Marine microalgae as a potential source of single cell protein (SCP)

Summary The marine microalgaeTetraselmis suecica, Isochrysis galbana, Dunaliella tertiolecta andChlorella stigmatophora are good biological sources of single cell protein (SCP). Protein content accounts for 39.12%–54.20% of the dry matter,D. tertiolecta having the highest. Lysine values are between 3.67 and 4.52 g/100 g of protein, and thus are higher than those for freshwater species. The total nucleic acid content is less than 7% of the dry matter; this value is definitely lower than that for yeasts or bacteria, commonly used as SCP sources. Amino acid profiles of the four species are very similar and comparable to the FAO reference protein, buth with a low content of methionine and cystine and a high content of lysine. The MEAA indices are between 81 and 84.98, without significant differences among the four species. Marine microalgae can be used as a potential SCP source.     

Effects of alginates and sodium carrageenates mixed with soy proteins on the coefficient of protein efficiency

Male 21 d-old Wistar rats, were fed for 4 wk with diets containing casein or soybean proteins (10%) with 0.5, 1, 2 or 3% sodium alginate or sodium carrageenan or without any alginate or carrageenan. Daily protein intake and weight gain of casein-fed rats were not significantly different (P less than 0.05) from those of rats fed soybean meal with alginate, whatever the dose received. Rats fed 3% carrageenan in soybean meal had significantly higher feed intake than that of rats fed casein. At the levels studied, alginate had no effect on the Protein Efficiency Ratio (PER), but carrageenan did. The addition of increased quantities of carrageenan to soybean meal followed by heating the mixture led to a progressive and significant decrease in PER at all levels of carrageenan compared to casein feeding. The addition of 3% carrageenan to heated soybean meal, corresponding to 0.62% of meal diet, led to a significant decrease in PER. These results confirm the precipitating role of carrageenans on proteins.     

Protein utilization in correlation to protein intake

In a 14-day experiment, weaned and adult rats were given ad libitum isocaloric diets with a mounting casein content (5, 10, 15, 25 and 40% by weight) and growth parameters of protein biological value, PER and NPR, and the utilization parameters NPU (body protein) and LPU (liver protein) were determined together with phosphoenolpyruvate carboxykinase (gluconeogenetic enzyme) and pyruvate kinase (glycolytic enzyme) activity in the animals' liver. The decrease in all the biological value parameters in weaned rats on 25% and 40% casein diets and in adult rats on 15%, 25% and 40% casein diets shows that these concentrations are too high for the organism. The decrease in PER and diminished weight and body and liver nitrogen increments in both age groups in animals with a low protein intake is evidence that 5% casein is an inadequate concentration. The optimum diet for weaned rats is thus a 15% casein diet and for adult rats a 10% casein diet, as confirmed by the linear correlation between weight increments, body and liver nitrogen and protein intake and also by gluconeogenetic enzyme activity. Under the given experimental conditions the study is a contribution to the determination of optimum physiological doses of proteins.     

A Freshwater Clam (Corbicula fluminea) Extract Improves Cholesterol Metabolism in Rats Fed on a High-Cholesterol Diet

The effect of a freshwater clam (Corbicula fluminea) extract (FCE) on cholesterol metabolism in rats fed on a high-cholesterol diet was investigated. When rats were fed various amounts of FCE in addition to the high-cholesterol diet for 2 wk, the serum and hepatic cholesterol levels were gradually reduced in a dose-dependent manner, as compared with the control group. The excretion of neutral sterols and bile acids into the feces was increased by feeding FCE. Several phytosterols were detected in the feces of rats fed on the FCE-containing diet. In addition, substantial amounts of phytosterols were found in FCE. Cholesterol 7α-hydroxylase (CYP7A1) mRNA in the liver of the rats fed on the FCE-containing diets was higher than that of rats fed on the high-cholesterol diets without FCE. These results may suggest that enhanced cholesterol degradation and the excretion of neutral sterols and bile acids contributed to the hypocholesterolemic effect of FCE observed in the hypercholesterolemic rats fed on the high-cholesterol diet.     

Food restriction followed by refeeding with a casein- or whey-based diet differentially affects the gut microbiota of pre-pubertal male rats

Researchers are gaining an increasing understanding of host–gut microbiota interactions, but studies of the role of gut microbiota in linear growth are scarce. The aim of this study was to investigate the effect of food restriction and refeeding with different diets on gut microbiota composition in fast-growing rats. Young male Sprague–Dawley rats were fed regular rat chow ad libitum (control group) or subjected to 40% food restriction for 36 days followed by continued restriction or ad libitum refeeding for 24 days. Three different diets were used for refeeding: regular vegetarian protein chow or chow in which the sole source of protein was casein or whey. In the control group, the composition of the microbiota remained stable. Food restriction for 60 days led to a significant change in the gut microbiota at the phylum level, with a reduction in the abundance of Firmicutes and an increase in Bacteroidetes and Proteobacteria. Rats refed with the vegetarian protein diet had a different microbiota composition than rats refed the casein- or whey-based diet. Similarities in the bacterial population were found between rats refed vegetarian protein or a whey-based diet and control rats, and between rats refed a casein-based diet and rats on continued restriction. There was a significant strong correlation between the gut microbiota and growth parameters: humerus length, epiphyseal growth plate height, and levels of insulin-like growth factor 1 and leptin. In conclusion, the type of protein in the diet significantly affects the gut microbiota and, thereby, may affect animal's health.     

Distribution of Selenium in Bovine Milk and Selenium Deficiency in Rats Fed Casein-based Diets,Monitored by Lipid Peroxide Level and Glutathione Peroxidase Activity

A major proportion of selenium in bovine milk was found in fluorometric analysis to be associated with the casein fraction, largely in alkali-labile form, and the rest with the whey fraction mostly in free selenite form. This uneven distribution of milk selenium seems to provide an explanation for selenium deficiency in purified caseins. The activity of glutathione peroxidase, a selenoprotein, in the liver of growing male rats fed ad libitum low-selenium diets containing either vitamin-free casein or Torula yeast 0.065 ± 0.012 or 0.015 ± 0.004 μ g Se/g diet, respectively) for 3 weeks decreased to 4 to 6% of that of the control rats fed a commercial stock diet (0.185 ± 0.092 μ g Se/g diet). Selenium status was evaluated by three different parameters for the rats assigned under pair-feeding regimen to those vitamin-free casein-based diets which were supplemented with graded levels of selenium as sodium selenite. The hepatic levels of the thiobarbituric acid-reactive substance, an indication of lipid peroxidation, decreased to control level with selenium supplementation per g diet of 0.1 μ g and over. The hepatic glutathione peroxidase activity reached a plateau above a 0.1 μ g/g diet of selenium supplementation, whereas the erythrocyte enzyme activity increased with increasing levels of supplementary selenium. These results support the notion that semi-purified diets containing vitamin-free casein as a prime protein source would not satisfy the selenium requirement of growing animals unless deliberately supplemented with additional selenium.     

Conversion of Biotin Diaminocarboxylic Acid into Biotin and Bisnorbiotin by Resting Cell System of Microorganisms

The effects of dietary fat and protein levels on the conversion of Trp-Nam were investigated. In rats fed with 20% casein diets, the Trp-Nam conversion ratio [(urinary excretion of Nam + MNA + 2-Py + 4-Py in μmol/day)/(daily Trp intake during urine collection in μmol/day) × 100] was about 4.3% for the groups fed with the 20% corn oil and 20% soybean oil diets, 2.8% for the group fed with the 20% lard diet, and 2.1 % for the group fed with the no fat diet. In rats fed with 40% casein diets, a similar phenomenon was observed, but the ratios were 2.0%, 2.4%, 1.6%, and 0.8% for the groups fed with the 20% corn oil, 20% soybean oil, 20% lard, and no fat diets, respectively. From these results, it was found that an increase in fat intake elevated the conversion ratio regardless of the dietary protein level, while an increase in protein intake reduced it.     

Dietary arginine requirement of fingerling Indian major carp,Labeo rohita (Hamilton) based on growth,nutrient retention efficiencies,RNA/DNA ratio and body composition

To quantify the optimum dietary arginine requirement of fingerling Indian major carp, Labeo rohita (4.10 ± 0.04 cm; 0.62 ± 0.02 g), an 8‐week growth trial was conducted in eighteen 70‐L indoor circular aqua‐coloured troughs provided with a flow‐through system at 28 ± 1°C. Isonitrogenous (40 g 100 g^?1 crude protein) and isocaloric (4.28 kcal g^?1 gross energy) amino acid test diets containing casein and gelatin as intact protein sources with graded levels of arginine (0.5, 0.75, 1.0, 1.25, 1.50 and 1.75 g 100 g^?1 dry diet) were fed to triplicate groups of fish to apparent satiation at 07:00, 12:00 and 17:30 hours. Growth performance of fish fed the above diets was evaluated on the basis of absolute weight gain (AWG), specific growth rate (SGR), feed conversion ratio (FCR), protein efficiency ratio (PER), protein retention efficiency (PRE) and energy retention efficiency (ERE). Maximum AWG (2.61), SGR (2.80), best FCR (1.35), highest PER (1.85), PRE (37%) and ERE (76%) were recorded at 1.25 g 100 g^?1 dietary arginine. Maximum body protein (18.88 g 100 g^?1) and RNA/DNA ratio (5.20) were also obtained in a 1.25 g 100 g^?1 arginine dry diet. Except for the reduced growth performance in fish fed arginine‐deficient diets, no other deficiency signs were apparent. Based on the broken‐line and second‐degree polynomial regression analysis of the AWG, SGR, FCR, PER, PRE and ERE data, the optimum arginine requirement for fingerling Labeo rohita was found to be in the range of 1.22–1.39 g 100 g^?1 of the dry diet, corresponding to 3.05–3.47 g 100 g^?1 of dietary protein.     

A freshwater clam (Corbicula fluminea) extract improves cholesterol metabolism in rats fed on a high-cholesterol diet

The effect of a freshwater clam (Corbicula fluminea) extract (FCE) on cholesterol metabolism in rats fed on a high-cholesterol diet was investigated. When rats were fed various amounts of FCE in addition to the high-cholesterol diet for 2 wk, the serum and hepatic cholesterol levels were gradually reduced in a dose-dependent manner, as compared with the control group. The excretion of neutral sterols and bile acids into the feces was increased by feeding FCE. Several phytosterols were detected in the feces of rats fed on the FCE-containing diet. In addition, substantial amounts of phytosterols were found in FCE. Cholesterol 7alpha-hydroxylase (CYP7A1) mRNA in the liver of the rats fed on the FCE-containing diets was higher than that of rats fed on the high-cholesterol diets without FCE. These results may suggest that enhanced cholesterol degradation and the excretion of neutral sterols and bile acids contributed to the hypocholesterolemic effect of FCE observed in the hypercholesterolemic rats fed on the high-cholesterol diet.     

Cell growth of skeletal muscle as an indicator of protein requirements of the adult rat

Skeletal muscle growth, muscle nucleic acids and muscle protein synthesis capacity, were measured to evaluate the protein requirement of adult rats. Wistar rats were fed on diets containing 4%, 10% or 20% casein + D,L-methionine. All diets were provided for 21 days beginning at 90 days of age. Body weight, food efficiency and net weight change increased as the casein content of the diet increased. Muscle DNA, RNA and RNA/protein were lost, but protein and protein/DNA increased on the 4% and 20% protein diet. This fact involves an aplasia phenomenon although the hypertrophic growth is maintained. Alterations of the insulin and GH plasma levels were observed. These findings indicate that for adult rats the 4% and 20% protein diets are not adequate for the period of adult maintenance.     

High-Casein Diet Suppresses Guanidinoacetic Acid-Induced Hyperhomocysteinemia and Potentiates the Hypohomocysteinemic Effect of Serine in Rats

To determine the effect of dietary protein level on experimental hyperhomocysteinemia, rats were fed 10% casein (10C) and 40% casein (40C) diets with or without 0.5% guanidinoacetic acid (GAA) for 14 d. In addition, rats were fed 10C + 0.75% methionine (10CM) and 40C + 0.75% methionine (40CM) diets with or without 2.5% serine for 14 d to determine the relationship between the dietary protein level and intensity of the hypohomocysteinemic effect of serine. GAA supplementation markedly increased the plasma homocysteine concentration in rats fed with the 10C diet, whereas it did not increase the plasma homocysteine concentration in rats fed with the 40C diet. Although serine supplementation significantly suppressed the methionine-induced enhancement of plasma homocysteine concentration, the decreased plasma homocysteine concentration was significantly lower in rats fed with the 40CM diet than in rats fed with the 10CM diet. The hepatic cystathionine β-synthase and betaine-homocysteine S-methyltransferase activities were significantly higher in rats fed with the 40C or 40CM diet than in rats fed with the 10C or 10CM diet, irrespective of supplementation with GAA and serine. These results indicate that the high-casein diet was effective for both suppressing GAA-induced hyperhomocysteinemia and potentiating the hypohomocysteinemic effect of serine, probably through the enhanced activity of homocysteine-metabolizing enzymes.     

Changes in Creatine and Urea Formation in Rats Fasted and Fed Low Protein Diets

Changes in the time course of the urinary excretion of creatinine, creatine and urea, and the activities of kidney transamidinase and liver urea-cycle enzymes were investigated in rats fasted and fed on a 10% casein diet and 10% casein diets supplemented with 10% glycine and/or 1.4% arginine.

The urinary total-creatinine of the fasted rats increased extremely during fasting for 7 days, while that of the animals given the 10% casein diet supplemented with glycine and arginine rose exceedingly on the 3rd day and thereafter no significant change was observed. Most of the increase of total-creatinine could be accounted for by the increase of creatine. The activity of kidney transamidinase in the fasted rats decreased in the 3rd day and thereafter kept nearly constant. The transamidinase activity of rats fed on the 10% casein diet after giving a protein-free diet for 5 days increased in the 3rd day. An inverse relation was observed between the urinary creatine and the transamidinase activity. The urinary urea increased in the rats fasted or fed on the 10% casein diets with the supplement of glycine and/or arginine. In fasting, the activities of liver urea-cycle enzymes, except arginase, had a tendency of increasing with the lapse of time. The arginase activity remained more or less constant. The reason of the extreme increase of urinary creatine during starvation was discussed.     

Soy based diet attenuates the development of hypertension when compared to casein based diet in spontaneously hypertensive rat

The purpose of this study was to compare the effects of soy and casein based diets on blood pressure and cardiovascular functions in male and female spontaneously hypertensive rats (SHR). The systolic blood pressure was measured at the beginning and at the end of study. After a five week supplementation period with three different diets, the rats were decapitated and arterial responses and the weight-to-body weight-ratios of the organs were studied. The development of hypertension was attenuated in both female and male rats on soy protein diet when compared to the casein diet. Soy based diet lowered serum total cholesterol level when compared to the control diet. Both casein and soy protein supplementation in diet induced a significant renal hypertrophy in both female and male SHR rats when compared to SHR rats on the control diet. Soy protein supplementation reduced significantly serum estradiol-17beta concentration when compared to the control diet. There were no differences in the serum testosterone concentrations between the diet groups. When compared to the casein based diet the soy based diet attenuated the development of hypertension and decreased serum total cholesterol level in SHRs. These effects were independent of gender. The mechanisms and clinical importance of these findings remain to be clarified.     

Nutritional utilization of cottonseed flour, compared to soya flour, by growing rats with protein malnutrition

The aim of this study was to compare the nutritional value of three diets with rats submitted to protein malnutrition. After one month on a low protein diet (2% casein), the rats were refed with corn-soya-milk (60%/30%/5%, W/W/W) and rice-cotton-milk (60%/30%/5%) diets. This produced a higher growth rate than either the corn-cotton milk (60%/30%/5%) diets or a control diet containing 20% casein. The caloric intake/100 g body weight was about the same with the four rehabilitation diets. The highest apparent nitrogen digestive utilization coefficients (C. U. D.) values were obtained with the control diet, lower values with the rice-cotton-milk diet, then the corn-cotton-milk diet and lowest with the corn-soya-milk diet. Results of lipid C. U. D. were in the same order. The highest nitrogen balances were obtained with the control diet, then the corn-soya-milk diet, the rice-cotton-milk diet and lowest values with the corn-cotton-milk diet. Our data prove that the cotton flour mixed with rice may perfectly replace soya flour associated with corn in a nutritional rehabilitation diet.     

Supplementation of orotic acid to the casein, but not to egg protein, soy protein, or wheat gluten diets, increases serum ornithine carbamoyltransferase activity

Effects of dietary supplementation of orotic acid to a diet containing the casein protein were compared with diets containing egg protein, soy protein, or wheat gluten on lipid levels in the liver and serum and activities of ornithine carbamoyltransferase (OCT) and alanine aminotransferase in the serum of rats. We found that supplementation of orotic acid to each diet increased the contents of the liver total lipids, triacylglycerol, and phospholipids compared with those not supplemented. The contents of liver total lipids, triacylglycerol, cholesterol, and phospholipids in rats fed the casein diet were significantly higher than those of rats fed the other three diets when orotic acid was supplemented. The levels of triacylglycerol, cholesterol, and phospholipids in the serum of rats fed the casein diet were markedly decreased by addition of orotic acid. The supplementation of orotic acid significantly increased the activities of both serum OCT and alanine aminotransferase in rats fed the casein diet, but not in rats fed the other diets. In conclusion, liver lipid accumulation induced by dietary orotic acid depends on the type of dietary protein. The enhancement of serum OCT activity may result from liver lipid accumulation in rats fed the casein diet supplemented with orotic acid, demonstrating hepatic damage.     

Effects of protein type and concentration on development and reproduction of the German cockroach, Blattella germanica

Nymphal development and adult female reproduction were examined in the German cockroach, Blattella germanica, using a defined artificial diet in which the type of protein was varied. Milk proteins, including casein, supported development poorly compared to meat and plant proteins. Soybean protein supported development better than all other highly purified proteins including vitamin-free casein which is commonly used in artificial diets. Last instar females fed the soybean-based diet eclosed earlier at higher eclosion weights, developed their oocytes at a faster rate and experienced higher fecundity than females fed a vitamin-free casein-based diet. Last instar females exhibited different dose-response patterns on diets containing soybean isolate or vitamin-free casein. However, at all concentrations soybean protein was superior to casein in supporting development. The results of a food utilization study during the last instar revealed that consumption rates varied between females fed the soybean and casein based diets. However, approximate digestibility, efficiency of conversion of digested food and the efficiency of conversion of ingested food did not vary significantly between the two dietary treatments. Differential development of females fed the two diets was attributed to differences in stage-specific consumption rates and the poorer quality of casein as a source of protein for development in this species.     

Ingestion rates and dietary lipids affect growth and fatty acid composition of Dendraster excentricus larvae

This study investigated the effect of single and mixed algal diets on growth, fatty acid composition and ingestion rates for Dendraster excentricus larvae. Larvae were assigned to three single algal diet treatments Isochrysis galbana, Dunaliella tertiolecta or Rhodomonas sp. and four mixed algal diet treatments D. tertiolecta and Rhodomonas, I. galbana and D. tertiolecta, I. galbana and Rhodomonas, D. tertiolecta, Isochrysis galbana and Rhodomonas sp. Small amounts (0.36-0.6%) of stearic acid (18:0) were seen in the three algae used but a relatively large percentage (7-25%) of this SAFA was found in Dendraster larvae. The alga D. tertiolecta had the highest percentage (51.7%) of the short chain polyunsaturated fatty acid (PUFA) linolenic acid 18:3(n-3) and trace amounts (0.02-0.14%) of the long chain PUFAs eicosapentanoic (EPA, 20:5(n-3) and docosahexanoic acids (DHA, 22:6(n-3)). However, sand dollar larvae demonstrated the ability to elongate and desaturate shorter chain (18 carbon) polyunsaturated fatty acids (PUFA) to longer chain (20 carbon) n-3 PUFA. Thus high levels of 18:3(n-3) in D. tertiolecta led to high levels of EPA and low levels of 18:3(n-3) in Dendraster larvae fed this diet. Rhodomonas sp. had the highest percentage of stearidonic acid (18:4(n-3), 38.14%) and EPA (10.6%). Despite high levels of 18:4(n-3) in Rhodomonas sp. this acid was absent or found at very low levels in larvae fed this alga, or any combination of this alga. I. galbana had the highest percentage of DHA(14.3%) but was almost devoid of EPA (0.43%). Although Rhodomonas sp. and I. galbana had high levels of EPA and DHA sand dollar larvae did not incorporate higher levels of these long chain PUFAs into their lipids compared to those fed the alga D. tertiolecta. Dendraster larvae synthesized a number of 20 and 22 carbon non-methylene interrupted dienes (NMID), with levels increasing with larval stage. Higher ingestion rates were observed for Dendraster larvae fed single algal diets (Rhodomonas sp. or D. tertiolecta) and lower ingestion rates for those fed mixed algal diets. The highest ingestion rates were for 8-arm Dendraster larvae fed the large alga Rhodomonas sp. presented as a single algal diet. When fed a combination of three algae, selection of particles varied slightly depending on stage with 8-arm larvae ingesting slightly more of the larger algal cell in the mixture than 6-arm larvae. The present study suggests that regardless of the ratios given larvae might have an optimum ratio of different sized particles at which they can feed. The mixed algal diet of I. galbana and D. tertiolecta was the best algal diet leading to significantly larger larvae with high survival and development to metamorphosis. The single algal diets of Rhodomonas sp. or Dunaliella tertiolecta were the second and third best algal diets based on growth and survival to metamorphosis.     

Oleuropein supplementation increases urinary noradrenaline and testicular testosterone levels and decreases plasma corticosterone level in rats fed high-protein diet

The effects of oleuropein, a phenolic compound in extra virgin olive oil, on protein metabolism were investigated by measuring testicular testosterone and plasma corticosterone levels in rats fed diets with different protein levels. In Experiment 1, rats were fed experimental diets with different protein levels (40, 25 and 10 g/100 g casein) with or without 0.1 g/100 g oleuropein. After 28 days of feeding, the testosterone level in the testis was significantly higher and the plasma corticosterone level was significantly lower in rats fed the 40% casein diet with oleuropein than in those fed the same diet without oleuropein. The urinary noradrenaline level, nitrogen balance and hepatic arginase activity were significantly higher in rats fed the 40% casein diet with oleuropein supplementation than in those fed the 40% casein diet without oleuropein supplementation. In Experiment 2, the effects of oleuropein aglycone (a major phenolic compound in extra virgin olive oil and the absorbed form of oleuropein ingested in the gastrointestinal tracts) on the secretion of luteinizing hormone (LH) from the pituitary gland, which regulates testosterone production in the testis, were investigated in anesthetized rats. Plasma LH level increased dose dependently after the administration of oleuropein aglycone (P<.001, r= 0.691). These findings suggest that dietary supplementation with 0.1 g/100 g oleuropein alters the levels of hormones associated with protein anabolism by increasing urinary noradrenaline and testicular testosterone levels and decreasing plasma corticosterone level in rats fed a high-protein diet.     

Dactylospora mangrovei sp. nov. (Discomycetes, ascomycota) from mangrove wood

A new speciesDactylospora mangrovei is described and illustrated and compared withD. haliotrepha andD. canariensis, all marine inhabiting species.Dactylospora haliotrepha andD. mangrovei are tropical/sub-tropical mangrove species collected in Hong Kong, Malaysia, Taiwan and Thailand, whileD. canariensis was described from temperate waters. A map of the geographical distribution of the three marineDactylospora species is presented.     

Soy protein,casein, and zein regulate histidase gene expression by modulating serum glucagon

Glucagon has been postulated as an important physiological regulator of histidase (Hal) gene expression; however, it has not been demonstrated whether serum glucagon concentration is associated with the type and amount of protein ingested. The purpose of the present work was to study the association between hepatic Hal activity and mRNA concentration in rats fed 18 or 50% casein, isolated soy protein, or zein diets in a restricted schedule of 6 h for 10 days, and plasma glucagon and insulin concentrations. On day 10, five rats of each group were killed at 0900 (fasting), and then five rats were killed after being given the experimental diet for 1 h (1000). Rats fed 50% casein or soy diets showed higher Hal activity than the other groups studied. Rats fed 50% zein diets had higher Hal activity than rats fed 18% casein, soy, or zein diets, but lower activity than rats fed 50% casein or soy diets. Hal mRNA concentration followed a similar pattern. Hal activity showed a significant association with serum concentrations of glucagon. Serum glucagon concentration was significantly correlated with protein intake. Thus the type and amount of protein consumed affect Hal activity and expression through changes in serum glucagon concentrations.