Effects of nitrate and phosphate availabilities on growth, photosynthesis and pigment and protein contents in colour strains of Hypnea musciformis (Wulfen in Jacqu.) J.V. Lamour. (Gigartinales, Rhodophyta)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Net and metabolizable protein requirements for body weight gain in hair and wool lambs

Protein is one of the limiting factors in animal production, and the knowledge of protein requirements by livestock is crucial for the success of a commercial animal raising enterprise. Thirty-four castrated lambs, 17 of them F1 Ideal x lie de France wool lambs and the remaining ones were Santa Ines hair lambs, with homogeneous initial BW, were used in the experiment. Five animals from each genotype were slaughtered in the beginning of the experimental period and used as reference. Diets (D) were composed of concentrate mix (C) and Cynodon sp. c.v. Tifton 85 hay (R), combined in three different ratios: D1 =60CAOR; D2=40C:60R and D3=20C:80R. Animals of each group of three lambs, that showed simultaneously an initial BW of 20 +/- 0.14 kg at the beginning of the dietary regimen, were slaughtered when one of them reached 35 kg, what always happened to be the one fed with D1. Net requirements for BW gain in wool lamb, fleece-free, ranged from 101 to 110 g of protein/kg BW, and for hair lamb ranged from 1 10 to 118 g of protein/kg BW. Net protein requirements for wool production ranged from 634 to 642 g/kg of produced wool. Hair lambs presented a 7.8-9.5% higher estimated net protein requirements than wool lambs, according to BW and daily weight gain (DG). Total net protein requirements for Santa Ines and wool lambs, with 30 kg of initial B W and an approximate 200 g mean DG, were 48.5 and 45.4 g/day, respectively. Metabolizable protein requirements for Santa Ines and wool lambs, with 20 kg of initial BW and an approximate 200 g mean DG were 59.4 g and 76.5 -/day, respectively. Net protein requirements for wool production was 64g/100g of produced wool. Thus, under the conditions of this experiment, it is concluded that hair lambs showed a higher concentration of protein in the body, more efficient use of the ingested protein and a consequent additional BW gain when fed isoproteic diets as compared to F1 Ideal x Ile de France wool lambs. (c) 2006 Elsevier B.V All rights reserved.

Net requirements of protein and energy for maintenance of wool and hair lambs in a tropical region

The objective of this work was to evaluate the total endogenous N losses, and protein and energy net requirements for maintenance in growing lambs. Thirty-four castrated lambs, 17 F1 Ideal X Ile de France wool and 17 Santa Inas hair lambs, averaging 20 +/- 0.14 kg BW, were used in the experiment. Five animals from each genotype were slaughtered at the beginning of the experiment and taken as controls. Diets (D) were composed of concentrate mix (C) and Cynodon sp. c.v. Tifton 85 hay (R), combined in three different ratios: D1 = 60C:40R; D2 = 40C:60R and D3 = 20C:80R. Animals of each group of three lambs, that showed BW of 20 kg at the beginning of the dietary regimen, were slaughtered when one of them reached 35 kg, what always happened to be the one fed with D1. Total endogenous N losses estimated for wool lambs were 250 mg N/kg BW0.75. For hair lambs, total endogenous N losses reached 324 mg N/kg BW0.75 . Hair lambs showed higher (P < 0.01) (29.9%) net requirements of protein for maintenance than wool lambs. In contrast, net energy (NE) requirement for maintenance was similar (P > 0.05) for both genotypes (74.27 kcal/kg BW0.75 per day), the average of the antilog of the two intercept values obtained from the estimated regression equations of heat production for zero metabolizable energy (ME) consumption. Further studies should be done to check if this trend is also true for metabolizable energy and protein in animals exhibiting BW gains in tropical region. (C) 2003 Elsevier B.V. B.V. All rights reserved.

Increased production of beta-1,3 glucanase and proteins in Bipolaris sorokiniana pathosystem treated using commercial xanthan gum

Barley plants (cultivars Embrapa 127, 128 and 129) treated with xanthan gum, and with different time intervals between the administration of the inducer and the pathogen, demonstrated induction of resistance against Bipolaris sorokiniana. Induction was shown to have local and systemic action. In order to prove the resistance effect, biochemical analyses were performed to quantify proteins and the enzymatic activity of beta-1,3 glucanase. Results demonstrated that barley plants treated with the inducer, showed an increase in the concentration of proteins, as well as in the activity of the enzyme beta-1,3 glucanase, when compared with the extract from healthy plants. In infected plants, protein concentrations decreased and enzymatic activity was lower than in healthy plants. Results suggest that barley plants treated with xanthan gum developed mechanisms responsible for induced resistance, which are still unknown. The most important macromolecule in the defense mechanism was demonstrated to be PR-protein, due to its accumulation and concentration of proteins. However, it may not be the only macromolecule responsible for the resistance effect. (C) 2004 Elsevier SAS. All rights reserved.

Combinatorial synthesis and directed evolution applied to the production of alpha-helix forming antimicrobial peptides analogues

Antimicrobial peptides (AMPs) are effector molecules of innate immune systems found in different groups of organisms, including microorganisms, plants, insects, amphibians and humans. These peptides exhibit several structural motifs but the most abundant AMPs assume an amphipathic alpha-helical structure. The alpha-helix forming antimicrobial peptides are excellent candidates for protein engineering leading to an optimization of their biological activity and target specificity. Nowadays several approaches are available and this review deals with the use of combinatorial synthesis and directed evolution in order to provide a high-throughput source of antimicrobial peptides analogues with enhanced lytic activity and specificity.

Production and characterization of alpha-amylase from Rhizopus sp

A strain of Rhizopus sp. screened among more than 800 filamentous fungi showed great ability to produce a thermostable alpha-amylase by solid state fermentation. The best production was obtained with a bran moisture content of 40% when the enzyme activity reached 60 EU/g. of medium. During the purification procedures, a column of DEAE-Sephadex A-50 separated the enzyme in two fractions and the larger (85% of the total activity) showed optimum pH in a range from 4.0 to 5.6. Optimum temperature was found at 60-65 degrees C and in this range no loss of activity was observed after 60 min. of treatment in pH 5,0. Its K-m and V-m are, respectively, of 5.0 mg/ml of starch and 10,01 uMol of reducing sugar/min./mg. of protein. Its molecular weight was calculated in 64.000 by gel filtration in Sephadex G-200. The dextrinization power of the enzyme was observed preferentialy on substrates compound by chains with higher ramifications, that is: amylopectin > starch > amylose. Other aspects of the enzyme pattern action are also discussed.

Genetic parameters estimate for milk and mozzarella cheese yield, fat and protein percentage in dairy buffaloes in Brazil

The aim of this study was analyze the (co)variance components and genetic and phenotypic relationships in the following traits: accumulated milk yield at 270 days (MY270,), observed until 305 days of lactation; accumulated milk yield at 270 days (MY270/A) and at 305 days (MY305), observed until 335 days of lactation; mozzarella cheese yield (MCY) and fat (FP) and protein (PP) percentage, observed until 335 days of lactation. The (co)variance components were estimated by Restricted Maximum Likelihood methodology in analyses single, two and three-traits using animal models. Heritability estimated for MY270, MY270/A, MY305, MCY, FP and PP were 0.22; 0.24, 0.25, 0.14, 0.29 and 0.40 respectively. The genetic correlations between MCY and the variables MY270, MY270/A, MY305, PP and FP was: 0.85; 1.00; 0.89; 0.14 and 0.06, respectively. This way, the selection for the production of milk in long period should increase MCY. However, in the search of animals that produce milk with quality, the genetic parameters suggest that another index should be composed allying these studied traits.

Biomass production and secretion of hydrolytic enzymes are influenced by the structural complexity of the nitrogen source in Fusarium oxysporum and Aspergillus nidulans

The structural complexity of the nitrogen sources strongly affects biomass production and secretion of hydrolytic enzymes in filamentous fungi. Fusarium oxysporum and Aspergillus nidulans were grown in media containing glucose or starch, and supplemented with a nitrogen source varying from a single ammonium salt (ammonium sulfate) to free amino acids (casamino acids), peptides (peptone) and protein (gelatin). In glucose, when the initial pH was adjusted to 5.0, for both microorganisms, higher biomass production occurred upon supplementation with a nitrogen source in the peptide form (peptone and gelatin). With a close to neutrality pH, biomass accumulation was lower only in the presence of the ammonium salt. When grown in starch, biomass accumulation and secretion of hydrolytic enzymes (amylolytic and proteolytic) by Fusarium also depended on the nature of the nitrogen supplement and the pH. When the initial pH was adjusted to 5.0, higher growth and higher amylolytic activities were detected in the media supplemented with peptone, gelatin and casamino acids. However, at pH 7.0, higher biomass accumulation and higher amylolytic activities were observed upon supplementation with peptone or gelatin. Ammonium sulfate and casamino acids induced a lower production of biomass, and a different level of amylolytic enzyme secretion: high in ammonium sulfate and low in casamino acids. Secretion of proteolytic activity was always higher in the media supplemented with peptone and gelatin. Aspergillus, when grown in starch, was not as dependent as Fusarium on the nature of nitrogen source or the pH. The results described in this work indicate that the metabolism of fungi is regulated not only by pH, but also by the level of structural complexity of the nitrogen source in correlation to the carbon source.

Reduction of erythroid progenitors in protein-energy malnutrition

Protein-energy malnutrition is a syndrome in which anaemia together with multivitamin and mineral deficiency may be present. The pathophysiological mechanisms involved have not, however, yet been completely elucidated. The aim of the present study was to evaluate the pathophysiological processes that occur in this anaemia in animals that were submitted to protein-energy malnutrition, in particular with respect to Fe concentration and the proliferative activity of haemopoietic cells. For this, histological, histochemical, cell culture and immunophenotyping techniques were used. Two-month-old male Swiss mice were submitted to protein-energy malnutrition with a low-protein diet (20g/kg) compared with control diet (400 g/kg). When the experimental group had attained a 20% loss of their original body weight, the animals from both groups received, intravenously, 20IU erythropoietin every other day for 14 d. Malnourished animals showed a decrease in red blood cells, Hb concentration and reticulocytopenia, as well as severe bone marrow and splenic atrophy. The results for serum Fe, total Fe-binding capacity, transferrin and erythropoietin in malnourished animals were no different from those of the control animals. Fe reserves in the spleen, liver and bone marrow were found to be greater in the malnourished animals. The mixed colony-forming unit assays revealed a smaller production of granulocyte-macrophage colony-forming units, erythroid burst-forming units, erythroid colony-forming units and CD45, CD117, CD119 and CD71 expression in the bone marrow and spleen cells of malnourished animals. These findings suggest that, in this protein-energy malnutrition model, anaemia is not caused by Fe deficiency or erythropoietin deficiency, but is a result of ineffective erythropoiesis.

Effects of dietary macronutrient content on energy metabolism and uncoupling protein mRNA expression in broiler chickens

The objective of the present study was to investigate the effects of dietary macronutrient ratio on energy metabolism and on skeletal muscle mRNA expression of avian uncoupling protein (UCP), thought to be implicated in thermogenesis in birds. Broiler chickens from 2 to 6 weeks of age received one of three isoenergetic diets containing different macronutrient ratios (low-lipid (LL) 30 v. 77 g lipid/kg-, low-protein (LP) 125 v. 197 g crude protein (N X 6.25)/kg; low-carbohydrate (LC) 440 v. 520 g carbohydrate/kg). LP chickens were characterised by significantly lower body weights and food intakes compared with LL and LC chickens (-47 and -38% respectively) but similar heat production/kg metabolic body weight, as measured by indirect calorimetry, in the three groups. However, heat production/g food ingested was higher in animals receiving the LP diet (+41%, P<0.05). These chickens also deposited 57% less energy as protein (P<0.05) and 33% more as fat. No significant differences in energy and N balances were detected between LL and LC chickens. The diets with the higher fat contents (i.e. The LP and LC diets) induced slightly but significantly higher relative expressions of avian UCP mRNA in gastrocnemius muscle, measured by reverse transcription-polymerase chain reaction, than the LL diet (88 and 90 v. 78% glyceraldehyde-3-phosphate dehydrogenase respectively, P<0.05). Our present results are consistent with the recent view that UCP homologues could be involved in the regulation of lipid utilisation as fuel substrate and provide evidence that the macronutrient content of the diet regulates energy metabolism and especially protein and fat deposition.

Net energy, protein and macrominerals requirements for 70 to 120 day old female rabbits

In order to determine the net energy, protein and macrominerals requirements of 70 to 120 day old, 52 female White New Zealand rabbits, weighing 1900g +/- 40g were used. At the beginning of the experimental period, 14 of the 52 young does were slaughtered and the 38 remaining animals were kept under two dietary management: ad libitum and restricted feeding. Slaughters were performed to determine each nutrient body content. The weight gain nutrient requirements depicted by the quantities of each nutrient stored into the body were obtained by applying the regression equation, which estimate the empty body nutrient content logarithm as a function of the empty body weight logarithm, as described by ARC (1980). By determining the heat production logarithm at the zero level of metabolizable energy intake, the maintenance net energy requirement was estimated to be 45.31 Kcal/day/Kg(0.75) the mean net energy. protein, calcium, phosphorous, sodium, magnesium and potassium requirements for each gram of weight gain per day were estimated to be, 2.51 Kcal, 0.21g, 0.02g, 0.005g, 0.001g, 0.0004g and 0.002g, respectively.

Single dose of a vaccine based on DNA encoding mycobacterial hsp65 protein plus TDM-loaded PLGA microspheres protects mice against a virulent strain of Mycobacterium tuberculosis

The high incidence of tuberculosis around the world and the inability of BCG to protect certain populations clearly indicate that an improved vaccine against tuberculosis is needed. A single antigen, the mycobacterial heat shock protein hsp65, is sufficient to protect BALB/c mice against challenge infection when administered as DNA vaccine in a three-dose-based schedule. In order to simplify the vaccination schedule, we coencapsulated hsp65-DNA and trehalose dimicolate (TDM) into biodegradable poly(DL-lactide-co-glycolide) (PLGA) microspheres. BALB/c mice immunized with a single dose of DNA-hsp65/TDM-1oaded microspheres produced high levels of IgG2a subtype antibody and high amounts of IFN-gamma in the supernatant of spleen cell cultures. DNA-hsp65/TDM-loaded microspheres were also able to induce high IFN-gamma production in bulk lung cells from challenged mice and confer protection as effective as that attained after three doses of naked DNA administration. This new formulation also allowed a ten-fold reduction in the DNA dose when compared to naked DNA. Thus, this combination of DNA vaccine and adjuvants with immunomodulatory and carrier properties holds the potential for an improved vaccine against tuberculosis.

Energy and protein requirements for maintenance and growth of Boer crossbred kids

Meat production by goats has become an important livestock enterprise in several parts of the world. Nonetheless, energy and protein requirements of meat goats have not been defined thoroughly. The objective of this study was to determine the energy and protein requirements for maintenance and growth of 34 3/4 Boer x 1/4 Saanen crossbred, intact male kids (20.5 +/- 0.24 kg of initial BW). The baseline group was 7 randomly selected kids, averaging 21.2 +/- 0.36 kg of BW. An intermediate group consisted of 6 randomly selected kids, fed for ad libitum intake, that were slaughtered when they reached an average BW of 28.2 +/- 0.39 kg. The remaining kids (n = 21) were allocated randomly on d 0 to 3 levels of DMI (treatments were ad libitum or restricted to 70 or 40% of the ad libitum intake) within 7 slaughter groups. A slaughter group contained 1 kid from each treatment, and kids were slaughtered when the ad libitum treatment kid reached 35 kg of BW. Individual body components (head plus feet, hide, internal organs plus blood, and carcass) were weighed, ground, mixed, and subsampled for chemical analyses. Initial body composition was determined using equations developed from the composition of the baseline kids. The calculated daily maintenance requirement for NE was 77.3 +/- 1.05 kcal/kg(0.75) of empty BW (EBW) or 67.4 +/- 1.04 kcal/kg(0.75) of shrunk BW. The daily ME requirement for maintenance (118.1 kcal/g(0.75) of EBW or 103.0 kcal/kg(0.75) of shrunk BW) was calculated by iteration, assuming that the heat produced was equal to the ME intake at maintenance. The partial efficiency of use of ME for NE below maintenance was 0.65. A value of 2.44 +/- 0.4 g of net protein/kg(0.75) of EBW for daily maintenance was determined. Net energy requirements for growth ranged from 2.55 to 3.0 Mcal/kg of EBW gain at 20 and 35 kg of BW, and net protein requirements for growth ranged from 178.8 to 185.2 g/kg of EBW gain. These results suggest that NE and net protein requirements for growing meat goats exceed the requirements previously published for dairy goats. Moreover, results from this study suggest that the N requirement for maintenance for growing goats is greater than the established recommendations.

Evaluation of in vitro gas production of roughages and their mixtures using the curves subtraction method

The in vitro gas production of four single roughages and their paired combinations (1:1 on dry matter basis) were evaluated. Two roughage samples (100 mg) per treatment were fermented with ruminal fluid during a 48 h incubation period. Total 48 h gas volumes of fermentation dry matter (DM), neutral detergent fiber (NDF) and soluble compounds in neutral detergent (NDS) were for sugarcane = 16.8, 11.2, 6.9 mL; sugarcane + corn silage = 20.1, 12.6, 9.1 mL; sugarcane + 60-day elephantgrass = 16.5, 17.6 mL; sugarcane + 180-day elephantgrass = 13.8, 8.2, 5.9 mL; corn silage = 18.8, 16.8, 4.7 mL; corn silage + 60-day elephantgrass = 16.3, 15.4, 2.4 mL; corn silage + 180-day elephantgrass = 16.1, 11.8, 4.2 mL; 60-day elephantgrass = 16.9, 19.0 mL and 180-day elephantgrass = fermented 10.7, 12.2 mL, respectively. The NDS gas production was not possible to estimate for sugarcane + 60-day elephantgrass, 60-day elephantgrass and 180-day elephantgrass. The present data shows that the curves subtraction method can be an option to evaluate the contribution of the soluble fractions in roughages to digestion kinetics. However, this method underestimates the NDS gas contribution when roughages are low in crude protein and soluble carbohydrates. It is advisable to directly apply the two-compartmental mathematical model to the digestion curves for roughage DM, when determining the NDS gas volume and the digestion rate. This method is more straightforward and accurate when compared to the curve subtraction method. Non-structural carbohydrates combined with fiber and protein promoted a positive associative effect in sugarcane + corn silage (50:50) mixture. Therefore, it can be concluded that the soluble fraction of roughages greatly contributes to gas production. (C) 2004 Elsevier B.V. All rights reserved.

Macrophage suppression following phagocytosis of apoptotic neutrophils is mediated by the S100A9 calcium-binding protein

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