Eggshell Ca Research Articles

Implementation of eggshell extracted calcium acetate in biocementation via soybean urease

Biocementaion, a promising soil improvement technique, mostly utilizes calcium chloride (CaCl2) as its calcium source. However, using CaCl2 poses a critical environmental problem of producing ammonium chloride as a by-product. Alternatively, when calcium acetate (Ca(CH3COO)2) is used as the calcium source, the production of ammonium chloride during biocementation can be reduced. Ca(CH3COO)2 can be easily derived from waste eggshells. Nevertheless, such research has been conducted only with Microbial Induced Carbonate Precipitation (MICP) to date. This paper aims to study the implementation of eggshell Ca(CH3COO)2 in the more effective biocementation approach: Enzyme Induced Carbonate Precipitation (EICP) via soybean urease. For this study, EICP solutions of varying molar ratios and concentrations were studied for their precipitation efficiency and reinforcement effect on poorly graded sandy soil. The findings showed that the EICP solution composition of 1:1 molar ratio between Ca(CH3COO)2 and urea, added with 50 g/L soybean solution reached the maximum precipitation efficiency. It also produced the highest 7-day UCS of 371 kPa at a calcium carbonate content of 0.40%. Furthermore, the eggshell-soybean EICP produced 8.5% higher UCS than CaCl2-soybean EICP despite having similar calcium carbonate content.

Assessing Bone Health Status and Eggshell Quality of Laying Hens at the End of a Production Cycle in Response to Inclusion of a Hybrid Rye to a Wheat-Corn Diet.

The objective of this study was to evaluate whether there are negative effects of the partial replacement of white corn with rye along with xylanase supplementation on overall bone quality, eggshell mineralization, and mechanical strength in laying hens. From the 26th week of life, ISA Brown laying hens were fed either a wheat-corn diet or a diet containing 25% rye, with or without xylanase. The experimental period lasted for 25 weeks, until birds reached their 50th week of age, after which bone and eggshell quality indices were assessed. Eggshell thickness and eggshell Ca content of eggs from rye-fed hens were improved by xylanase supplementation. No differences in the mechanical properties of the eggshells were observed between treatments, except for the diet-dependent changes in egg deformation. Rye inclusion had no effect on the mechanical properties of bone. Xylanase supplementation, irrespective of the diet, had a positive effect on bone strength and increased tibia Ca content, as well as the content of several microelements. Hence, hybrid rye combined with wheat can replace 25% of corn in layer diets without compromising shell quality or bone mineral content. Xylanase supplementation in these diets is recommended since its inclusion improves both bone strength and quality.

Indirect Measurement of the Amounts of Casein Phosphopeptides Added to Dangmyon (Starch Vermicelli or Glass Noodle) as a Calcium-Absorption Enhancer

Industrial-scale Ca-fortified Dangmyon was manufactured with added casein phosphopeptides (CPP) to increase Ca adsorption in the intestine. Very low P content in eggshell Ca (egg Ca) and Dangmyon could make it possible to indirectly measure CPP content in Dangmyon. Partitions of the whole P present in Dangmyon were made into sweet potato, egg Ca, and CPP. The CPP content was obtained by multiplying CPP per P by the amount of P partitioned into CPP. It was found that 88.46% of CPP was obtained. However, when milk Ca, which was much higher in P, was added to fortify Dangmyon with CPP, it was found that the CPP content was either under- or over-estimated. Care must be taken when a much higher content of P as a Ca source is selected using this method. It was discovered that the added Ca and CPP were retained after cooking at boiling temperature; therefore, Dangmyon could be an excellent Ca and CPP carrier for humans.

Minerals Composition and Characterization of the Hatchery Eggshell Waste Treated with Different Preparation Methods

The preparation process needs to be carried out in the treatment of hatchery waste. The objective of the study was to determine the mineral content and characterization of the eggshell waste after preparation. An experiment was arranged on a completely randomized design (CRD). Preparation treatment consisted of; T1: shell waste watered with distilled water; T2: shell waste soaked in distilled water for 12 h; T3: shell waste soaked in 0.5% NaOH for 720 min; and T4: shell waste boiled at 80°C for 15 min and soaked in 5% H3PO4 for 15 min. The data were analyzed using Analysis of Variance (ANOVA), and if any differences, a Duncan Multiple Range Test (DMRT) post hoc tests were carried out. Composition of proximate and mineral were detected by using X-Ray Fluorescence (XRF). Results indicated that proximate analysis of the dry matter content were T4: 98.98±0.01; T3: 98.58±0.04; T2: 98.75±0.08; and T1: 98.72±0.03. The calcium content of the treatment from the highest was T3: 24.22±0.31; T1: 22.80±0.57; T2: 22.77±0.71; and T4: 21.55±0.46. It may be inferred that the treatment technique had no major impact on the eggshells physical characteristics. However, boiling treatment at 80°C for 15 min and soaking in 5% H3PO4 reduced the eggshell waste's Mg, Si, and Ca content. The addition of 0.5% NaOH immersion did not degrade Mg, Ca, and Si of eggshell waste.

Effects of dietary Bacillus subtilis supplementation and calcium levels on performance and eggshell quality of laying hens in the late phase of production

The objective of this study was to evaluate the effects of dietary Bacillus subtilis supplementation and calcium (Ca) levels on performance, eggshell quality, intestinal morphology, and relative calbindin-D28k (CALB1) mRNA level of laying hens in the late phase of production. An experiment employing a 2 × 3 factorial arrangement of 3 levels of Ca (3.5, 4.0, and 4.5%) and the absence or presence of B. subtilis was carried out with a total of 576 Hy-Line Brown laying hens aged 72 to 79 wk. Every group had 8 replicates of 12 birds each. The results showed that 4.0 and 4.5% Ca levels improved (P < 0.05) apparent retention and serum Ca content of aged laying hens. Compared with the 3.5% Ca level, the 4.0% Ca level in diets increased (P < 0.05) thickness, eggshell weight, shell ratio, and eggshell Ca content of aged laying hens. Moreover, breaking strength, thickness, eggshell weight, shell ratio, eggshell Ca content, apparent retention of Ca in g/day, apparent retention of Ca in percent, villus height, villus height/crypt depth, serum Ca level, and relative CALB1 mRNA level of aged laying hens were all increased (P < 0.05) by B. subtilis supplementation in diets. The supplemental B. subtilis decreased feed conversion ratio (P = 0.001) significantly. In addition, there was an interaction effect between increased Ca levels from 3.5 to 4.5% and B. subtilis supplementation on crypt depth in the duodenum (P < 0.05). In conclusion, we found that both the increase in dietary Ca level from 3.5 to 4.5% and B. subtilis supplementation could enhance intestinal Ca absorption and improve eggshell quality of laying hens in the late phase of production (72–79 wk of age). Dietary supplementation of B. subtilis accompanying the 4.0% Ca level was appropriate in enhancement of eggshell quality.

Productive performance, bone characteristics, and intestinal morphology of laying hens fed diets formulated with L-glutamic acid

This study was conducted to evaluate the effects of L-glutamic acid (Glu) on the productive performance, eggshell quality, bone characteristics, and morphologic parameters of laying hens. Two hundred and forty 53-wk-old Hy-Line W36 laying hens were randomly allocated into 5 treatments, 6 replicates, and 8 birds each. The experimental period was 16 wk. The treatments consisted of a Basal diet (Glu calculated at 2.68%) and 4 L-glutamic acid inclusion levels: 2.88, 3.08, 3.28, and 3.48%. Glu inclusion levels in the hens diet had an increasing linear effect (P < 0.05) on eggshell Ca concentration and bone (tibiotarsus) strength index, and a decreasing linear effect (P < 0.05) on the Seedor index. Serum biochemistry results for total and ionic Ca, inorganic P, and alkaline phosphatase were affected by blood collection time (04:00 Pm, 03:00 Am, and 12:00 Pm) but not by the treatments. Jejunum morphometric variables were not influenced by the treatments except for crypt depth, which demonstrated a quadratic effect (P < 0.05). However, proliferating cell nuclear antigen qualitative immunohistochemical analysis of the jejunum showed more positive nuclei in the villus with the addition of Glu. Both with the basal diet and the lowest Glu inclusion treatment, positive nuclei were observed in the crypts and at the base of the villus, while in the treatments with higher Glu levels, positive cells were common all along the villus mucosa, including its extremity. In conclusion, diets formulated with increased levels of Glu had beneficial effects on eggshell Ca concentration, tibiotarsus structure, and proliferative activity of the jejunum of 69-wk-old laying hens. Therefore, dietary L-glutamic acid can be considered an alternative additive to improve bone characteristics in the productive phase of laying hens.

Effects of Dietary Different Levels of Nano, Organic and Inorganic Zinc Sources on Performance, Eggshell Quality, Bone Mechanical Parameters and Mineral Contents of the Tibia, Liver, Serum and Excreta in Laying Hens.

This study was conducted to determine the effect of dietary zinc (Zn) sources and their levels on the performance, egg quality, tissue mineral concentrations and bone mechanical traits of laying hens. Two hundred seventy, 44-week-old, Super Nick white laying hens were randomly distributed to 15 experimental groups in a 3 (sources of Zn) × 5 (levels of Zn) factorial arrangement, using groups of 18 birds with 6 replicates. The 15 experimental diets consisted of three sources of Zn (zinc-oxide as an inorganic form, zinc-proteinate as an organic form and nano zinc-oxide powder as a nano form) and five different levels of Zn (20, 40, 60, 80 and 100mg Zn/kg diet). The experiment lasted 12weeks. Dietary Zn sources, Zn levels and their interactions had no significant effect on the performance parameters between the treatment groups, nor a significant effect on eggshell weight and eggshell breaking strength. Eggshell thickness and eggshell Ca and P contents were significantly affected by the different dietary Zn sources. Dietary Zn sources, Zn levels and their interactions had no significant effect on tibia Ca and P contents, and also had no significant effect on tibia Zn content. Liver Zn content was significantly affected by the dietary Zn sources. Serum Zn content was not affected by the dietary treatments. Dietary Zn sources, Zn levels and their interactions had no significant effect on tibia weight, tibia stress and tibia breaking strength, as tibia mechanical parameters. Excreta Zn content decreased linearly when hens were fed nano Zn compared to organic and inorganic Zn, and excreta Zn content also linearly increased with increasing dietary Zn levels. In conclusion, 20mg/kg of supplemental Zn is optimal for sustaining performance, good eggshell quality and bone status, while also reducing Zn excretion and soil pollution, with organic (Zn-proteinate) and nano (ZnO) Zn as the preferred forms in laying hen diets.

The Effect of Different Dietary Calcium Levels on Calcium Metabolism in Laying Hens

Abstract The experiment was carried out on four groups of Hy-Line var. Brown hens (n = 350 each) fed on diets which provided 2.49, 3.07, 3.78 and, respectively, 4.15 g% calcium (Ca) during the laying period. Consequently, mean daily Ca intake was 2.74, 3.38, 4.16 and 4.60 g/cap., respectively. Ca metabolism was assayed at 20 weeks of age (start of the laying), 36 weeks of age (the peak of the laying) and 68 weeks (the end of the laying period). A significant part of the ingested Ca was not absorbed. The percentage of Ca exonerated by feces was higher at the start of the laying and lowers in the peak of laying and at the end of the laying period. The amount of Ca exonerated as unabsorbed Ca increases as the amount of ingested Ca increases inside of each age level. It was found also an increase of the Ca content of the egg shell according the Ca intake statistically significant (P&lt;0.05) in 36- and 68-wk.-old hens, but not in 20-wk.-old hens. Blood plasma Ca showed significantly higher values in 3.38-, 4.16- and 4.60-g/day-Ca-intake groups when compared by 2.74-g/day-Ca-intake group (P &lt; 0.05) but the level of increasing showed lowest values in hen groups with highest intake of Ca. The daily eggshell Ca export strongly correlates with the amount of daily Ca intake in 36-wk.-old hens and 68-wk.-old hens: r = +0.72 and +0.84, respectively, but no correlation was found in 20-wk.-old hens (r = -0.74, negative correlation). The results showed a limited physiologic capacity of the Ca absorption in hen, the Ca surplus being lost. Dietary Ca level should be closely correlated with the laying level of hens.

Eggshell fractions containing different particle sized affect mineral absorption but not bone mineral retention in growing rats

The purpose of this study was to compare the calcium (Ca) bioavailability from eggshell fractions containing different particle size to purified CaCO3 in male growing rats. Mineral absorption, bone mineral concentration, and biomechanical properties were evaluated. Mean Ca absorption of rats fed with eggshell diets amounted to 56.2% of the ingested Ca, which is considered high. However, we observed lower Ca absorption in large-sized particle eggshell fraction (ES L) and small-sized particle eggshell fraction groups but similar Ca absorption in intermediate-sized particle eggshell fraction (ES M) compared with the CaCO3 group. Rats that received ES M and ES L had higher P and Mg absorption than the CaCO3 group. No changes were observed in the bone mineral deposition, weight or mechanical resistance. We conclude that eggshell Ca is well absorbed by the intestine and retained in bones of growing rats, being a low cost alternative to achieve adequate Ca ingestion.

Changes in Total Serum Calcium Level and Shell Calcium Content in Bank Myna, Acridotheres ginginianus (Latham) During Breeding Season

Bank Myna is a wild Passerine seasonal breeder of North India. Higher values of serum Ca had been documented in early breeding season which gradually reduced in late breeding season. Eggshell Ca content closely correspond the rise and fall in serum Ca level. Highest serum Ca level of 15.13+0.146mg/dl and eggshell Ca content of 357.25+0.310mg/g were recorded in first week of June.

Safety evaluation of daidzein in laying hens: Part II. Effects on calcium-related metabolism

Daidzein, an estrogen-like product, has become increasingly popular as a dietary supplement, particularly for postpeak-estrus animals seeking a safe natural alternative to play a role of estrogen. However, there is little available safety data of it for raisers and consumers. A subchronic laying hensafety study has been conducted to examine if the high-dose daidzein could affect calcium-related metabolism (eggshell quality and bone mineralization). Seven hundred and sixty-eight 56-week-old Hyline Brown were randomly assigned to 4 groups with 8 replicates of 24 birds each (192 laying hensper group) and 3weeks later fed diets supplemented with 0(control), 10, 50 and 100mg of daidzein/kg for 12week. Eggshell thickness, eggshell percentage, eggshell strength, eggshell Ca concentration was increased linearly with increasing dietary daidzein supplementation (P=0.001, P=0.007, P=0.002 and P=0.000, respectively). Serum Ca increased linearly with increasing dietarydaidzein supplementation (P=0.042), and serum P showed a significant quadratic response to dietarydaidzein supplementation (P=0.036). Bone ash and bone Ca were significantly influenced by dietarydaidzein supplementation (P<0.05). These findings indicate that daidzein hold no observed adverse effect on calcium metabolism, but also a safe and effective food additive for calcium metabolism in animals and humans.

Testing the structural–function hypothesis of eggshell maculation in the Great Tit: an experimental approach

The function of eggshell pigmentation has long been the subject of much speculation. Most of the proposed hypotheses have been based on a signalling function. According to the recently proposed structural–function hypothesis, birds use the pigment protoporphyrin to strengthen the eggshell in response to dietary calcium (Ca) shortage. The aim of the study reported here was to test this hypothesis in a Ca-provisioning experiment involving Great Tit Parus major populations breeding in Ca-poor coniferous and Ca-rich deciduous Estonian forests, respectively, over a 2-year period. Ca-supplemented birds laid eggs with ‘darker’ pigmentation than birds in the unsupplemented (control) group, which is at odds with the structural–function hypothesis. Although eggs were smaller and had thinner shells in Ca-poor habitat, eggshell maculation did not differ between the habitats. Eggshells were more pigmented in the year when they were also thinner, but their thickness was not related to their maculation within each year. Although the results of our study neither prove nor disprove the structural–function hypothesis, they do suggest that the relationship between eggshell pigmentation and Ca availability cannot be explained simply by this hypothesis.

Effect of Egg-Shell Ca on Preventing of Bone Loss after Ovariectomy

Effect of Egg-Shell Ca on Preventing of Bone Loss after Ovariectomy

Qualidade de ovos de codornas alimentadas com dietas que contém mananoligossacarídeos e níveis reduzidos de cálcio

. It was evaluated the effects of diets containing mannan oligosaccharides (MOS) and reduced calcium (Ca) levels on Japanese quail eggs quality. Two hundred quails, at 40 days of age, were used during three periods of 28 days each one. The experimental design was entirely randomized in factorial arrangement 2 x 2 + 1 (Ca x MOS levels and a control diet), with four replicates. The treatments were Ca (2.2 and 1.9%) and MOS (0 and 0.1%) levels and a control diet (with no MOS and with 2.5% Ca). It was studied the internal quality (weight and index of yolk and albumen and Haugh unit) and external (eggshell weight, Ca content, and thickness and specific gravity) of quail eggs. There was no effect (P>0.05) of the treatments on the weight and index of yolk and albumen, Haugh unit and eggshell weight and thickness. However, MOS inclusion increased (P<0.02) Ca content on the eggshell in 2.75% and the reduction of Ca levels decreased (P<0.01) the specific gravity of eggs from quails fed diets containing 1.9% of Ca. It was concluded that diets with 1.9% of Ca and supplemented with 0.1% of MOS can be used by Japanese quails with no negative effects on the internal and external quality of the eggs.

Fabrication, development and characterisation of calcium phosphate based bioceramic coatings on 316L stainless steel for biomedical applications

Hydroxyapatite (HAP) was synthesised by a precipitation method, using naturally available chicken eggshell (Ca source) and phosphoric acid (P source). The synthesised powder was electrophoretically deposited on 316L stainless steel (316L SS). The chemical composition of the synthesised powder was analysed. Ca was estimated by conventional EDTA titration and P by a gravimetric method. The Ca/P ratio was found to be 1·67, which resembles the stoichiometric HAP ratio. Fourier transform infrared (FT-IR) and powder X-ray diffraction (XRD) techniques were employed to confirm HAP formation. FT-IR spectra showed the characteristic peaks for phosphate and hydroxyl groups. The XRD results revealed that the major characteristic peaks of HAP appear in the regions of approximately 26°, 28°, 29°, 30–35°, 39°, 46°, 49° and 50° (2θ) and also indicated that no impurities are formed during HAP formation. The in vitro evaluation of HAP coated 316L SS was performed in simulated body fluid (Ringer’s solution) of pH 7·4 at 37°C in a thermostatic water bath through cyclic polarisation, and the results indicated that the HAP coated 316L SS has better corrosion resistance than uncoated 316L SS.

Hen's eggshell calcium

In Japan, insufficient calcium (Ca) intake is serious problem for health which may be associated with the high prevalence of osteoporosis among the aged. The intake of most nutrients has been sufficient, however, the Ca intake has never been sufficient. Eggshell Ca has as much as 38% of Ca and low phosphorus content. Eggshell Ca was more soluble than Ca carbonate and was as much as milk products. Eggshell Ca has been shown to exhibit higher absorptivity and availability than Ca carbonate. Furthermore, it has been reported that eggshell Ca is more effective in increasing bone mineral density in ovariectomized osteoporotic rats. These results suggest that eggshell Ca could be beneficial for bone and we propose Ca fortified foods which contain eggshell Ca as a nutraceutical.

Effect of 1alpha-hydroxyvitamin D3 and egg-shell calcium on bone metabolism in ovariectomized osteoporotic model rats.

Egg-shell calcium (Ca) is one of the effective Ca sources for bone metabolism. In the present study, we investigated whether egg-shell Ca had similar effects compared with calcium carbonate (CaCO3) when vitamin D3 (1alpha(OH)D3) treatment was given to an osteoporotic rat model. In both 1alpha(OH)D3-supplemented and -unsupplemented rats, the bone mineral density (BMD) of the lumber spine in the vitamin-supplemented group increased significantly compared with the unsupplemented group. In a Ca balance study, there were also significant differences in intestinal Ca absorption, urinary Ca and fecal Ca between the vitamin-supplemented and -unsupplemented groups. These results show that egg-shell Ca could have similar effects to CaCO3 on bone metabolism. In contrast with CaCO3, vitamin D3 supplementation did not significantly increase serum Ca levels in the egg-shell Ca group; however, the mechanism of Ca absorption is still unclear. Our results suggest that egg-shell Ca may be an effective nutrient in Ca metabolism for people treated with vitamin D3.

Enhancement of Plasma Calcium Concentration by Dietary Calcium Sources and Oligosaccharides in Neonatal Chicks

Yamashita, T., Tobisa, M., Ohgushi, A. and Furuse, M. 2001. Enhancement of plasma calcium concentration by dietary calcium sources and oligosaccharides in neonatal chicks. J. Appl. Anim. Res., 19: 25–32. The effects of dietary calcium (Ca) sources and several oligosaccharides on plasma Ca concentration were examined in the neonatal cluck. In the first experiment, the efficiency of Ca carbonate and eggshell Ca teas compared by determining plasma Ca concentration at 15, 30, 60 and 120 min post oral administration. It was significantly higher in the eggshell Ca treatment than in the Ca carbonate treatment and the peak for plasma Ca concentration was obtained at 30 min. The effect of dietary oligosaccharides on plasma Ca concentration was investigated by nsmg isomaito-oligosaccharides, gentiooligosaccharides and nigerooligosaccharides with Ca carbonate. In the second experiment Nigerooligosaccharides enhanced plasma Ca concentration when compared with other treatments. The efficacy of lactose and nigerooligosaccharides on plasma Ca concentration was compared in the third experiment, but no significant, difference was detected between them. These results suggested that plasma Ca concentration of the neonatal chick can be modified, by dietary Ca sources and the supplementation with specific oligosaccharides.

Calcium uptake by chorioallantoic membrane: effects of vitamins D and K

The chorioallantoic membrane (CAM) of birds is an epithelial tissue that actively transports large amounts of Ca during embryonic development. In this study the effect of vitamins D and K on Ca uptake by the CAM was studied. Four dietary treatments were used to produce eggs that are the following: deficient in vitamins D and K (-D/-K), sufficient in both (+D/+K), or deficient in one and sufficient in the other (-D/+K or +D/-K). Vitamin D-deficient (-D) Japanese quail embryos (from hens fed 1,25-dihydroxyvitamin D3) do not hatch because of severe Ca deficiency resulting from their inability to obtain Ca from shell, whereas vitamin K deficiency results in only 14% reduction in hatchability. The results demonstrate that Ca uptake by CAM is vitamin D dependent and only slightly vitamin K dependent. Ca-binding activity of CAM extracts was unchanged by vitamin K deficiency, and only a small increase was provided by vitamin D treatment. Vitamin D stimulated both Ca entry and exist from the chorion cells as indicated by the increased accumulated 45Ca in +D embryos. We conclude that vitamin D is essential for the utilization of eggshell Ca by the developing embryo and hence its survival, suggesting that Ca transport across the CAM is largely a vitamin D-dependent process.

Egg-shell calcium and the hatching of Globodera rostochiensis

The Ca 2+ content of Globodera rostochiensis egg-shells was investigated by X-ray microanalysis. When intact eggs were treated with 5 mM sodium 1, 2-di (2-aminoethoxy) ethane- N,N,N',N',-tetra-acetate (EGTA) only part of the egg-shell Ca 2+ was removed. Similarly treated opened egg-shells lost almost all their Ca 2+. We think that Ca 2+ of intact egg-shells which is accessible to EGTA is in the outer part of the shell and that the inaccessible Ca 2+ is in the inner lipoprotein layer. Much Ca 2+ was removed from opened egg-shells by the hatching agents Zn 2+, La 3+ and decationised potato-root exudate, but none by dilute acetic acid or Mg 2+. Hatching agents, by binding to or replacing Ca 2+, may change lipoprotein membrane structure. Eggshells treated with potato-root exudate contained about 3 times as much Ca 2+ as untreated shells, because the treatment makes additional binding sites available. Our results suggest that three types of Ca 2+ -binding sites occur in the egg-shell.