminal fermentation and microbial adaptation to l-dopa were confirmed. Adding l-dopa to SB did not impair ruminal fermentation.     

Host‐seeking activity and avian host preferences of mosquitoes associated with West Nile virus transmission in the northeastern U.S.A.

Mosquito host‐seeking activity was studied using a custom‐designed trap to explore: (1) at which time interval of the night adult mosquito abatement would be most effective, and (2) if there exists an avian‐specific host‐seeking preference. Overnight trials using traps baited with dry ice showed that Aedes taeniorhynchus (Wiedemann) was most active at dusk and was then captured throughout the night. In contrast, Culex spp. (Cx. pipiens (Linnaeus) and Cx. restuans (Theobald) delayed most activity until about two h after dusk and were then captured through the night. This pattern suggests that management activities directed at adult Culex spp. would be most effective if initiated well after sunset. Mosquito capture rates in traps baited with birds in net bags were significantly greater than those with empty net bags, indicating that mosquitoes were attracted to the birds and not incidentally being sucked in by the custom trap's strong fan motor (Wilcoxon matched‐pairs signed‐ranks test, n= 24, t= 30, p < 0.05). Regression analysis showed that bird weight influenced mosquito attraction (r²= 0.21, p= 0.02). Trials with paired traps that contained different native bird species showed that Gray Catbirds, Dumatella carolinensis, attracted more mosquitoes than the heavier Northern Cardinals, Cardinalis cardinalis (paired samples t‐test, t= 2.58, df= 7, p= 0.04). However, attractiveness did not differ substantially among bird species, and Gray Catbirds did not attract more mosquitoes than all other birds combined as a group. American Robins, Turdus migratorius (n = 4) were comparable in attractiveness to other bird species, but not enough American Robins were captured for a comprehensive study of mosquito avian preference.     

Fate of transgenic DNA and evaluation of metabolic effects in goats fed genetically modified soybean and in their offsprings

The presence of DNA fragments in blood and milk from goats fed conventional (control) or Roundup Ready® soybean meal solvent extracted (s.e.; treated) was investigated by using a polymerase chain reaction approach. The same investigation was carried out on blood, skeletal muscle and organs from kids of both groups fed only dams’ milk until weaning. Moreover, the possible effects on cell metabolism were evaluated by determination of several specific enzymes in serum, heart, skeletal muscle, liver and kidney. Fragments of the multicopy chloroplast (trnL) gene were found in blood and milk samples from goats of both groups. In kids, the chloroplast fragments were found in samples of both groups. In samples, which proved positive for the presence of chloroplast DNA, fragments of the specific soybean single copy gene (lectin) were detected in several blood and milk samples. The same fragment was also found in control and treated groups of kids. Transgenic fragments were not found in those samples, which were found positive for chloroplast fragments of control groups of either goats or kids. On the contrary, in blood and milk of treated goats, fragments both of the 35S promoter and the CP4 epsps gene were detected. These fragments were also found in treated kids with a significant detection of the 35S promoter in liver, kidney and blood, and of the CP4 epsps gene fragment in liver, kidney, heart and muscle. A significant increase in lactic dehydrogenase, mainly concerning the lactic dehydrogenase-1 isoenzyme was found in heart, skeletal muscle and kidney of treated kids, thus suggesting a change in the local production of the enzyme. Finally, no significant differences were detected concerning kid body and organ weight.     

Insecticidal activity of spore-free mutants of Bacillus thuringiensis against the Indian meal moth and almond moth

Three oligosporogenic mutants of Bacillus thuringiensis were assayed for toxicity against larvae of the Indian meal moth, Plodia interpunctella, and the almond moth, Ephestia cautella. The results were compared with insecticidal activity obtained from the parent strain (HD-1) and two standard B. thuringiensis formulations (HD-1-S-1971 and HD-1-S-1980) against the same insect species. The toxicity of the sporeless mutant preparations was significantly diminished against the Indian meal moth (10- to 26-fold increase in LC₅₀) but exceeded the toxicity of the standards against the almond moth. The toxicities of the B. thuringiensis preparations toward the Indian meal moth were consistent with the number of spores in the test samples, but spores did not contribute to toxicity to E. cautella larvae. A rationale for basing dosage on soluble protein was demonstrated for use in situations where spores are not a contributing factor in toxicity.     

Volumetric feedback and the control of meal size in Schistocerca gregaria

The role and nature of volumetric feedbacks in the regulation of meal size was investigated for Schistocerca gregaria. Feedback from the anterior region of the crop was found to be important and denervation led to hyperphagia. Additional feedbacks were found from more posterior gut regions. Cannulating agar or paraffin into the mid- and hindgut caused a decrease in the size of a following meal. This effect was removed by cutting the ventral nerve cord anterior to the terminal abdominal ganglion. Increasing the blood volume by injections of isotonic saline into the haemocoel was also found to decrease subsequent meal size. This effect was not mediated via the gut or body wall stretch receptors and possible mechanisms are discussed. The results for S. gregaria are compared with those obtained previously from Locusta migratoria.

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Résumé L'étude de la nature et du rôle des feed backs volumétriques dans la régulation de la taille du repas a été effectuée chez S. gregaria. Un feed back, originaire de la région antérieure du jabot, s'est révélé important et la dénervation a provoqué l'hyperphagie. Des feed backs supplémentaires ont été décelés dans des régions plus postérieures du jabot. De l'agar ou de la paraffine canulés dans la jabot moyen et postérieur ont provoqué une diminution de la taille du repas suivant. Ces effets sont supprimés avec la section de la corde nerveuse centrale avant le ganglion abdominal terminal. L'augmentation du volume sanguin par des injections de solution saline isotonique a aussi diminué la taille du repas suivant. Cet effet n'est pas transmis via des récepteurs de tension du jabot ou de la paroi du corps; des mécanismes envisageables sont discutés. Ces résultats avec S. gregaria sont comparés à ceux obtenus antérieurement avec Locusta migratoria.

     

Cellular localisation of aminopeptidase in the midgut of Rhodnius prolixus Stål (Hemiptera:Reduviidae) during blood digestion

Summary By use of the artificial substrate leucyl--naphthylamide, aminopeptidase was localised in the midgut cells of the haematophagous insect Rhodnius prolixus before and at various times up to 25 days after a meal of rabbit blood. The enzyme was primarily associated with the membranes of the microvilli, with extracellular membrane layers and with the lysosomes of the midgut cells. Aminopeptidase activity was also detected on the rough endoplasmic reticulum and at the periphery of intracellular storage vesicles. The absence of aminopeptidase on the microvilli of the crop supports the conclusion that the crop is not involved in the digestion of blood-meal proteins and that protein digestion is restricted to the intestine. The sites of localisation are in accordance with models for the spatial separation of digestive enzymes in the midgut of several non-haematophagous insects, and this suggests that aminopeptidase plays a major role in the terminal digestion of the blood meal. The changes in enzyme localisation during the digestive period correlate with previously described cycles of digestive-enzyme activity and changes in midgut ultrastructure. A model for blood protein digestion in R. prolixus is described.     

Chronobiological Aspects of Weight Loss in Obesity: Effects of Different Meal Timing Regimens

A series of short- and long-lasting experimental protocols of different meal timing regimes were performed in obese subjects to assess the possible occurrence of (1) a different metabolic fate of nutrients; (2) a phase shift of circadian rhythms of metabolic and hormonal parameters strictly related to nutrition; (3) a different weight loss. (A) In a short-lasting protocol (3 days) IS obese subjects were fed a hypocaloric diet (684 kcal/day) (a) at 10 hr only, (b) at 1800 hr only; (c) at 1000 hr, 1400 hr and 1800 hr, or (d) studied during a 36-hr fasting. Measures of calorimetry (R.Q., CHO and lipid oxidations, energy expenditure), hormones (plasma Cortisol, insulin, HGH, urinary catecholamines), urinary electrolytes (Na, K) and vital parameters (body temperature, heart rate, blood pressure) were carried out at 4-hr intervals for three days. A significantly higher lipid oxidation and a lower CHO oxidation were documented with the meal at 1800 hr, in comparison with the meal at 1000 hr. CHO and lipid oxidation circadian rhythms appeared the most affected by meal timing. (B) In a long-lasting protocol (18 days) 10 obese subjects were fed the same hypocaloric diet (a) at 1000 hr only and (b) at 1800 hr only. Calorimetric measures were performed every other day for 2 hr preceding each meal. Before and after the 18-days single meal period, body temperature, plasma Cortisol, PRL and TSH were recorded (δt = 4 hr). A higher lipid oxidation and a lower CHO oxidation were again demonstrated with the meal at 18 hr. Minimal changes of hormonal circadian rhythms were documented suggesting that the hypothalamus-hypophysis network is scarcely affected by meal timing. Weight loss did not vary in both short- and long-term protocol.     

Meal Scheduling Modulation of Circadian Rhythm of Phototactic Behaviour in Cave Dwelling Fish

Phototactic responses of three groups, each of 16 cave fishes, were observed at 4-hr intervals beginning at 0900, following S days of acclimation inside specially designed aquaria. The latter consisted of two parts, a photic zone and an aphotic zone. The first group of fishes were fed ad libitum, while the second and third group of fishes were fed with minced mutton and/or liver delivered in the photic zone between 0700-1100, and in the aphotic zone between 1900-2100. The percentage of fishes present in the photic zone at a given time was used as a measure of their phototactic responses. Single cosinor analysis of these data has revealed, for the first time, a circadian rhythmicity in the phototactic behaviour of a cavernicolous organism. However, imposition of restricted feeding schedules shifted the peak time of the circadian rhythm in phototactic responses. Meal scheduling may thus be an effective synchronizer for this behavioural rhythm.     

Production of phytate-hydrolysing enzyme by some fungi

Eighty-four fungi from twenty five species have been examined for the production of extracellular enzymes capable of hydrolysing phytate (3-phytase, myo-inositol hexakisphosphate 3-phosphohydrolase, EC 3.1.3.8, and 6-phytase, myo-inositol hexakisphosphate 6-phosphohydrolase, EC 3.1.3.26) when grown in: (1) rapeseed meal (RSM); (2) a semisynthetic medium containing phytate as the sole phosphorus source (PSM); (3) potato dextrose broth (PDB). Although 58 active strains showed substantial activity, results in either of the media were of no value in indicating activity in RSM. There was no relationship between the ability of a fungus to hydrolyse phytate and its taxonomic position. Aspergillus ficuum NRRL 3135 had the greatest activity in the synthetic medium, and was relatively active in RSM. The extracellular enzyme had maximum activity after 10 days growth in PSM and had a temperature optimum of 55°C. Two pH optima were noted at pH 2.0 and 5.5. Inorganic phosphate inhibited enzyme production; ammonia ions were a better nitrogen source than nitrate or urea.