The potential of lemon peel powder as an additive in layer quails (Coturnix coturnix Japonica): An experimental study.

The current research intended to examine the impact of dietary lemon peel powder (LPP) on laying quail performance, egg quality criteria, and the antioxidant capacity of the yolk. A total of 120 female Japanese quails (272.6±9.3 g), aged 21 weeks, were allotted to 6 trial groups, each with 5 replicates of 4 quails. Additions of 0, 1, 2, 3, 4, or 5 g/kg of LPP to the basal diet were used to create the treatment groups. Quails were fed ad libitum for 70 days. Neither performance parameters nor egg production was affected by LPP. However, eggshell-breaking strength improved by adding 2 g/kg LPP to the diet, but worsened at 5 g/kg. Moreover, the relative weight of eggshell and yolk L* value decreased with the treatments. Dietary LPP enhanced oxidative stability, reducing malondialdehyde (MDA) and increasing 1,1-diphenyl-2-picrylhydrazyl (DPPH) yolk values. The current study demonstrated that LPP, a safe and easily accessible agricultural by-product, enhanced eggshell quality when it was included in the diet of laying quails at doses of 2 g/kg. In contrast, improvement of yolk antioxidant capacity required increased amounts of LPP (4 g/kg). LPP could be advantageous to animal nutrition as an adequate substitute to reduce waste by-products.

Synthesis and Characterization of Nano-Hydroxyapatite Obtained from Eggshell via the Hydrothermal Process and the Precipitation Method.

Hydroxyapatite (HA) is a major component of the inorganic minerals in the hard tissues of humans and has been widely used as a biomedical ceramic material in orthopedic and dentistry applications. Because human bone contains several impurities, including carbonates, chlorides, fluorides, magnesium, and strontium, human bone minerals differ from stoichiometric HA. Additionally, natural bone is composed of nano-sized HA, and the nanoscale particles exhibit a high level of biological activity. In this paper, HA is prepared via the hydrothermal process because its reaction conditions are easy to control and it has been shown to be quite feasible for large-scale production. Therefore, the hydrothermal process is an effective and convenient method for the preparation of HA. Furthermore, eggshell is adopted as a source of calcium, and mulberry leaf extract is selectively added to synthesize HA. The eggshell accounts for 11% of the total weight of a whole egg, and it consists of calcium carbonate, calcium phosphate, magnesium carbonate, and organic matter. Eggshell contains a variety of trace elements, such as magnesium and strontium, making the composition of the synthesized HA similar to that of the human skeleton. These trace elements exert considerable benefits for bone growth. Moreover, the use of eggshell as a raw material can permit the recycling of biowaste and a reduction in process costs. The purpose of this study is to prepare HA powder via the hydrothermal method and to explore the effects of hydrothermal conditions on the structure and properties of the synthesized HA. The room-temperature precipitation method is used for the control group. Furthermore, the results of an immersion test in simulated body fluid confirm that the as-prepared HA exhibits good apatite-forming bioactivity, which is an essential requirement for artificial materials to bond to living bones in the living body and promote bone regeneration. In particular, it is confirmed that the HA synthesized with the addition of the mulberry leaf extract exhibits good in vitro biocompatibility. The morphology, crystallite size, and composition of the carbonated nano-HA obtained herein are similar to those of natural bones. The carbonated nano-HA appears to be an excellent material for bioresorbable bone substitutes or drug delivery. Therefore, the nano-HA powder prepared in this study has great potential in biomedical applications.

Risk assessment of Artemia egg shell-Mg-P composites as a slow-release phosphorus fertilizer during its formation and application in typical heavy metals contaminated environment.

Artemia egg shell loaded with nano-magnesium (shell-Mg) can be used to recover phosphorus from wastewater. The exhausted Artemia egg shell-Mg (denoted as shell-Mg-P) can be used as a slow-release fertilizer for phosphorus reuse. However, due to the coexistence of heavy metal ions in the environment, the application of slow-release fertilizer for phosphorus removal and reuse may have potential risks. In this paper, the potential risks of Pb2+, Cd2+, Zn2+ and Cu2+ in phosphorus wastewater and soil were studied from the formation and application process of shell-Mg-P. The result showed that shell-Mg adsorbed Pb2+, Cd2+, Zn2+ and Cu2+ in phosphate wastewater during the formation of shell-Mg-P and became shell-Mg-P-metal hybrid biomaterial. Although the experiment proved that the existence of heavy metal ions did not affect the phosphorus slow-release behavior of slow-release fertilizer, but the heavy metal ions in the shell-Mg-P-metal were also slow released. The pot experiment results confirmed that the slow-release phosphorus fertilizers (shell-Mg-P and shell-Mg-P-metal) in the soil polluted in low concentration of heavy metals can reduce the amount of heavy metals in whole wheat seedlings and promote wheat seedling growth. However, the application of slow-release fertilizers increased the translocation efficiency (TFR to SL) of metal from root (R) to aboveground part (stem and leaves, SL), promoted the transportation of heavy metals from roots to the stems and leaves, and increased the safety risk of the wheat seedling edible. Therefore, besides the positive role of slow-release fertilizers in retaining heavy metals and reducing the amount of heavy metals in whole seedlings, the risk that it may aggravate the translocation of heavy metals to stems and leaves should be paid more attention, so as to ensure the safe and reliable application of slow-release fertilizers.

Paper-based dots and smartphone for detecting counterfeit country eggs.

Herein, we present for the first time, the employment of paper-based devices for rapidly differentiating original country eggs from the plain broiler eggs that have been coated with tea to appear as the former. The devices leverage two types of phenomena involving the phenols present in tea in precisely 5 min, namely precipitation, which produces a well-defined dark bluish precipitate on the surface of the counterfeit country eggs or tea-coated broiler eggs and de-coloration, wherein the dried layer of tea coating present on the surface of the dummy country eggs get dissolved, thereby revealing the white colour of the plain broiler egg shell. To reduce the subjectivity, a smartphone application 'Eggo' has been developed which is capable of detecting the spots produced by both the methods using mobile's camera. Fourier Transform Infrared Spectroscopy (FTIR) analysis was performed to study the changes occurring on the shell surface. Such sophisticated yet simple technologies will revolutionize food fraud analysis.

Imaging of eggshells and eggs in the gastrointestinal tract: pictorial essay.

Ingestion of eggshell in its natural form or as ground and powdered product has become a popular means of dietary calcium supplementation in adults. These substances appear as conspicuous radiopaque material within the gastrointestinal tract on radiographs or computed tomography of the abdomen. The ingested eggshell fragments are usually visible as curvilinear structures on profile view, whereas the ground or powdered eggshells appear as granular densities. This article illustrates the spectrum of findings that are observed following eggshell ingestion by patients undergoing radiological evaluation for various unrelated medical conditions. Potential complications of eggshell consumption are discussed, and two cases of intra-rectal egg insertion for palliative relief of pelvic pain are presented.

Eggshell based biochar for highly efficient adsorption and recovery of phosphorus from aqueous solution: Kinetics, mechanism and potential as phosphorus fertilizer.

Development of an efficient and green adsorbent is of great significance for phosphorus removal and recovery from eutrophic water. This work prepared an eggshell modified biochar (ESBC) by co-pyrolysis of eggshells and corn stalk. ESBC exhibited an excellent performance for phosphorus adsorption over a wide pH range (5-13), and achieved a maximum adsorption of 557.0 mg P/g. The adsorption process was well fitted by pseudo-second-order model (R2 > 0.962) and Sips model (R2 > 0.965), and it was endothermic (ΔH0 > 0) and spontaneous (ΔG0 < 0) according to thermodynamic analysis. The column experiment confirmed the feasibility of ESBC as a filter media for phosphorus removal in flow condition, and obtained a P removal of 460.0 mg/g. Soil burial tests indicated P-laden ESBC has a good P slow-release performance (maintained for up to 25 days). Overall, ESBC has a promising application potential as an efficient adsorbent for phosphorus recovery and subsequently as a slow-release fertilizer.

Influence of dietary dacitic tuff breccia on laying hen performance and egg quality parameters and bone structure at 85 weeks of age after a non-anorexic molt program at 73 to 77 weeks.

The objective of the study was to assess the efficacy of AZOMITE (AZM), a dacitic tuff breccia, in laying hens through egg quality and production parameters. A total of ninety six 73-wk-old Hy-Line W-36 commercial laying hens were randomly assigned to 2 dietary treatments, a control diet and the same diet containing 0.25% AZM, with 24 replicates of 2 hens/replication. From 73 to 77 wk, hens went through nonanorexic molt, and, from 77 to 85 wk, the hens were evaluated for egg production, eggshell quality, and bone health. At wk 85, tibiotarsi were collected for ash and mineral composition, ileal contents were collected for calcium, phosphorus, apparent metabolizable energy corrected for N (AMEn), and apparent nitrogen retention (ANR) evaluation. AZM-fed hens tended to have higher body weight (P = 0.07) from 82 to 83 and 84 to 85 wk, and higher hen day egg production than control (90.54 vs. 79.51%, P = 0.005) from 84 to 85 wk. In general, no differences were reported in feed intake, eggshell color, egg weight, albumen height, Haugh units, or eggshell thickness (P > 0.05). However, shell strength and elasticity were improved (P < 0.02) and yolk color was decreased (P = 0.03) in AZM-fed hens than control. Moreover, the digestibility of Ca, AMEn, and ANR was increased with 0.25% AZM compared to control (P < 0.01). Tibiotarsi P and Ca percentage were lower in AZM-fed birds than control (P < 0.01), without affecting bone strength and mineral density (P > 0.36). Therefore, the use of 0.25% AZM showed a potential in improving egg production and eggshell strength, while maintaining bone quality in post-molt laying hens.

Valorization of potato peels and eggshells wastes: Ca-biocomposite to remove and recover phosphorus from domestic wastewater.

Potato peel (PP) waste are generated in huge quantities, causing environmental pollution and health problems. Therefore, obtaining value-added products from PP is a current research challenge. In this work, novel Ca-biocomposites for phosphorus (P) removal were prepared by pyrolysis (500-800 °C) using eggshell (ES) and PP (ES/PP = 1:2 ratio by weight). ESPP-700 (pyrolyzed at 700 °C), reached a Qmax of 174.8 mg P/g, while the application of Ca-biocomposites in domestic wastewater showed 85.96% of P removal. According to the pseudo-second-order kinetic model, P adsorption was dominated by chemisorption, follows by apatite precipitation. The P solubility (62.5 wt.%) in formic acid (2.0 wt.%) and the water-soluble P (3.2 wt.%) for ESPP-700 after P adsorption, indicated that the final product would work as fertilizer for acidic soils. This is an important step in the management of agricultural wastes to implement the 3R slogan "Reduce, Reuse, Recycle" towards a circular economy.

Restoring phosphorus from water to soil: Using calcined eggshells for P adsorption and subsequent application of the adsorbent as a P fertilizer.

This study investigated the solution for two environmental issues: excess of P in water and its deficiency in soil, which is restored by transferring the adsorbed P from water into the soil using eggshell as an adsorbent. The eggshells were calcined at different temperatures to improve their adsorption capacity, and evaluated for their physical/chemical properties and P adsorption capacity. The eggshells calcined at 800 °C (CES-800) had the highest P adsorption; CaCO3 decomposed into 23.6% of CaO and 40.8% of Ca(OH)2, eluting more Ca that reacted with soluble P in water. X-ray diffraction analysis confirmed that CES-800 removed P as hydroxylapatite by reacting with Ca. Pseudo-first-order and Langmuir models suitably described the kinetic and equilibrium of P adsorption by CES-800, respectively. The maximum adsorption capacity of CES-800 was 108.2 mg g-1. As the solution pH increased from 3 to 11, the adsorption amount decreased from 99.8 mg g-1 to 62.3 mg g-1. The feasibility of CES-800 as a filter medium was assessed using real lake water under dynamic flow conditions; > 90% of P removal was achieved at 158 h, and the P adsorbed was 11.5 mg g-1. When CES-800 and P adsorbed CES-800 (P-CES-800) were applied to the soil at the studied rates, the earthworms were unaffected by toxicity, suggesting the use of both adsorbents in soil without adverse effects. The shoot fresh weight, tiller number, and total dry weight significantly increased in P-CES-800 applied rice plants compared to the control plants, indicating that P-CES-800 can be a good alternative to conventional P-fertilizer in rice cultivation.

Effects of Avian Eggshell Oiling With Diluted Bitumen Show Sublethal Embryonic Polycyclic Aromatic Compound Exposure.

Breeding birds that become oiled may contaminate the shells of their eggs, and studies of conventional crude oil suggest that even small quantities can be absorbed through the eggshell and cause embryotoxicity. Unconventional crude oils remain untested, so we evaluated whether a major Canadian oil sands product, diluted bitumen (dilbit), would be absorbed and cause toxicity when applied to eggshells of two species, domestic chicken (Gallus gallus domesticus) and double-crested cormorant (Nannopterum auritum). We artificially incubated eggs and applied lightly weathered dilbit (Cold Lake blend) to the eggshells (0.015-0.15 mg g-1 egg in chicken; 0.1-0.4 mg g-1 egg in cormorant) at various points during incubation before sampling prehatch embryos. Polycyclic aromatic compound (PAC) residue in cormorant embryos was elevated only at the highest dilbit application (0.4 mg g-1 egg) closest (day 16) to sampling on day 22. In contrast, cormorant liver cytochrome P450 1a4 (Cyp1a4) mRNA expression (quantitative polymerase chain reaction assay) was elevated only in embryos treated with the earliest and lowest dilbit application (0.1 mg g-1 egg on day 4). These results confirm that dilbit can cross through the eggshell and be absorbed by embryos, and they imply rapid biotransformation of PACs and a nonmonotonic Cyp1a4 response. Despite evidence of exposure in cormorant, we found no detectable effects on the frequency of survival, deformity, and gross lesions, nor did we find effects on physiological endpoints indicative of growth and cardiovascular function in either chicken or cormorant. In ovo dilbit exposure may be less toxic than well-studied conventional crude oils. The effects of an oil spill scenario involving dilbit to bird embryos might be subtle, and PACs may be rapidly metabolized. Environ Toxicol Chem 2022;41:159-174. © 2021 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Valorization of bio-waste eggshell as a viable source of dietary calcium for confectionery products.

BACKGROUND: The market of innovative foods is developing very dynamically and consumers are paying increasingly more attention to the health properties of the food offered. In addition, many studies on food consumption indicate a deficiency of minerals in the diet and experts suggest that the consumption of fortified functional foods in a selected group of consumers may contribute to the improvement of their health and quality of life. RESULTS: The aim of the work was to present the possibility of using bio-waste eggshell as a source of calcium in designing confectionery products. A snack with high calcium (0.567 g Ca kg-1 of product) and fiber (11.44 g kg-1 of product) content and potentially high calcium bioavailability was prepared. The results of the sensory expert-based assessment indicate a high (6-7 conventional units) overall sensory quality (OSQ) of the four flavor versions of the product during up to 3 months storage. In order to accurately illustrate the effect of storage time on the quality of the samples, texture analysis was carried out and the presented results indicate low correlations (r = -0.5554; r = -0.1494) between instrumentally measured hardness and sensory attribute of crispness, as well as hardness and OSQ, respectively. CONCLUSION: It is possible to reuse bio-waste eggshells as a calcium-fortifying substance for food design. The addition of powdered eggshells to food is an interesting direction, both for nutritional reasons (high concentration of calcium and optimal bioavailability) and also for environmental protection (reduction of post-production waste). © 2021 Society of Chemical Industry.

Effect of organic and inorganic dietary selenium supplementation on gene expression in oviduct tissues and Selenoproteins gene expression in Lohman Brown-classic laying hens.

BACKGROUND: The oviduct of a hen provides a conducive environment for egg formation, which needs a large amount of mineral elements from the blood via trans-epithelial permeability. Eggshell is the calcified layer on the outside of an egg that provides protection and is critical for egg quality. However, little is known about the genes or proteins involved in eggshell formation, and their relationship to dietary microminerals. We hypothesized that dietary selenium supplementation in chickens will influence genes involved in eggshell biomineralization, and improve laying hen antioxidant capacity. The objective of this research was to investigate how organic and inorganic dietary selenium supplementation affected mRNA expression of shell gland genes involved in eggshell biomineralization, and selenoproteins gene expression in Lohman Brown-Classic laying hens. RESULTS: Shell gland (Uterus) and liver tissue samples were collected from hens during the active growth phase of calcification (15-20 h post-ovulation) for RT-PCR analysis. In the oviduct (shell gland and magnum) and liver of laying hens, the relative expression of functional eggshell and hepatic selenoproteins genes was investigated. Results of qPCR confirmed the higher (p < 0.05) mRNA expression of OC-17 and OC-116 in shell gland of organic Se hen compared to inorganic and basal diet treatments. Similarly, dietary Se treatments affected the mRNA expression of OCX-32 and OCX-36 in the shell gland of laying hens. In the magnum, mRNA expression of OC-17 was significantly (p < 0.05) higher in hens fed-bacterial organic, while OC-116 mRNA expression was down-regulated in dietary Se supplemented groups compared to non-Se supplemented hens. Moreover, when compared to sodium selenite, only ADS18 bacterial Se showed significantly (p < 0.05) higher mRNA levels in GPX1, GPX4, DIO1, DIO2 and SELW1, while Se-yeast showed significantly (p < 0.05) higher mRNA levels in TXNRD1 than the non-Se group. CONCLUSIONS: Dietary Se supplementation especially that from a bacterial organic source, improved shell gland and hepatic selenoproteins gene expression in laying hens, indicating that it could be used as a viable alternative source of Se in laying hens. The findings could suggest that organic Se upregulation of shell gland genes and hepatic selenoproteins in laying hens is efficient.

Effects of Eggshell Membrane on Keratinocyte Differentiation and Skin Aging In Vitro and In Vivo.

Skin aging is one of the hallmarks of the aging process that causes physiological and morphological changes. Recently, several nutritional studies were conducted to delay or suppress the aging process. This study investigated whether nutritional supplementation of the eggshell membrane (ESM) has a beneficial effect on maintaining skin health and improving the skin aging process in vitro using neonatal normal human epidermal keratinocytes (NHEK-Neo) and in vivo using interleukin-10 knockout (IL-10 KO) mice. In NHEK-Neo cells, 1 mg/mL of enzymatically hydrolyzed ESM (eESM) upregulated the expression of keratinocyte differentiation markers, including keratin 1, filaggrin and involucrin, and changed the keratinocyte morphology. In IL-10 KO mice, oral supplementation of 8% powdered-ESM (pESM) upregulated the expression of growth factors, including transforming growth factor ß1, platelet-derived growth factor-ß and connective tissue growth factor, and suppressed skin thinning. Furthermore, voltage-gated calcium channel, transient receptor potential cation channel subfamily V members were upregulated by eESM treatment in NHEK-Neo cells and pESM supplementation in IL-10 KO mice. Collectively, these data suggest that ESM has an important role in improving skin health and aging, possibly via upregulating calcium signaling.

Formulation and Characterization of Calcium-Fortified Jelly and Its Proximate Composition and Sensory Analysis.

Calcium is a dynamic mineral. Recent discoveries designate that low intake of calcium generates deficiencies and path to other diseases. Food fortification could play a key role to overcome this problem. To cope with this deficiency problem, jellies were formulated with food-grade calcium salts and chicken eggshell powder. In the present study, three different concentrations of calcium salts, as well as eggshell powder were used to formulate jellies. The results of the sensory evaluation indicated that the two jelly products (A&D) in the current study were suitable for consumers. Results of Atomic Absorption Spectrophotometer revealed Jelly A and jelly D had 151±0.05 ppm and 133±0.06 ppm calcium concentration, respectively. Proximate analysis of Jelly A showed that it has 6.0±0.01% ash, 9.2±0.1% moisture, 0.4±0.01 g crude protein, 82.79±0.001 g crude fiber, and 0.61±0.001 g crude fat, while the jelly D that was made with chicken eggshell powder exhibited 6.0±0.01% ash, 10.1±0.1% moisture, 0.5±0.01 g protein, 84.54±0.01 g crude fiber and 1.61±0.01 g crude fat. Therefore, these two jelly A & D were greatly appreciated among other attributes. In spite of naturally available calcium-rich sources, calcium-fortified jellies can be consumed by individuals who are incapable to take sufficient calcium from their diet.

Facile carboxylation of natural eggshell membrane for highly selective uranium (VI) adsorption from radioactive wastewater.

With the commercial nuclear technology rising in society nowadays, it is of paramount importance to remove uranium (VI) in radioactive wastewater through a cost-effective and efficient way. Due to simple operation, low cost and abundant adsorbents, the adsorption method has been widely used to treat the radioactive wastewater. However, unsatisfactory selectivity and potential secondary pollution of most adsorbents hamper their practical large-scale application. To overcome these limitations, an effective and green absorbent is developed by functionalizing the waste eggshell membrane (ESM) with carboxyl-rich agents. This design concept transfers waste ESM (or "trash") into a unique "treasure" absorbent for directly handling radioactive wastewater. The resultant ESM-COOH shows excellent adsorption selectivity toward uranium (VI) with the selectivity coefficient of 75%, exceeding a majority of reported adsorbents. Moreover, its adsorption capacity still maintains 84% of the initial value after six cycles, suggesting good reusability. These excellent features enable the ESM-COOH to adsorb uranium (VI) highly selectively and efficiently. This work offers a concept to transfer biological wastes into treasure for the mass remediation of water body.

Early, intensive marine resource exploitation by Middle Stone Age humans at Ysterfontein 1 rockshelter, South Africa.

Modern human behavioral innovations from the Middle Stone Age (MSA) include the earliest indicators of full coastal adaptation evidenced by shell middens, yet many MSA middens remain poorly dated. We apply 230Th/U burial dating to ostrich eggshells (OES) from Ysterfontein 1 (YFT1, Western Cape, South Africa), a stratified MSA shell midden. 230Th/U burial ages of YFT1 OES are relatively precise (median ± 2.7%), consistent with other age constraints, and preserve stratigraphic principles. Bayesian age-depth modeling indicates YFT1 was deposited between 119.9 to 113.1 thousand years ago (ka) (95% CI of model ages), and the entire 3.8 m thick midden may have accumulated within ∼2,300 y. Stable carbon, nitrogen, and oxygen isotopes of OES indicate that during occupation the local environment was dominated by C3 vegetation and was initially significantly wetter than at present but became drier and cooler with time. Integrating archaeological evidence with OES 230Th/U ages and stable isotopes shows the following: 1) YFT1 is the oldest shell midden known, providing minimum constraints on full coastal adaptation by ∼120 ka; 2) despite rapid sea-level drop and other climatic changes during occupation, relative shellfish proportions and sizes remain similar, suggesting adaptive foraging along a changing coastline; 3) the YFT1 lithic technocomplex is similar to other west coast assemblages but distinct from potentially synchronous industries along the southern African coast, suggesting human populations were fragmented between seasonal rainfall zones; and 4) accumulation rates (up to 1.8 m/ka) are much higher than previously observed for dated, stratified MSA middens, implying more intense site occupation akin to Later Stone Age middens.

Application of response surface methodology for uranium(VI) adsorption using hydroxyapatite prepared from eggshells waste material: study of influencing factors and mechanism.

Hydroxyapatite (HAp) was synthesized from biowaste hen eggshells by wet precipitation method in which calcium hydroxide and phosphoric acid were used as precursors. The effectiveness of uranium(VI) adsorption onto HAp was investigated by batch adsorption experiments from aqueous solutions. The obtained HAp powder was characterized by X-ray diffraction, Fourier transform infrared spectroscopy, Raman spectrometry, point of zero charge and Scanning electron microscope. The factors and levels used during the experiments were pH (2-5), adsorbent mass (0.01-0.05 g), and initial U(VI) concentration (100-310 mg L-1). A Box-Behnken design combined with analysis of variance was used to interpret the main effect influencing the adsorption. The results showed that pH was the most significant parameter affecting U(VI). The kinetic data correlates well with the pseudo-second-order model. The adsorption isotherms fitted the Langmuir-1 type model with the qmax = 175.22 mg g-1 at 25 °C. The calculated value of the mean free energy indicates the chemisorption process. Under optimal conditions, the uranium effluent derived from the precipitation of ammonium uranyl carbonate removal performance of 98% was achieved. This study proved that HAp prepared from eggshell was an ecofriendly and low-cost adsorbent and was very effective for the adsorption of U(VI) from aqueous solutions.

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.

Preparation of Sterile Raw Material - Chicken Eggshells in the Process of their Transformation into Selected Calcium Salts.

BACKGROUND: The chicken eggshells and their subcrustal membranes are a valuable source of calcium, but they are not further processed but disposed of as waste from the food industry. Chicken eggshells have high content (>95%) of calcium carbonate. Some properties suggest that eggshells may be a promising alternative to the present calcium sources used in the pharmaceutical industry. METHODS: The effect of roasting chicken eggshells with a selected organic acid (citric or fumaric or lactic acid) on microbiological purity, including the presence of fungi and bacteria Salmonella spp., Staphylococcus aureus, Escherichia coli of obtained calcium salts, was investigated. In this study, chicken eggshells were subjected to chemical reactions with organic acids (citric, fumaric or lactic acid) at two different calcium-acid molar ratios (1:1 or 1:3) and the mixture was heat-treated for 1 or 3 hours at a temperature of 100°C or 120°C. RESULTS AND DISCUSSION: It was found that lactic acid was 100% effective against fungi, and the remaining citric and fumaric acids were -50% (regardless of the other examined conditions). The type of acid used has a significant effect on fungal growth inhibition (p<0.05). Fumaric acid and lactic acid will be nearly 100% effective against bacteria (100% fumaric acid and 97% lactic acid effectiveness), regardless of other factors. CONCLUSION: Lactic acid is the most effective against pathogenic flora - fungi and bacteria. The transformation of chicken eggshells into calcium lactate can provide us with sterile calcium salt, free of 100% fungi and 97% of all bacteria.

Improvement of eggshell quality by dietary N-carbamylglutamate supplementation in laying chickens.

Egg quality defects seriously reduce the quality grade and increase egg breakage in egg marketing activities. In this study, the effect of N-carbamylglutamate (NCG) on eggshell quality was investigated by evaluating calcium absorption and calcification in laying hens. A total of 30 newly hatched female Hy-Line chicks were randomly assigned to the control group (basal diet) and treatment group (basal diet supplemented with 1% NCG). At 25 wk, eggs from each group were obtained to assess egg quality parameters. Blood samples were collected for analysis of mineral, hormone, and amino acids levels at 16 h after laying egg. Uterine tissues were removed and fixed in 4% neutral paraformaldehyde solution or kept in liquid nitrogen for mineral determination, quantitative PCR, and Western blot. Results showed that the egg quality (eggshell thickness, strength and percentage, egg specific gravity, and eggshell effective thickness) was significantly increased while effective thickness of mastoid layer, width of mastoid gap, and mammillary knobs were significantly decreased by dietary NCG supplementation (P < 0.05). The levels of minerals (Ca, P, Fe, Mg, Na, K) in eggshell, plasma, and uterus were remarkably elevated (P < 0.05). Meanwhile, the concentrations of calcium metabolism-related hormones (17ß-estradiol, parathyroid hormone, and calcitonin) were increased in the NCG group (P < 0.05). Moreover, expression of calbindin 1, carbonic anhydrase 2, ovalbumin, ovotransferrin, ovocleidin-17, ovocleidin-116, and clusterin mRNAs, as well as calbindin 1 and ATP2A1 proteins in uterus, duodenum, and kidney, was all upregulated in hens fed with NCG (P < 0.05). In addition, the number of blood vessels in the uterus, height of uterine mucosal folds, villus length in endometrium, and areas of uterine mucosal folds were significantly increased in the NCG group (P < 0.05). In conclusion, dietary 1% NCG supplementation during 0 to 25 wk can improve eggshell quality through changes in endometrial morphology, expression of calcium metabolism-related genes, and secretion of related hormones to promote eggshell formation in the laying hens.