Protein diets promote the maturation of oocytes and spawning of Piaractus mesopotamicus kept in cages

The objective of this study was to evaluate the effects of balanced diets on the maturation of oocytes and the reproductive performance of P. mesopotamicus in cages. A completely random design with 224 fish in 16 cages measuring 5 m(3) was employed for this purpose. The treatments consisted of diets containing 18, 24, 30, and 36% crude protein (CP) provided ad libitum. The external and internal morphological characteristics of the specimens were examined, as well as: the position of the germinal vesicle, the distribution of oocyte diameters, the fertilization and hatching rates, the number of oocytes released, the total number of oocytes, the remaining weight and total weight of the ovaries, the gonadosomatic index, the condition factor (K), and the histology of the oocytes and ovaries post-spawning and during ovarian regression. The diameters of the oocytes collected before the first hormonal application displayed a unimodal distribution for the lowest protein content and a polymodal distribution for the other treatments. A similar situation was seen during spawning. The lowest fertilization and hatching rates were found as a consequence of the treatment with 30% CP (P < 0.05). The greatest hatching rate occurred in the females fed 18% CP. The greatest total oocyte weight was found in the specimens that received between 30 and 36% CP. The lowest K index was found in the females fed 36% CP. In conclusion, a diet containing 18% CP satisfies the reproductive requirements of females adapted to this system.

Histochemistry and protein profile of the mandibular glands of workers of the ant Atta sexdens rubropilosa (Hymenoptera: Formicidae)

Insect mandibular glands are always associated to the mandibles; they are part of the salivary glandular system. The mandibular glands are composed by a reservoir associated to the secretory cells, with each secretory cell connected to the reservoir by means of individual canaliculi. These glands play an important role in the production of pheromones, which are compounds involved in defense, communication, and reproduction of the colony. Mandibular glands of soldiers and major and minor workers of the ant Atta sexdens rubropilosa were processed for different histochemical tests, total protein content, and protein electrophoretic profile determination. The histochemical tests detected the presence of lipids, DNA/RNA, polysaccharides, and proteins at different regions of the gland. The protein electrophoretic profiles showed that the total protein content as well as the number of peptides of each caste follow a progressive order in relation to the size of the individual. Thus, we suggest that the production of secretion is directly linked to the task that the individual performs in the colony.

A comparative study of protein and enzymatic activity in venoms of some common wasps (Hymenoptera: Vespidae) from São Paulo State

The Hymenoptera Aculeata venoms, with few exceptions, have been poorly studied and characterized. Nevertheless, they have raised increasing interest due to their medical importance, since accidents with these insects are fairly frequent in Brazil and may cause severe allergic reactions. The objectives of the present work were the quantitative characterization of the main allergenic enzymes present in the venom of the species Polybia paulista, Polybia ignobilis, Polistes simillimus, and Agelaia pallipes pallipes through biochemical assays for the determination of total protein content, as well as the level of the enzymatic activity of phospholipase, hyaluronidase, acid phosphatase and esterase. These results, in addition to providing biochemical knowledge about the venom of the species in question, also supply studies that allow phylogenetic inferences among them.

Morphology and protein patterns of honey bee drone accessory glands.

We used light and transmission electron microscopy to examine the morphology of the accessory glands of immature and mature adult males of Apis mellifera L. We also made an electrophoretic analysis of the protein content of the mature gland. The glands of the immature male actively secrete a mucous substance that can be seen in the lumen of the gland of the mature male. This secretion stains with mercury bromophenol blue and with periodic acid-Schiff reaction, which stain glyconjugates. The protein content was higher in the lumen secretion than in the gland wall extracts. The electrophoresis patterns of the wall extracts were different from those of the secretion found in the gland lumen.

Influence of protein intake on weight loss in obese cats

The effects of two hypocaloric diets were evaluated, one with 29% and the other with 42% crude protein, on the body composition, nitrogen balance (NB), and some biochemical parameters of obese cats. A total of 16 castrated adult cats were used and divided into two groups of eight animals each. Body composition, determined by dual-energy x-ray absortiometry scanning, and biochemical examinations, were performed at the onset of the experiment (M0), at 10% of weight loss (M10), and at 20% of weight loss (M20) for each cat. The weekly weight loss (0.98 ± 0.37% for group 1; 0.94 ± 0.31% for group 2) and the ingestion of metabolizable energy (33.7 ±3.3 kcal/kg/day for group 1; 35.1 ±3.20 kcal/kg/day for group 2) did not differ between the groups. The NB was different at M0 (-70 ±110 mg/kg/day for group 1 ; 340 ±110 mg/kg/day for group 2) but roughly similar at M20 (140 ±170 mg/kg/day for group 1; 330 ± 410 mg/kg/day for group 2). The lean body mass (LM) loss was significant for group 1 (P < .05) in that it decreased from 2.789 ±198 g at M0 to 2.563 ±188 g at M20; for group 2, the changes in LM were not significant (P > .05). Reduction in body fat was significant between M0 and M20 for both diets (P < .05), without differences between treatments. The ingestion of digestible protein was greater (P < .05) for group 2 (3.20 ±0.29 g/kg/day) than for group 1 (2.21 ± 0.22 g/kg/day). There was a significant correlation between NB and ingestion of digestible protein at M0 (P < .05; r = 0.65), but this correlation was not observed at M20 (P > .05; r = 0.31). A significant reduction in plasma urea was observed for group 1 and in high-density lipoprotein cholesterol for group 2, but the other biochemical parameters did not change. The diet with higher protein content prevented LM loss. However, the lower-protein diet seems to maintain animal health and improve the cats' NB after weight loss.

Decreased rate of protein synthesis, caspase-3 activity, and ubiquitin-proteasome proteolysis in soleus muscles from growing rats fed a low-protein, high-carbohydrate diet

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

High protein microparticles produced by ionic gelation containing Lactobacillus acidophilus for feeding pacu larvae

Microparticles with high protein content can be used as diets to mimic the proximate composition of Artemia nauplii. After production, the particles were characterized with respect to their proximate composition, mean size, morphology, and rehydration behavior after drying. The protein content, lipid content and the particle moisture were similar to Artemia nauplii, with mean values of 50, 23, and 85%, respectively. Additionally, the particles were used in a pacu (Piaractus mesopotamicus) larval growth experiment. Also, the probiotic Lactobacillus acidophilus was added to one of the diets, and the effects of the diets were evaluated on larvae growth and stress resistance. Larvae fed the experimental diets had lower growth than larvae fed with Artemia nauplii or a commercial diet. All of the evaluated diets, including the experimental ones, showed high ingestion rates (>90%). In the stress test by air exposure, larvae fed with the microparticle without probiotic exhibited a significantly higher mortality than those fed the commercial diet or those fed with Artemia nauplii. The low growth rates may have been due to a potential nutritional inadequacy with respect to the low mineral/vitamin content of the experimental diets. (C) 2014 Elsevier Ltd. All rights reserved.

Metabolism and ruminal parameters of Holstein × Gir heifers fed sugarcane and increasing levels of crude protein

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Prediction of culture performance of juvenile Litopenaeus vannamei by in vitro (pH-stat) degree of feed protein hydrolysis with species-specific enzymes

Rapid in vitro methods for measuring digestibility may be useful in analysing aqua feeds if the extent and limits of their application are clearly defined. The pH-stat protein digestibility routine with shrimp hepatopancreas enzymes was previously related to apparent protein digestibility with juvenile Litopenaeus vannamei fed diets containing different protein ingredients. The potential of the method to predict culture performance of shrimp fed six commercial feeds (T3, T4, T5, T6, T7 and T8) with 350 g kg(-1) declared crude-protein content was assessed. The consistency of results obtained using hepatopancreas enzyme extracts from either pond or clear water-raised shrimp was further verified in terms of reproducibility and possible diet history effects upon in vitro outputs. Shrimps were previously acclimated and then maintained over 56 days (initial mean weight 3.28 g) on each diet in 500-L tanks at 114 ind m(-2), clear water closed system with continuous renewal and mechanical filtering (50 mu m), with four replicates per treatment. Feeds were offered four times daily (six days a week) delivered in trays at feeding rates ranging from 4.0% to 7.0% of stocked shrimp biomass. Feed was accessible to shrimp 4 h daily for 1-h feeding period after which uneaten feed was recovered. Growth and survival were determined every 14 days from a sample of 16 individuals per tank. Water quality was monitored daily (pH, temperature and salinity) and managed by water back flushing filter cleaning every 7-10 days. Feeds were analysed for crude protein, gross energy, amino acids and pepsin digestibility. In vitro pH-stat degree of protein hydrolysis (DH%) was determined for each feed using hepatopancreas enzyme extracts from experimental (clear water) or pond-raised shrimp. Feeds resulted in significant differences in shrimp performance (P < 0.05) as seen by the differences in growth rates (0.56-0.98 g week(-1)), final weight and feed conversion ratio (FCR). Shrimp performance and in vitro DH% with pond-raised shrimp enzymes showed significant correlation (P < 0.05) for yield (R-2 = 0.72), growth rates (R-2 = 0.72-0.80) and FCR (R-2 = -0.67). Other feed attributes (protein : energy ratio, amino acids, true protein, non-protein nitrogen contents and in vitro pepsin digestibility) showed none or limited correlation with shrimp culture performance. Additional correlations were found between growth rates and methionine (R-2 = 0.73), FCR and histidine (R-2 = -0.60), and DH% and methionine or methionine+cystine feed contents (R-2 = 0.67-0.92). pH-stat assays with shrimp enzymes generated reproducible DH% results with either pond (CV <= 6.5%) or clear water (CV <= 8.5%) hepatopancreas enzyme sources. Moreover, correlations between shrimp growth rates and feed DH% were significant regardless of the enzyme origin (pond or clear water-raised shrimp) and showed consistent R-2 values. Results suggest the feasibility of using standardized hepatopancreas enzyme extracts for in vitro protein digestibility.

Protein diets promote the maturation of oocytes and spawning of Piaractus mesopotamicus kept in cages

The objective of this study was to evaluate the effects of balanced diets on the maturation of oocytes and the reproductive performance of P. mesopotamicus in cages. A completely random design with 224 fish in 16 cages measuring 5 m(3) was employed for this purpose. The treatments consisted of diets containing 18, 24, 30, and 36% crude protein (CP) provided ad libitum. The external and internal morphological characteristics of the specimens were examined, as well as: the position of the germinal vesicle, the distribution of oocyte diameters, the fertilization and hatching rates, the number of oocytes released, the total number of oocytes, the remaining weight and total weight of the ovaries, the gonadosomatic index, the condition factor (K), and the histology of the oocytes and ovaries post-spawning and during ovarian regression. The diameters of the oocytes collected before the first hormonal application displayed a unimodal distribution for the lowest protein content and a polymodal distribution for the other treatments. A similar situation was seen during spawning. The lowest fertilization and hatching rates were found as a consequence of the treatment with 30% CP (P < 0.05). The greatest hatching rate occurred in the females fed 18% CP. The greatest total oocyte weight was found in the specimens that received between 30 and 36% CP. The lowest K index was found in the females fed 36% CP. In conclusion, a diet containing 18% CP satisfies the reproductive requirements of females adapted to this system.

Thermal studies on protein isolates of white lupin seeds (Lupinus albus)

This study used TG, DSC, and SDS-PAGE techniques to study protein isolates (PIs) in the powder form obtained from lupin seeds flour Lupinus albus. Different methods of preparing PIs were tested, resulting in final products that were different only in relation to the yield and protein content. The results of the protein analysis by SDS-PAGE showed that the same protein fractions were present in the lupin seeds and in the obtained PIs. This result shows that the process of extraction was not damaging to the composition of the original protein. On the other hand, the results of the thermal analysis (DSC and TG-DTG curves) obtained for the different PIs, led to the detection of changes in the protein conformation through the Delta H values, which in general decreased with increasing values of pH and ionic strength in the experimental conditions of extraction.

Inhibition of cannabinoid CB1 receptor upregulates Slc2a4 expression via nuclear factor-kappa B and sterol regulatory element-binding protein-1 in adipocytes

Evidences have suggested that the endocannabinoid system is overactive in obesity, resulting in enhanced endocannabinoid levels in both circulation and visceral adipose tissue. The blockade of cannabinoid receptor type 1 (CB1) has been proposed for the treatment of obesity. Besides loss of body weight, CB1 antagonism improves insulin sensitivity, in which the glucose transporter type 4 (GLUT4) plays a key role. The aim of this study was to investigate the modulation of GLUT4-encoded gene (Slc2a4 gene) expression by CB1 receptor. For this, 3T3-L1 adipocytes were incubated in the presence of a highly selective CB1 receptor agonist (1 mu M arachidonyl-2'-chloroethylamide) and/or a CB1 receptor antagonist/inverse agonist (0.1, 0.5, or 1 mu M AM251, 1-(2,4-dichlorophenyl)-5-(4-iodophenyl)-4-methyl-N-1-piperidinyl-1H-pyrazole-3-carboxamide). After acute (2 and 4 h) and chronic (24 h) treatments, cells were harvested to evaluate: i) Slc2a4, Cnr1 (CB1 receptor-encoded gene), and Srebf1 type a (SREBP-1a type-encoded gene) mRNAs (real-time PCR); ii) GLUT4 protein (western blotting); and iii) binding activity of nuclear factor (NF)-kappa B and sterol regulatory element-binding protein (SREBP)-1 specifically in the promoter of Slc2a4 gene (electrophoretic mobility shift assay). Results revealed that both acute and chronic CB1 receptor antagonism greatly increased (similar to 2.5-fold) Slc2a4 mRNA and protein content. Additionally, CB1-induced upregulation of Slc2a4 was accompanied by decreased binding activity of NF-kappa B at 2 and 24 h, and by increased binding activity of the SREBP-1 at 24 h. In conclusion, these findings reveal that the blockade of CB1 receptor markedly increases Slc2a4/GLUT4 expression in adipocytes, a feature that involves NF-kappa B and SREBP-1 transcriptional regulation. Journal of Molecular Endocrinology (2012) 49, 97-106

Development of corn and flaxseed snacks with high-fibre content using response surface methodology (RSM)

With the increasing emphasis on health and well-being, nutrition aspects need to be incorporated as a dimension of product development. Thus, the production of a high-fibre content snack food from a mixture of corn and flaxseed flours was optimized by response surface methodology. The independent variables considered in this study were: feed moisture, process temperature and flaxseed flour addition, as they were found to significantly impact the resultant product. These variables were studied according to a rotatable composite design matrix (-1.68, -1, 0, 1, 1.68). Response variable was the expansion ratio since it has been highly correlated with acceptability. The optimum corn-flaxseed snack obtained presented a sevenfold increase in dietary fibre, almost 100% increase in protein content compared to the pure corn snack, and yielded an acceptability score of 6.93. This acceptability score was similar to those observed for corn snack brands in the market, indicating the potential commercial use of this new product, which can help to increase the daily consumption of dietary fibre.

Palmitoleic acid (n-7) increases white adipocyte lipolysis and lipase content in a PPARα-dependent manner

We investigated whether palmitoleic acid, a fatty acid that enhances whole body glucose disposal and suppresses hepatic steatosis, modulates triacylglycerol (TAG) metabolism in adipocytes. For this, both differentiated 3T3-L1 cells treated with either palmitoleic acid (16:1n7, 200 μM) or palmitic acid (16:0, 200 μM) for 24 h and primary adipocytes from wild-type or PPARα-deficient mice treated with 16:1n7 (300 mg•kg(-1)•day(-1)) or oleic acid (18:1n9, 300 mg•kg(-1)•day(-1)) by gavage for 10 days were evaluated for lipolysis, TAG, and glycerol 3-phosphate synthesis and gene and protein expression profile. Treatment of differentiated 3T3-L1 cells with 16:1n7, but not 16:0, increased basal and isoproterenol-stimulated lipolysis, mRNA levels of adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL) and protein content of ATGL and pSer(660)-HSL. Such increase in lipolysis induced by 16:1n7, which can be prevented by pharmacological inhibition of PPARα, was associated with higher rates of PPARα binding to DNA. In contrast to lipolysis, both 16:1n7 and 16:0 increased fatty acid incorporation into TAG and glycerol 3-phosphate synthesis from glucose without affecting glyceroneogenesis and glycerokinase expression. Corroborating in vitro findings, treatment of wild-type but not PPARα-deficient mice with 16:1n7 increased primary adipocyte basal and stimulated lipolysis and ATGL and HSL mRNA levels. In contrast to lipolysis, however, 16:1n7 treatment increased fatty acid incorporation into TAG and glycerol 3-phosphate synthesis from glucose in both wild-type and PPARα-deficient mice. In conclusion, palmitoleic acid increases adipocyte lipolysis and lipases by a mechanism that requires a functional PPARα

Histopathological alterations, EROD activity, CYP1A protein and biliary metabolites in gilthead seabream Sparus aurata exposed to benzo(a)pyrene

This study compared for seabream, Sparus aurata exposed to benzo(a)pyrene-B(a)P-, the response of molecular cytochrome P450 1A (CYP1A) and cellular histopathology biomarkers. Male gilthead seabream, Sparus aurata specimens were exposed for 20 days via water to a series of high B(a)P concentrations. CYP1A was assessed by measuring enzymatic activity (EROD) and CYP1A protein content, and cellular responses were evaluated by routine histopathological methods. In addition, biliary metabolites were measured in order to verify that B(a)P was absorbed and metabolised. Histological lesions, both in liver and gills, increased in parallel to B(a)P concentrations, with the majority of changes representing rather non-specific alterations. Hepatic EROD and CYP1A proteins data showed a concentration-dependent induction, while in the gills, EROD activity but not CYP1A proteins showed a non-monotonous dose response, with a maximum induction level at 200 microg B(a)P.L-1 and decreasing levels thereafter. The findings provide evidence that short-term, high dose exposure of fish can result in significant uptake and metabolism of the lipophilic B(a)P, and in pronounced pathological damage of absorptive epithelia and internal organs.