Quality of eggs from Lohmann Brown Classic laying hens fed black soldier fly meal as substitute for soya bean

Soya bean is the main protein source in poultry feed but rising prices make an alternative protein source necessary. Insects, such as the black soldier fly (Hermetia illucens), may be an attractive solution for hens, although little information is available on their effect on egg quality. The present study aims to fill this gap by testing the effect of 100% replacement of soya bean with H. illucens larva meal in the diet of Lohmann Brown Classic laying hens for 21 weeks. At the end of the trial, the eggs were characterized for parameters such as weight, colour, proximate composition of albumen and yolk, and content of carotenoids, tocopherols and cholesterol. The fatty acid profile of yolks was also determined. Hens fed the insect-based diet produced eggs (HIM group) with a higher proportion of yolk than the group fed the soya bean-based diet (SBM group). HIM was associated with redder yolks (red index 5.63 v. 1.36) than SBM. HIM yolks were richer in γ-tocopherol (4.0 against 2.4 mg/kg), lutein (8.6 against 4.9 mg/kg), β-carotene (0.33 against 0.19 mg/kg) and total carotenoids (15 against 10.5 mg/kg) than SBM yolks. The fatty acid composition of HIM yolks was almost identical to that of SBM yolks. Finally, HIM yolks contained 11% less cholesterol than SBM yolks. These results suggest that H. illucens larva meal is a suitable total substitute for soya bean meal in the diet of Lohmann Brown Classic laying hens. A sustainable alternative to the plant protein source therefore seems feasible.     

Effects of fermented oyster mushroom ( Pleurotus ostreats) by-product supplementation on growth performance, blood parameters and meat quality in finishing Berkshire pigs

The objective of the present study was to investigate the effects of fermented oyster mushroom (Pleurotus ostreats) by-production (FOMP) supplementation on the growth performance, blood parameters, carcass traits and meat quality in finishing Berkshire pigs. FOMP was made by mixing oyster mushroom by-production with rice bran and barley bran and this mixture was fermented for 60 days. The experimental diets were 0, 3, 5 and 7% of FOMP added to C, T1, T2 and T3 in the basis diet for 7 weeks. Average daily gain (kg/day) was higher in C and T1 than in T2 and T3 ( P < 0.05). Average daily feed intake (kg/day) and feed conversion increased by the addition of FOMP ( P < 0.05). Total cholesterol and high-density lipoprotein cholesterol were higher in T3 than other treatments ( P < 0.05). Carcass weight (kg) was higher in C and T1 than in T2 and T3 ( P < 0.05). Dressing (%) was higher in C than in T3 ( P < 0.05). Crude protein was lower in T3 than in other treatments ( P < 0.05). Crude fat was higher in T2 and T3 than in C ( P < 0.05). pH₂₄ was higher in C than in other treatments ( P < 0.05). Cooking loss (%) was higher in T1 than T2 ( P < 0.05). Water-holding capacity (%) was higher in C than in T1 ( P < 0.05). In meat colour, CIE a* was lower by the addition of FOMP ( P < 0.05). CIE b* was higher in C than in other treatments ( P < 0.05). In backfat colour, CIE L* was lower in T3 than other treatments ( P < 0.05). CIE b* was lower by addition of FOMP ( P < 0.05). Palmitoleic and oleic acid were higher in T3 than in other treatments ( P < 0.05). Linoleic and arachidonic acids were higher in T2 than in other treatments ( P < 0.05). The results indicate that 3% of FOMP affected the growth performance, carcass traits, meat quality and fatty acid in contrast to addition of 5% of FOMP for Berkshire pigs during the finishing period.     

Effect of dietary supplementation of Ligustrum lucidum on performance,egg quality and blood biochemical parameters of Hy-Line Brown hens during the late laying period

The fruit of Ligustrum lucidum (FLL, Nuzhenzi in Chinese) is an important traditional medicine, and have attracted significant research attention because of their various biological activities. However, there are few research reports available on the use of FLL as a feed additive in livestock nutrition, particularly in layers. This study was conducted to determine the effects of supplementation of the diet of laying hens with FLL on laying performance, egg quality and blood metabolites. A total of 360 72-week-old hens were allocated to three dietary treatments (eight replications of 15 hens/treatment group) and were fed either a control diet or a diet supplemented with an inclusion level of 0.25% or 0.50% of FLL powder in the final feed, until 78 weeks of age. Hens were housed in a three-tier cage system. Feed and water were provided ad libitum. Blood samples and eggs were collected at the end of the experiment. The results showed that dietary supplementation with FLL did not affect egg weight, feed conversion ratio, eggshell thickness, albumen height, egg yolk color, eggshell breaking strength or egg shape index. However, FLL supplementation significantly decreased (P<0.001) mortality, cracked-egg rate and blood serum levels of cholesterol, low-density lipoprotein cholesterol, triglycerides and alanine aminotransferase, and increased (P<0.001) blood serum levels of high-density lipoprotein cholesterol. No differences in serum levels of total protein, albumin, glucose, calcium, aspartate aminotransferase or alkaline phosphatase were observed in hens fed FLL compared with the control group. It can be concluded that FLL, at a supplementation level of 0.25% final feed, can be used as an effective feed additive to improve the performance of laying hens during the late laying period.     

Interactions of flavin containing monooxygenase 3 (FMO3) genotype and feeding of choline and rapeseed cake on the trimethylamine (TMA) content in egg yolks of laying hens

To evaluate the relationship between the FMO3 genotype and dietary trimethylamine (TMA) precursors on TMA-N content in eggs, laying hens were fed with choline (500 and 4000 mg/kg diet) and, in a later trial, with rapeseed cake (0, 100 and 300 g/kg diet). Hens were genotyped for non-synonymous A/T polymorphism at position nt1034 of the cDNA of the FMO3 gene, which is associated with elevated TMA-levels in egg yolks. Genotypes were equally distributed among the experimental diets. TMA was extracted from yolks and measured as TMA-N by a colorimetric method. As expected, hens of the TT-genotype had high amounts of TMA-N in egg yolks when fed either 4000 mg choline/kg diet or the rapeseed cake diet, while hens of the AA-genotype showed low levels of TMA-N. Heterozygous AT-hens had a slightly increased level of TMA-N when fed with 4000 mg choline/kg diet but not with the rapeseed cake feed. Rapeseed cake can be fed to AA and AT hens without leading to fishy tainted eggs.     

The impacts of fermented feed on laying performance,egg quality,immune function,intestinal morphology and microbiota of laying hens in the late laying cycle

Fermented feed has the potential to improve poultry gastrointestinal microecological environment, health condition and production performance. Thus, the present study was undertaken to explore the effects of fermented feed on the laying performance, egg quality, immune function, intestinal morphology and microbiota of laying hens in the late laying cycle. A total of 360 healthy Hy-Line Brown laying hens aged 80 weeks were used to conduct a 56-day study. All hens were randomly separated into two treatment groups, with five replicates of 36 hens each as follows: basal diet containing 0.0% fermented feed (CON) and 20% fermented feed (FF). Subsequent analyses revealed that fermented feed supplementation was associated with significant increases in laying rates together with reduced broken egg rates and feed conversion ratio for hens in FF group (P < 0.05). There were additionally significant increases in both albumen height and Haugh unit values in hens following fermented feed supplementation (P < 0.05). Fermented feed was also associated with increases in duodenal, jejunal and ileac villus height (P < 0.05). Laying hens fed fermented feed had higher immune globulin (Ig)A, IgG, IgM levels (P < 0.01,) and higher interleukin 2, interleukin 6, tumour necrosis factor α and interferon γ (P < 0.05) concentrations than CON. Analysis of the microbiota in these laying hens revealed the alpha diversity was not significantly affected by fermented feed supplementation. Firmicutes abundance was reduced in caecal samples from FF hens relative to those from CON hens (30.61 vs 35.12%, P < 0.05). At the genus level, fermented feed was associated with improvements in relative Lactobacillus, Megasphaera and Peptococcus abundance and decreased Campylobacter abundance in laying hens. These results suggest that fermented feed supplementation may be beneficial to the laying performance, egg quality, immunological function, intestinal villus growth and caecal microecological environment of laying hens at the end of the laying cycle.     

The enzymatic conversion of cis-(14C)phytofluene, trans-(14C)phytofluene, and trans-zeta-(14C)carotene to more unsaturated acyclic, monocyclic, and dicyclic carotenes by a cell-free preparation of red tomato fruits

This paper reports the conversion of cis-[¹⁴C]phytofluene to trans-[¹⁴C|phytofluene and the conversion of the latter compound to trans-ζ-[¹⁴C]carotene by a soluble enzyme system obtained from the plastids of red tomato fruits. Each of these radioactive compounds was also converted to labeled neurosporene, lycopenc, α-carotene, and β-carotene by the same enzyme system. The incorporation of each substrate into more unsaturated carotenes was carried out under nitrogen at pH 7.5–8.2 (borate buffer), at 25 °C in the dark.Proof of the formation of the above carotenes from each of the three radioactive substrates was demonstrated by cochromatography with authentic nonradioactive carotenes on an alumina chromatographic column. A close correspondence between radioactivity and light absorbance for each carotene was observed. Confirmation of these conversions was achieved by cochromatography with authentic samples on thinlayer plates. Final proof for the formation of the acyclic and cyclic carotenes from the above radioactive substrates was obtained by gas-liquid chromatography of the hydrogenated products. Coincidence between mass and radioactivity was observed.Maximum conversion of cis- and trans-phytofluenes to more unsaturated carotenes by the red tomato fruit enzyme system appears to be dependent upon the presence of NADP⁺, FAD, and Tween 80. The formation of the carotenes is also increased in the presence of Mg²⁺ or Mn²⁺ ions.     

Effect of the ratio of (+) and (−) mating type of Blakeslea trispora on carotene production from cheese whey in submerged fermentation

The effect of the ratio of (+) and (?) mating type of Blakeslea trispora on carotene production from deproteinized hydrolysed whey in shake flask culture was investigated. Also, the inoculum ratio of the two mating types on the morphology of the microorganism and the relationship between morphological changes of the fungus and product formation were studied. The concentration of carotenes was significantly affected by the ratio of (+) and (?) mating type of B. trispora. A ratio of 1:10 up to 1:100 of (+) and (?) type was found to achieve the highest carotene yields. The optimum ratio of the (+) and (?) mating types for the maximum pigment production (175.0 mg/g dry biomass at 8 days of fermentation) was found to be 1:10. The carotene content in B. trispora consisted of β-carotene, γ-carotene, and lycopene. At the maximum concentration of carotenes the proportion of β-carotene, γ-carotene, and lycopene (as percent of total carotenes) was 80, 12, and 8%, respectively. B.trispora growing in submerged fermentation is able to develop complex morphologies which have been classified into three major groups: freely dispersed hyphae, clumps and pellets. These parameters are strongly influence the production of carotenes.     

Melamine in eggs,plasma and tissues of hens fed contaminated diets

A study was conducted to evaluate the excretion pattern of melamine from feed into eggs, plasma, kidney, liver and muscle of laying hens. In particular, 90 laying hens were randomly allocated to three dietary treatments and fed diets contaminated with melamine at a level of 2.5, 25 and 250 mg of melamine/kg of diet for T1, T2 and T3 groups, respectively. The diets were offered in six replicate boxes (five hens each) for 13 days. Eggs were collected from each group for melamine quantification on days 0, 1, 3, 6, 9 and 13. At the end of the experimental period, one hen per box was randomly selected and slaughtered to collect plasma, liver, kidney and muscle samples. During the experiment, feeding diets with increasing levels of melamine had no effect (P > 0.05) on weight gain, feed intake, egg production, egg weight and mortality of laying hens. The melamine in eggs increased from day 1 after melamine ingestion and reached a plateau between days 6 and 13 of melamine ingestion. At steady-state condition, the melamine egg concentrations increased (P < 0.01) with treatments, being 0.026, 0.352 and 4.631 mg/kg for T1, T2 and T3, respectively. Similarly, the carryover of melamine from feed to egg increased (P < 0.05) with the levels of melamine in the diets, varying from 0.50 to 0.70 and 0.84 for T1, T2 and T3, respectively. The melamine was detected in plasma of all tested groups, increasing (P < 0.01) with levels of melamine in the diets (0.030, 0.266 and 4.102 mg/l in T1, T2 and T3, respectively). Melamine was not detected in kidney, liver and muscle of hens fed T1. Except for kidney sampled in the T3, no melamine concentration higher than 2.5 mg/kg, representing the maximum allowable limit set by the US Food and Drug Administration and European Union for food and feeds, was measured. The melamine resulted higher in plasma and kidneys than in the liver and muscle both in T2 and T3. The results confirmed the presence of an excretion pattern of melamine from feed to eggs and tissues in laying hens.     

The isolation, purification, and characterization of cis-zeta-carotene and the demonstration of its conversion to acyclic, monocyclic and dicyclic carotenes by a soluble enzyme system obtained from the plastids of tangerine tomato fruits

Cis-ζ-carotene was isolated, purified in several chromatographic systems, and then identified as an intermediate in the biosynthesis of poly-cis-carotenes. The structure of cis-ζ-carotene was tentatively established from its visible light-absorption spectrum, and by a comparison of the infrared spectrum with that of trans-ζ-carotene. Confirmation of the identity of this compound was obtained by high resolution mass spectroscopy. The presence of the cis-configuration was indicated by a bathochromic shift of 6–10 nm in the visible spectrum when the compound was subjected to iodinecatalyzed photoisomerization. The infrared spectrum also showed characteristic peaks for the cis-configuration. Proof of the conversion of cis-ζ-[¹⁴C]carotene to trans-ζ-carotene, proneurosporene, prolycopene, neurosporene, lycopene, β- and γ carotenes was obtained on incubation with soluble enzyme systems obtained from plastids of fruits of two different tangerine varieties of tomato. Proof for the formation of each of the carotenes was provided by column and thin-layer chromatography A close correspondence of radioactivity and optical density was observed for each carotene. Additional proof was obtained by gas-liquid chromatography of each hydrogenated carotene. A coincidence of mass and radioactivity was observed for each carotene.     

β-Carotene biosynthesis by extracts of the C115 mutant of Phycomyces blakesleeanus

A cell extract of the yellow C115 car-42 mad-107(?) mutant of Phycomyces blakesleeanus, capable of converting MVA-[2-¹⁴C] into isoprenoids, was used to investigate the formation of β-carotene. The incorporation of radioactivity into β-carotene was reduced by the addition of unlabelled carotenes, solubilised using detergent, to the incubation mixtures. On reisolation of these carotenes after anaerobic incubations, they were found to carry radioactivity. The relative efficiencies of these carotenes as trapping agents are discussed in relation to the pathways of carotene cyclisation and to the apparent operation of a system for the negative feedback control of carotene biosynthesis.     

The enzymatic conversion of cis-(14C)phytofluene, trans-(14C)phytofluene, and trans-zeta-(14C)carotene to poly-cis-acyclic carotenes by a cell-free preparation of tangerine tomato fruit plastids

The conversion of cis-[¹⁴C]phytofluene to trans-[¹⁴C]phytofluene and the conversion of the latter to trans-ζ-[¹⁴C]carotene by a soluble enzyme system obtained from plastids of tangerine tomato fruits is reported. Each of these compounds is also converted to cis-ζ-carotene, proneurosporene, prolycopene, neurosporene, lycopene, and γ- and β-carotenes. [¹⁴C]Prolycopene was also incubated with the above enzyme system. No conversion of this compound to trans-lycopene or cyclic carotenes was observed. Proof for the formation of the above carotenes from each of the substrates mentioned above was obtained by cochromatography with authentic samples on an alumina column. A close correspondence between radioactivity and light absorbance of each carotene was observed. Further proof for the formation of acyclic and cyclic carotenes from the above radioactive substrates was obtained by gas-liquid chromatography of the hydrogenated products. Coincidence between mass and radioactivity was observed in each case.     

The role of oxidative stress on carotene production by Blakeslea trispora in submerged fermentation

In aerobic metabolism, reactive oxygen species (ROS) are formed during the fermentation that can cause oxidative stress in microorganisms. Microbial cells possess both enzymatic and non-enzymatic defensive systems that may protect cells from oxidative damage. The antioxidant enzymes superoxide dismutase and catalase are the two key defensive enzymes to oxidative stress. The factors that induce oxidative stress in microorganisms include butylated hydroxytoluene (BHT), hydrogen peroxide, metal ions, dissolved oxygen tension, elevated temperature, menadione, junglone, paraquat, liquid paraffin, introduction to bioreactors of shake flask inocula and synthetic medium sterilized at initial pH 11.0. Carotenes are highly unsaturated isoprene derivatives. They are used as antioxidants and as coloring agents for food products. In fungi, carotenes are derived via the mevalonate biosynthesis pathway. The key genes in carotene biosynthesis are hmgR, ipi, isoA, carG, carRA and carB. Among microorganisms, Βlakeslea trispora is the main microorganism used for the production of carotenes on the industrial scale. Currently, the synthetic medium is considered the superior substrate for the production of carotenes in a pilot plant scale. The fermentation systems used for the production of carotenes include shake flasks, stirred tank fermentor, bubble column reactor and flat panel photobioreactor. This review summarizes the oxidative stresses in microorganisms and it is focused on the current status of carotene production by B. trispora including oxidative stress induced by BHT, enhanced dissolved oxygen levels, iron ions, liquid paraffin and synthetic medium sterilized at an initial pH 11.0. The oxidative stress induced by the above factors increases significantly the production of carotenes. However, to further reduce the cost of carotene production, new biotechnological methods with higher productivity still need to be explored.     

Natural variation of carotenoids in the eggs and gonads of the echinoid genus, Strongylocentrotus: implications for their role in ultraviolet radiation photoprotection

We examined variability in carotenoid concentration in the gonads and eggs of four sea urchin species (Strongylocentrotus purpuratus, Strongylocentrotus franciscanus, Strongylocentrotus pallidus and Strongylocentrotus droebachiensis) to explore the possible role of carotenes as photoprotectants. Carotene concentrations were measured in gonads and gametes of each species, while in eggs the ultraviolet radiation (UV-R) sensitivity and self-shading capacity by carotenes were calculated. Mean concentrations of carotenes in gonads ranged from 0.13±0.017 mg g⁻¹ dw (S. purpuratus), 0.14±0.019 mg g⁻¹ dw (S. franciscanus), 0.29±0.079 mg g⁻¹ dw (S. pallidus) to 0.36±0.06 mg g⁻¹ dw (S. droebachiensis). In eggs, concentrations ranged from 0.026±0.003 to 0.09±0.034 mg g⁻¹ dw. UV-R sensitivity in eggs was quantified by measuring UV-R induced first-cleavage delay. Intra-specifically, cleavage delay varied significantly between individuals, and could be correlated with carotene concentration. Interspecific differences in cleavage delay and carotene concentrations were not correlated. Using the observed concentration of β, β-echinenone (which makes up between 82.4% and 94.9% of the total carotene concentration in the eggs) and a molar extinction coefficient of ε=13.7×10³ mol⁻¹ cm⁻¹ at 334 nm, we calculated self-shading efficiency in the eggs. Self-shading capacity (J₃₃₄) indicated that the eggs could only screen from 4.6% (J₃₃₄=0.046) down to 1.5% (J₃₃₄=0.015) of UV-R at 334 nm. While not sunscreens, we suggest that carotenes can photoprotective in echinoid eggs, probably by mitigating the effects of reactive oxygen species.     

Stimulation of the biosynthesis of carotenes by oxidative stress in Blakeslea trispora induced by elevated dissolved oxygen levels in the culture medium

The adaptive response of the fungus Blakeslea trispora to the oxidative stress induced by elevated dissolved oxygen concentrations during carotene production was investigated by measuring the specific activities of catalase (CAT) and superoxide dismutase (SOD) and the micromorphology of the fungus using a computerized image analysis system. Changes in the ratio of the volume of air (V_a) over the medium and the volume of medium (V_m) in the flask caused changes of the morphology of microorganism from clumps to pellets and increases in the specific activities of CAT and SOD. The oxidative stress in B. trispora resulted in a significant increase in carotene production, and a maximum proportion of β-carotene (60%), γ-carotene (50%), and lycopene (10%) (as percentages of total carotenes) was observed at a ratio V_a/V_m of 15.7, 4.0 and 1.5, respectively. The highest concentration of carotenes (115.0 mg/g dry biomass) was obtained in V_a/V_m ratio of 9.0.     

Effect of dietary Ptecticus tenebrifer powder on the fatty acid profile of egg yolk in laying hens

The poultry industry needs alternative feeds with no adverse effects on animal performance and egg quality. Insects represent effective animal feeds. However, Ptecticus tenebrifer (PT) has rarely been studied as an effective feed in terms of fatty acid contents. Therefore, we investigated the effect of dietary PT powder on the fatty acid profiles of egg yolks in laying hens. A total of 180 Hy-Line brown hens were divided equally into three groups (n = 60), with three replicates: the control group and two groups fed 2% and 4% PT powder, respectively. After 2 weeks, minor effects (P < 0.05) on oleic acid, dihomo-gamma-linolenic acid, and total monounsaturated fatty acids were observed among the groups. After 4 weeks, docosahexaenoic acid, docosapentaenoic acid, myristic acid, oleic acid, and total polyunsaturated fatty acids were slightly altered in the treatment groups (P < 0.05). For other fatty acids, the 2% and 4% PT groups had fatty acid concentrations similar to those of the controls. No differences in total n-6 and total n-3 were observed among the groups. In conclusion, using 2% and 4% PT powders did not notably change the fatty acid profiles of egg yolks and could replace some layer-hen feed without negative effects.     

Does Dietary vitamin E or C Decrease Egg Yolk Cholesterol?

An experiment was conducted to determine the effect of dietary vitamin E and C on serum metabolites, yolk cholesterol, egg quality, and performance of layer hens. One hundred sixty-eight commercial Hy-Line W-36 layer hens were randomly divided into seven groups and six replicates with four hens in each. Dietary treatments were introduced after the pre-experimental period (10 days) to adjust egg production. Treatments were levels of vitamin E or C (100, 200, and 400 mg/kg diet) supplementation to the basal diet for 4 weeks, whereas the control group received no supplementation. Egg production, egg weight, and feed consumption were recorded during the study. Shell thickness, Haugh unit score, yolk color, yolk weight, yolk cholesterol, and blood parameters were measured at the end of experiment. There was no significant effect of dietary vitamin E or C on hen performance. Egg yolk cholesterol concentrations decreased linearly by antioxidant vitamin supplementation (P?<?0.01). Egg yolk cholesterol reduction did not have any negative effect on egg production rate. Antioxidants, especially vitamin C, increased serum glucose concentration (P?<?0.05). Serum total cholesterol content did not change by vitamin supplementation but cholesterol in high-density lipoprotein (HDL-C) decreased and cholesterol in low-density lipoprotein (LDL-C) increased (P?<?0.05), as dietary vitamin E or C supplementation increased in diets. These results are in conflict with the previous hypothesis that antioxidants have a role in LDL-C removal from the blood or increasing HDL-C. Vitamin E was more effective than vitamin C in this case and if these results are confirmed by further studies, they may result to revision in researchers’ point of view about antioxidant especially in human medicine.     

Using a feed-grade zinc propionate to achieve molt induction in laying hens and retain postmolt egg production and quality

A commercial-feed-grade form of zinc propionate was examined as a potential feed amendment at a concentration of 1% zinc to induce molt in 90-wk-old hens. Dietary treatments consisted, of 4 treatment groups of 28 birds each randomly assigned to either (1) molted conventionally by feed withdrawal, (2) 1% zinc as Zn acetate, (3) 1% zinc as Zn propionate, or (4) nonmolted control for 9 d. Ovary weights of hens fed Zn acetate or Zn propionate were not significantly different from each other, but hens fed Zn acetate or Zn propionate were significantly (p<0.05) lighter than the ovary weight of nonmolted control hens. Zinc concentrations in the kidney and liver were significantly (p<0.05) increased in both Zn acetate- and Zn propionate-molted hens when compared to either nonmolted control-fed hens or feed-withdrawal molted hens. Over the entire 3-mo postmolt period, there were no significant differences in interior or exterior egg qualities among the four treatments Egg production of hens fed Zn acetate was significantly lower than feed-withdrawal hens, Zn propionate-fed hens, or nonmolted control hens (p<0.05). The data of the current study demonstrated that feeding a feed grade of Zn propionate (1% Zn)-supplemented diet can induce molt and retain postmolt egg quality and production comparable to hens molted by feed withdrawal.     

Investigations into the effects of processing on the retention of the carotenoid fractions of Leucaena leucocephala during storage,and the effects of processing on mimosine concentration

Leucaena leucocephala leaves were subjected to several drying treatments followed by pelleting and/or inclusion of ethoxyquin as antioxidant and analysed monthly to determine rates of loss of carotenes and xanthophylls. Rapid sun drying yielded leucaena leaf meal with carotene and xanthophyll concentrations of 446 and 865 mg/kg, respectively, but considerable losses occurred during drying at 60 and 145°C, and as a result of blanching in hot water or ferrous sulphate solution. Loss rates of carotenes during storage were in the range 19–40 mg/kg per month, and of xanthophylls, 29–53 mg/kg per month. Carotenoids in sun-dried leaf meal were most stable during storage. Pelleting or inclusion of ethoxyquin did not reduce carotenoid loss rates below those of unpelleted leaf meal.The mimosine content of untreated sun-dried leucaena leaf meal was 3.2%, but the concentration decreased to 2.5 and 1.8% on drying at 60 and 145°C, respectively. The concentrations of mimosine in sun-dried leafmeals of water-blanched and ferrous sulphate-blanched leucaena were 2.0 and 2.1%, respectively. Treatments causing maximum losses of mimosine were those which brought about maximum losses of carotenoids.     

Metabolism of cholesterol in the tissues and blood of the chick embryo

Three artificially inseminated laying White Leghorn hens were given 35-50 micro c of cholesterol-4-(14)C intravenously. Their subsequently produced eggs contained cholesterol-(14)C-labeled yolks. Some of the fertilized eggs were analyzed for cholesterol content and radioactivity. Other eggs were incubated until hatching. The specific activity of the cholesterol contained in the serum and tissues of newly hatched chicks was determined and compared with that of yolk sac, which was taken as representative of egg yolk cholesterol before its metabolic transfer into the chick embryo. The specific activities of cholesterol in intestine, liver, serum, heart, and skeletal muscle and the whole chick were 95-98% of that in yolk sac, but that of brain cholesterol was only 11% of this value. These results indicate that whereas most of the cholesterol in the chick originated from the egg yolk, cholesterol biosynthesis was active in the brain and provided about 90% of its cholestero content. Newly hatched chicks were found to be hyperlipemic compared with older chicks and had fatty livers with a high cholesterol content. Desmosterol was found in 9- and 15-day old chick embryos but not in the newly hatched chicks, in which the only sterol was cholesterol.     

Ultraviolet light sensitivity of carotene-and cytochrome-c-accumulating strains of the smut fungusUstilago violacea

White, pink, orange, and yellow strains ofUstilago violacea containing high and low levels of cytochrome c and various carotenes were exposed to ultraviolet light. The survival curves for all strains were of exponential decay form, but the carotene-accumulating strains were generally more resistant to UV than those strains with no carotenes at all. The UV exposure time leading to 90% loss in viability, LD₉₀, was quantitatively related to the carotene content in the form of a power function, where LD₉₀ increased as the fifth root of the total carotene content per cell. We also determined that the ratio of total carotenes to total cytochrome c per cell was quantitatively related to the rate of viability loss during UV exposure.