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This study was conducted to determine the influence of dietary glucosamine sulfate sodium (GSS) on laying performance, blood profiles, eggshell and inner quality of eggs and relative expression of the genes related to eggshell in laying hens at early stage. A total of 640 twenty-weeks-old Lohmann laying hens were randomly allotted to 4 treatments with 10 replicates of 16 hens each. The experiment lasted for 8 wk, and dietary treatments were: 1) CON, basal diet; 2) G1, CON + 0.2% GSS; 3) G2, CON + 0.4% GSS; 4) G3, CON + 0.6% GSS. The inclusion of GSS increased average daily feed intake, laying rate, and egg mass (P < 0.05) linearly during wk 21 to 25, 25 to 29, and 21 to 29, egg weight during wk 21 to 25 and 25 to 29, and improved (P < 0.05) feed conversion ratio linearly during wk 21 to 25. The supplementation of GSS increased (P < 0.05) albumen height quadratically, Haugh unit, calcium content, calcium mass, phosphorus content and phosphorus mass linearly at the end of 25th and 29th wk. At the end of 29th wk, the eggshell strength, eggshell weight, eggshell ratio, and eggshell thickness were increased (P < 0.05) linearly in GSS treatments compared with CON. The addition of GSS increased (P < 0.05) serum calcium, estrogen 2, and calcitonin, while decreased (P < 0.05) serum tartrate resistant acid phosphatase (TRAP), parathormone, IL-6 and prostaglandin E 2 (PGE 2 ) at the end of 29th wk. The inclusion of GSS increased (P < 0.05) the relative expression of ovocalyxin-32 and ovocalyxin-36 linearly at the end of 29th wk, and ovalbumin, osteopontin, calbindin 1, and ovocleidin-116 linearly at the end of 25th and 29th wk. Quadratic effects were observed (P < 0.05) in the laying rate during wk 21 to 25, serum TRAP and PGE 2 , the relative expression of ovocleidin-116 at the end of 29th wk. In summary, the inclusion of GSS up-regulated relative expression of osteopontin, ovocleidin-116, ovocalyxin-32 and ovocalyxin-36 in uterus, promoted the serum PGE 2 and calcitonin, thus increased the calcium content of eggshell and finally enhanced eggshell quality., Competing Interests: DISCLOSURES The authors declare no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Slightly acidic electrolyzed water (SAEW) is a safe and effective disinfectant, but its sterilizing efficiency is compromised by organic matter on the egg surface. Electrolyzed reduced water (ERW) is a harmless cleaner with a decontamination effect on a variety of surfaces and can be used to remove organic matter. This study assesses the effectiveness of a combination of ERW and SAEW in eliminating Salmonella and manure mixture from egg surfaces, as well as its impact on egg quality during storage. The results show that ERW (74.14%) was more effective than deionized water (DW, 64.69%) and SAEW (70.20%) (P < 0.05) in removing manure from egg surfaces. The damage to the cuticle of eggshell treated with ERW for 28 s was similar to that of DW (P > 0.05) and less than that of SAEW (P < 0.05). Spraying ERW for 10 s followed by SAEW for 18 s (ERW + SAEW) completely removed Salmonella from the egg surface, with no bacteria detected in the residual wash solution. Additionally, ERW + SAEW demonstrated superior preservation of egg quality during storage at 25℃ than the control and ERW single treatment (P < 0.05). Moreover, ERW + SAEW resulted in less weight loss compared to SAEW single treatment (P < 0.05). In conclusion, the sequential use of ERW and SAEW appears to be a promising approach for sterilizing eggs. It not only removes organic matter and Salmonella from the egg surface but also improves the preservation quality of the egg at 25 ℃., Competing Interests: DISCLOSURES The authors declare no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

The purpose of this study was to determine the effects of the dietary addition of zinc (Zn) in the form of sulphate (Zn-S), glycine (Zn-G), and hydroxychloride (Zn-H) on quail performance, eggshell quality, antioxidant status, mineral excretion, biomechanical properties and mineralization of bone. A total of 75 female quails (10-weeks-old) were randomly distributed into 3 groups with 5 replications, each with 5 female quails. Treatment groups were set up by adding Zn-S and Zn-H as the inorganic form and Zn-G as the organic form of zinc to the corn-soybean basal diet (34.14 mg/kg Zn) to obtain 50 mg/kg Zn and the feeding experiment lasted for 12 weeks. Performance parameters and egg production were not impaired by the Zn source (P > 0.05). The inclusion of Zn-S in the diet produced a reduction in eggshell thickness, while an improvement in yolk antioxidant capacity (measured as MDA concentration) was detected compared to the other Zn sources (P < 0.05). Shear strength and cortical bone area increased (P < 0.05) with Zn-G supplementation, however, the mineral concentration of the tibia was higher (P < 0.01) in those quails who had received Zn-H. Lower levels of mineral excretion were observed in both types of supplementations compared to Zn-S. Therefore, it can be stated that Zn-G or Zn-H supplementation in the diet of laying quails could be an interesting strategy to reduce mineral excretion and improve bone mineralization without affecting performance compared to Zn-S. However, further studies are needed to determine the differences between Zn-G and Zn-H., (© 2024. The Author(s).)

In this study, it was aimed to determine the effects of using UV light instead of formaldehyde in disinfection of hatching eggs on the microbial load, shell and yolk compositions, embryo development, hatching results and chick characteristics. 552 hatching eggs were divided into formaldehyde fumigation and disinfection with UV groups. After disinfection, bacteriological growth was not observed in groups. While the difference between the groups in terms of hatching results was insignificant, it was determined that the incubation period in the UV group was shorter than that of formaldehyde group. On the hatching day, relative chick weight and weight of bursa Fabricus were found to be higher in the UV group  . UV light can be used as an alternative to formaldehyde as disinfection of hatching eggs because it is easy to use and does not cause any negative effects on egg shell structure, embryo development, hatching results and chick characteristics.

A dose-response experiment was conducted to evaluate the effects of graded levels of dietary digestible threonine (dThr) during the first laying cycle on productive and reproductive performance, egg quality and immune responses of Japanese quail breeders (Coturnix coturnix japonica). Also, dThr requirements were determined based on nutrient dose-response data. A total of 450 (360 females and 90 males) 11-week-old breeders were allocated to five increment (+0.03%) levels of dThr (0.49%, 0.52%, 0.55%, 0.58%, 0.61% and 0.64%) with five replicates per treatment and 15 (12 females and three males) birds each. The experiment lasted for 12 weeks. In response to increasing dietary dThr levels, egg production, egg mass, feed efficiency, egg specific gravity, eggshell relative weight, eggshell thickness, egg fertility (EF) and immune response against sheep red blood cell (SRBC) inoculation were improved with quadratic trends and egg hatchability as set eggs was improved with linear trends. Japanese quail breeders fed a diet with 0.58% dThr concentration (threonine/lysine ratio of 59%) showed the productive performance traits, EF, eggshell quality and immune response against SRBC inoculation in the highest values. However, feed intake, egg weight, egg albumen and yolk relative weight, egg shape index, haugh unit and egg composition were not affected by increasing dietary dThr level. Based on the broken-line regression model, the dThr requirements to optimize productive performance, eggshell quality, EF and immune response against SRBC inoculation were estimated at 159-188, 169-183, 175 and 178 mg/bird per day, respectively. It is concluded, in the Japanese quail breeders during the first laying phase a daily dThr intake of 188 mg/bird, dietary dThr concentration at 0.58% (threonine/lysine ratio of 59%) is adequate for optimized productive and reproductive performance, eggshell quality and immune responses. The estimated requirements depend on what production parameter is taken into considered for optimization., (© 2024 Wiley‐VCH GmbH. Published by John Wiley & Sons Ltd.)

The higher availability of zinc (Zn) from organic than inorganic sources is already established, but more assertive and cost-friendly protocols on the total replacement of inorganic with organic Zn sources for laying hens still need to be developed. Because some discrepancy in the effects of this replacement in laying hen diets is noticeable in the literature, the objective of this meta-analysis was to properly quantify the effect size of total replacing inorganic Zn with organic Zn in the diet of laying hens on their laying performance, egg quality, and Zn excretion. A total of 2340 results were retrieved from Pubmed, Scielo, Scopus, WOS, and Science Direct databases. Of these, 18 primary studies met all the eligibility criteria and were included in this meta-analysis. Overall, the replacement of inorganic Zn with organic Zn, regardless of other factors, improved (p < 0.01) egg production by 1.46%, eggshell thickness by 0.01 mm, and eggshell resistance by 0.11 kgf/cm 2 . Positive results of the same nutritional strategy on egg weight and Zn excretion were only observed at specific conditions, especially when organic Zn was supplemented alone in the feed, not combined with other organic minerals. Therefore, there is evidence in the literature that the total replacement of inorganic Zn with organic Zn improves egg production, eggshell thickness, and eggshell resistance. Factors such as hen age and genetics, organic Zn source, concentration of Zn in the feed, and the strategy of its supplementation have to be more carefully considered in protocols designed to address egg weight and Zn excretion by the hen., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

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.

Background: Although hydrogen gas (H 2 ) has been widely used in industry and gradually used in medicine, it is less applied to agriculture, especially in modified atmosphere packing (MAP)., Results: During chicken egg storage, H 2 MAP not only slowed down or delayed the reduction in antioxidant capacities, but also alleviated the deterioration of egg quality. The extent of micro-cracks in the eggshell was also negatively influenced by H 2 , which might result in eggs being less vulnerable to microbes. The earlier results demonstrated that the H 2 MAP-extended shelf life of chicken eggs might be caused by the responses of eggshell and re-establishment of redox homeostasis. According to the data collected from different provinces of China, cost-economics analysis further suggested that the increase in the extra cost caused by H 2 was trivial compared with the original price of eggs., Conclusion: Together, H 2 MAP can prolong the shelf life of chicken eggs with the prospect of wider application. © 2021 Society of Chemical Industry., (© 2021 Society of Chemical Industry.)

The aim of the experiment was to evaluate the potential use of citric acid as a modifier of quality changes in table eggs during their storage. About 780 table hen eggs were collected on the same day. They were numbered individually and placed on trays 30 pcs on each. Control group (CA0) consisted of eggs unmodified with any additional substances. In experimental groups CA10 and CA15, eggshells were sprayed with the aqueous solution of citric acid (10 and 15% concentration, respectively). At the start of the experiment, only quality traits of eggs from the control group were analyzed. The remaining eggs were stored at 14°C and 70% RH (typical storage conditions). Their quality was evaluated after 7, 14, 21, and 28 d. The depth of the air cell, egg weight and specific gravity, traits of shell (permeability, strength, weight, thickness, density), and egg content (pH of yolk and albumen, Haugh units, yolk weight and color) were evaluated each time. The use of citric acid decreased the severity of qualitative changes. Citric acid-treated eggs demonstrated smaller weight loss, shallower air cell, higher structural albumen, less-intensive water diffusion from albumen to yolk indicating the improved resistance of the vitelline membrane. Owing to the fact that citric acid is accepted and recognized as a safe food preservative is a relatively cheap and available substance, it seems that it can be used to inhibit quality changes in table eggs during their storage., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Lower levels of calcium in adults increase the risk for osteoporosis, and in children, low calcium levels can impact their potential adult height. The study objective was to analyze the bioavailability and physicochemical properties of a calcium preparation based on chicken eggs. The base calcium preparation was enriched with one of a variety of biologically active substances, inter alia, vitamin D3, vitamin K, lysine, lactose, magnesium chloride and inulin. The newly developed calcium preparations were subjected to structural analysis using X-ray diffraction and scanning electron microscopy, and the hydrodynamic diameter for the molecules was determined using the dynamic light scattering method and their zeta potential. To determine the optimum storage conditions of calcium preparations, their hygroscopicity and bulk density were determined. The calcium preparations were also added to selected food products, such as apple juice with mango, fruit dessert (jelly) and beef meatballs. The enriched food products were subjected to sensory analysis. The study demonstrated the significant influence of additives to calcium preparation in terms of its hygroscopicity and morphology. It was found that all products with the addition of analyzed preparations were characterized by high sensory desirability. The results presented in the study comprise the basis for the development of new food products, enriched with calcium.

Following the accidental feeding of a compound feed containing the coccidiostat nicarbacin in layer breeder flocks (Lohmann Brown Classic), the birds displayed distinct clinical signs within a few hours. Mortality increased during the following 5 days, whereas laying performance and hatching rate of eggs during this period decreased markedly. Egg shell discoloration was observed as early as during the first day. As a consequence, an association between feeding of the coccidiostat nicarbacin and the observed symptoms was assumed. Recent studies indicate that Nicarbacin reduces the activity of aminolevulinic acid synthase type 1 (ALAS 1), which is responsible for the synthesis of protoporphyrin IX in the shell gland as main compound of brown egg shells. Reduced laying performance and increased mortality was likely due to nicarbacin-induced deregulated body temperature homeostasis and concomitant imbalances in acid-base status of the animals. The case reveals that the accidental feeding of nicarbacin to non-target animals such as laying hens and their parents may result in acute clinical symptoms. This highlights the necessity of appropriate care in handling feed additives and their premixes for specific non-target animals and should sensitize farmers and veterinarians., Competing Interests: Die Autoren erklären, dass kein Interessenkonflikt besteht., (Thieme. All rights reserved.)

Cage layer fatigue (CLF), which is commonly caused by calcium deficiency in the feed, leads to loss of structural bone and increase of bone fragility. In order to investigate the influence of low-calcium diets on bone quality and strength, histopathology, and egg quality, 72 laying hens were randomly allocated to 2 groups at 22 wk of age and received low calcium and control calcium until 34 wk, respectively. Egg production, feed consumption, BW, and egg quality were measured throughout. Bone mineral density, bone biomechanical properties, and histomorphology of femurs and tibias were assessed after birds were sacrificed in 26, 30, and 34 wk. The results showed that low-calcium (1.5%) diets decreased BW, feed consumption, and egg production. The broken eggs rate increased, and the eggshell strength and thickness were lower in treated birds than those in control birds at 30 wk and 34 wk. Femoral and tibial bone index and bone mineral density were lower, cortical thicknesses were thinner, and bone length were shorter over time when birds are in a low-calcium diet than those in control birds. In biomechanical properties, the values of stiffness, Young's modulus, and breaking strength were lower in both femurs and tibias in low-calcium hens at 30 wk and 34 wk than those in bones of control hens. In histomorphology of bone, the cortex turned thinner and there were more cavities in cortex and cancellous bone; the trabecular bone network was fewer, thinner, less cohesive, and generally fragmented; and trabeculae were less well-connected in low-calcium birds. Some cell nuclei in cancellous bone disappeared, and vacuolation was observed in bone cells. There appeared osteoid in cortex bone and cancellous bone in tibias. It was concluded that low-calcium diets could facilitate the development of osteoporosis characterized by an increase of osteoid and loss of structural bone and decrease the values of bone quality and strength, accompanied with a decrease in egg production and egg qualities, which may elucidate the developing mechanism of CLF., (Copyright © 2020. Published by Elsevier Inc.)

Biopolymers provide versatile platforms for designing naturally-derived wound care dressings through eco-friendly pathways. Eggshell membrane (ESM), a widely available, biocompatible biopolymer based structure features a unique 3D porous interwoven fibrous protein network. The ESM was functionalized with inorganic compounds (Ag, ZnO, CuO used either separately or combined) using a straightforward deposition technique namely radio frequency magnetron sputtering. The functionalized ESMs were characterized from morphological, structural, compositional, surface chemistry, optical, cytotoxicity and antibacterial point of view. It was emphasized that functionalization with a combination of metal oxides and exposure to visible light results in a highly efficient antibacterial activity against Escherichia coli when compared to the activity of individual metal oxide components. It is assumed that this is possible due to the fact that an axial p-n junction is created by joining the two metal oxides. This structure separates into components the charge carrier pairs promoted by visible light irradiation that further can influence the generation of reactive oxygen species which ultimately are responsible for the bactericide effect. This study proves that, by employing inexpensive and environmentally friendly materials (ESM and metal oxides) and fabrication techniques (radio frequency magnetron sputtering), affordable antibacterial materials can be developed for potential applications in chronic wound healing device area.

Objective: The use of low-energy electron irradiation for hatching egg disinfection was compared to formaldehyde fumigation., Material and Methods: To assess the efficacy, eggshells were examined for bacteria before and after disinfection. During incubation, hatching and fattening of 1400 Ross 308 broilers per group candling data, hatching rate, mortality rate and body mass development were recorded to evaluate the impact on health and performance. Additionally, data from the slaughter house and histological evaluation of organ samples were included in the health screening., Results: Regarding the efficacy, irradiation and formaldehyde significantly reduced the number of bacteria compared to non-disinfected eggs. However, no significant difference was recorded between both treatments. The irradiated group performed better than the reference group concerning the number of infertile eggs, early and late dead embryos and fertile eggs. The performance during fattening was nearly identical in both groups, while mortality differed slightly between the irradiated group and the group treated with formaldehyde. Upon slaughter, there were minor variations concerning the distribution of findings between the groups, but the number of anomalies was not higher than reference values suggest., Conclusion: In both groups no health problems were observed during rearing and the fattening performance met the targets of the manufacturer Aviagen for the used broiler line Ross 308., Clinical Relevance: Lower-energy electron irradiation proves to be an efficient and well-tolerated hatching egg disinfection procedure. Further field trials are necessary to establish low-energy electron irradiation as an alternative to formaldehyde fumigation., Competing Interests: The authors state that no competing interest was present during planning and conduction of the disinfection trial., (Thieme. All rights reserved.)

A study was conducted to evaluate the effect of dietary 25-hydroxyvitamin D 3 (25OHD) on pullet and egg-laying hen growth performance, egg production, and egg quality. Three hundred and ninety 1-day-old Hy-Line W36 pullets were randomly allocated to 3 treatments with 10 replicated cages and 13 birds per cage. Dietary treatments were vitamin D 3 at 2,760 IU/kg (D); vitamin D 3 at 5,520 IU/kg (DD), and vitamin D 3 at 2,760 IU/kg plus 25OHD at 2,760 IU (69 μg)/kg (25D). Body weight and feed intake were recorded at the end of each stage: starter 1 (0-3 wk), starter 2 (4-6 wk), grower (7-12 wk), developer (13-15 wk), prelay (15-17 wk), peaking (18-38 wk), layer 2 (39-48 wk), layer 3 (49-60 wk), layer 4 (61-75 wk), and layer 5 (76-95 wk). Egg production was recorded daily. Egg quality was evaluated every 8 wk starting from 25 wk. There was no difference in growth performance during the rearing period (0-17 wk). In the laying period (18-95 wk), DD showed lower feed intake at layer 2, but higher intake at layer 3 along with lower hen day production (HDP) from 22 to 48 wk compared to the other treatments. During the same period, the DD group laid smaller eggs with higher specific gravity and shell thickness compared with the other treatments or D alone at 40 wk, which may be partly due to the lower body weight. In contrast, 25D had better feed conversion ratio (feed intake per dozen of eggs) at layer 2, and higher overall (22-60 wk) HDP compared with DD. For the egg quality analysis, at 25 and 33 wk, both DD and 25D had higher Haugh unit compared with D. However, 25OHD has no effects on eggshell quality during the entire production period and no beneficial effects on egg production during the later laying period (after 60 wk). In summary, long-term and early supplementation of 25OHD has positive effects on egg production and egg quality, and the beneficial effects were mainly observed during the early laying stage., (Copyright © 2020. Published by Elsevier Inc.)

The present research was conducted to assess Mn requirements of broiler breeder hens. One hundred and twenty Cobb 500 hens, 22 wk of age, were individually allocated in cages. After fed a Mn-deficient diet (22.2 ppm), hens were randomly placed in treatments having 6 increments of 30-ppm Mn. All trace minerals were from laboratory grade sources being Mn from Mn sulfate (MnSO 4 H 2 O). Treatments were fed for 4 periods of 28 d. There were no interactions between dietary Mn and period for any evaluated response (P > 0.05). Requirements of Mn for hen day egg production and settable egg production were 115.8 and 56.6 ppm and 122.1 and 63.6 ppm (P < 0.05), respectively, using quadratic polynomial (QP) and broken line quadratic (BLQ) models, whereas total eggs and total settable eggs per hen had Mn requirements estimated at 115.7 and 56.6 and 121.8 and 61.7 ppm (P < 0.05), respectively. Number of cracked, defective, and contaminated eggs decreased, whereas hatchability, hatchability of fertile eggs, eggshell percentage, and eggshell palisade layer increased when hens were fed diets having 48.5 to 168.2-ppm Mn (P < 0.05). Maximum responses for egg weight and eggshell percentage were 117.7 and 63.6 ppm as well as 131.6 and 71.0 ppm (P < 0.05), respectively, using QP and BLQ models. Breaking strength and egg specific gravity had Mn requirements estimated at 140.2 and 112.7 ppm as well as 131.3 68.5 ppm (P < 0.05), whereas eggshell palisade layer and eggshell thickness were maximized with 128.8 and 68.8 ppm and 140.2 134.2 ppm, respectively, for QP and BLQ models (P < 0.05). Maximum yolk Mn content values were obtained using 118.0- and 118.4-ppm Mn by QP and BLQ models, respectively. The average Mn requirements estimated for QP and BLQ models is 128.4 and 92.3 ppm Mn (18.7 and 13.5 mg/hen/d), respectively, which is much lower than what has been currently recommended in commercial production., (Copyright © 2020. Published by Elsevier Inc.)

Exogenous phytase supplementation increases P and Ca availability to allow for the dietary reductions without negative consequences on productivity or skeletal health. Effects of a Buttiauxella sp. phytase (BSP) supplemented in available P (avP)-reduced and Ca-reduced diets on performance, BW, eggshell quality, serum biochemical bone markers, and bone densitometry were evaluated in egg-laying hens from 68 to 78 wk of age. One hundred hens were fed 1 of 5 diets (n = 20/treatment), including a positive control (PC) with 0.35% avP and 3.5% Ca, and the PC moderately reduced in avP and Ca levels by 0.187 and 0.159% of the diet (by 53 and 4.5%), respectively, (NC1) or severely reduced by 0.231 and 0.275% of the diet (by 66 and 7.9%), respectively, (NC2). Other diets were the NC1 or NC2 supplemented with BSP at 600 FTU/kg (NC1 + BSP or NC2 + BSP, respectively). Egg production and feed conversion ratio were maintained by NC1 but were 11.9% lower and 12.3% higher, respectively, with the NC2 than the PC, which was alleviated by supplemental BSP. Diet effects on FI and eggshell quality followed a similar pattern. Body weight was 2.9% lower for NC1, and 6.1% for NC2 than the PC; BSP alleviated the decreased BW. Serum pyridinoline (bone resorption marker) was 20 to 27% higher in NC2 hens than in the other groups, with no effects on other bone markers. Total and trabecular space bone mineral density in the proximal metaphysis were 8.4 and 15.2% lower for NC1, respectively, and 12.1 and 26.7% lower for NC2, respectively, than PC. Supplemental BSP completely alleviated the decreased bone densitometry measures in NC1, but only partially in NC2. The NC1 hens maintained performance but had decreased BW and bone quality; phytase supplementation restored productivity, BW, and bone quality. The Ca and avP deficiencies in the NC2 hens relative to other groups were partially alleviated by the 600 FTU/kg BSP., (Copyright © 2020. Published by Elsevier Inc.)

We investigated effects of rearing cage type and dietary limestone particle size (LPS) on egg production, egg weight, eggshell, and bone quality in laying hens. The pullets were reared in conventional (CON; 20 chicks/cage, 270 cm 2 /chick) or furnished (FUR; 30 chicks/cage; 636 cm 2 /chick) cages and fed 3 LPS (fine, <0.595 mm; medium, 0.595 to <1.68 mm; and 1:1 mixture of F and M wt/wt) to 16 wk of age (woa). Pullets were transitioned to laying furnished cages and retained rearing treatment combination identities (n = 5, 20 hens/cage). Hens had free access to common commercial layer diet and water through to 40 woa. Eggs were recorded daily for calculation of hen day egg production (HDEP). Subsamples of eggs laid on the first day of 24, 28, 32, 36, and 40 woa were used for eggshell quality analyses. Two hens per cage were sacrificed on the last day of 24 and 40 woa for femur and tibia quality assessments. There was no interaction (P > 0.05) between rearing cage type and dietary LPS on response variables. At 19 and 20 woa, HDEP was higher (P < 0.01) for FUR than CON reared hens but was similar (P > 0.05) afterward. At 40 woa, FUR reared hens had higher (P < 0.05) body weight (BW), egg weight (EW), eggshell thickness, and eggshell weight and tended (P < 0.10) to have higher femur and tibia mineral density (BMD) and mineral content (BMC) than CON reared hens. Rearing dietary LPS had no effect (P > 0.05) on HDEP, BW, EW, and eggshell quality. Although, rearing dietary LPS did not affect (P > 0.05) femur and tibia BMD and BMC; at 24 woa, hens reared on medium LPS tended to have higher femur BMD (0.17 vs. 0.14 g/cm 2 ; P = 0.079) and BMC (0.99 vs.0.78 g; P = 0.088) than hens reared on fine LPS. In conclusion, hens reared in furnished cages had better eggshell quality but had marginal effects on femur and tibia quality, whereas rearing dietary LPS had no effects on eggshell and bone attributes in hens., (Copyright © 2020. Published by Elsevier Inc.)

A 3 × 2 factorial arrangement of treatments was conducted to investigate the effects of iron (Fe, 40, 60, and 80 mg/kg) and Bacillus subtilis (2.5 × 10 9 and 5.0 × 10 9  CFU/kg) supplementation on reproductive performance, egg quality, nutrient digestibility, hormone levels, antioxidant indices, and hematological parameters in breeder geese. A total of one hundredtwenty 46-week-old Wulong breeder geese were randomly assigned to 1 of 6 dietary treatments with 4 replicates per treatment and 5 geese per replicate for 10 wk following 1 wk of adaption. Dietary Fe supplementation increased egg weight (P = 0.036), fertility (P = 0.022), serum total antioxidant capacity (P = 0.022), red blood cell (P = 0.001), hematocrit (HCT, P < 0.001), hemoglobin (HGB, P = 0.005), and mean corpuscular volume (MCV, P < 0.001). Dietary B. subtilis supplementation increased egg production (P = 0.025), eggshell thickness (P = 0.020), apparent phosphorus digestibility (P < 0.001), serum follicle stimulating hormone (P = 0.043), total antioxidant capacity (P < 0.001), HCT (P < 0.001), HGB (P < 0.001), and MCV (P = 0.025), and reduced malondialdehyde level (P = 0.008). The birds fed diets supplemented with 60 mg/kg Fe and 5 × 10 9  CFU/kg B. subtilis showed the highest percentage of hatched eggs (P = 0.004) and mean corpuscular hemoglobin (P < 0.001) among the 6 groups. Supplementation of 40 and 60 mg/kg Fe significantly increased the apparent digestibility of calcium compared with that of 80 mg/kg Fe in the birds fed 5.0 × 10 9  CFU/kg B. subtilis (P = 0.004). Supplementation with 60 and 80 mg/kg Fe in the birds fed 5 × 10 9  CFU/kg B. subtilis significantly decreased serum urea nitrogen level compared with other 4 groups (P = 0.022). In conclusion, the combination of Fe and B. subtilis effectively improves reproductive performance, eggshell quality, nutrient digestibility, antioxidant status, and hematopoietic function of breeder geese. Dietary addition of 60 mg/kg Fe and 5.0 × 10 9  CFU/kg B. subtilis was an optimum supplementation dose., (Copyright © 2020. Published by Elsevier Inc.)

Seipin, an evolutionary conserved protein, plays pivotal roles during lipid droplet (LD) biogenesis and is associated with various human diseases with unclear mechanisms. Here, we analyzed Caenorhabditis elegans mutants deleted of the sole SEIPIN gene, seip-1 Homozygous seip-1 mutants displayed penetrant embryonic lethality, which is caused by the disruption of the lipid-rich permeability barrier, the innermost layer of the C. elegans embryonic eggshell. In C. elegans oocytes and embryos, SEIP-1 is associated with LDs and is crucial for controlling LD size and lipid homeostasis. The seip-1 deletion mutants reduced the ratio of polyunsaturated fatty acids (PUFAs) in their embryonic fatty acid pool. Interestingly, dietary supplementation of selected n-6 PUFAs rescued the embryonic lethality and defective permeability barrier. Accordingly, we propose that SEIP-1 may maternally regulate LD biogenesis and lipid homeostasis to orchestrate the formation of the permeability barrier for eggshell synthesis during embryogenesis. A lipodystrophy allele of seip-1 resulted in embryonic lethality as well and could be rescued by PUFA supplementation. These experiments support a great potential for using C. elegans to model SEIPIN-associated human diseases., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2020. Published by The Company of Biologists Ltd.)

The study was conducted to evaluate the effects of dietary yeast β-glucan (YG) on performance and immune functions in breeder hens in a non-challenged setting. A total of 512 43-week-old Hy-Line Brown breeder hens were assigned into four treatments, and fed a basal diet with YG at 0, 50, 100 and 200 mg /kg for 8 weeks, respectively. Results showed that supplementation of YG did not affect production performance, but linearly increased hatchability (P < 0.05). Compared with the control, hens fed with 200 mg/kg YG had improved eggshell color and reduced mortality. Moreover, feeding 200 mg/kg YG significantly (P < 0.05) enhanced lymphocyte proliferation response to LPS, increased the percentage of peripheral blood CD3 + T cells and phytohemagglutinin (PHA) skin response; remarkably down-regulated splenic TLR4, IL-6 and TGF-β mRNA levels while upregulated TLR6 and IFN-γ mRNA levels (P < 0.05). In addition, inclusion of YG at 200 mg/kg considerably promoted the production of serum cytokines, total IgA, and specific antibody titers against BSA, AIV and NDV vaccine (P < 0.05). These results suggested that dietary inclusion of 200 mg/kg YG could improve eggshell color and fertile eggs hatchability and enhance cellular and humoral immune function of breeder hens in a nonchallenged setting without disturbing immune homeostasis., Competing Interests: Declaration of competing interest The authors declare that there are no conflicts of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The presence of extended-spectrum beta-lactamase (ESBL) producing Escherichia coli on poultry products is an important issue for veterinary and human health due to the zoonotic infection risk for producers and consumers. The present study focuses on testing the efficacy of six different disinfection methods on eggshell samples, aiming to reduce ESBL producing E. coli contamination on the hatching egg. Sterile eggshell cutouts were artificially contaminated with 108 cfu/ml CTX-M-1 producing E. coli and used as a carrier model to analyze the efficacy of six disinfection methods. The contaminated samples were separated into two groups; 1) contaminated and disinfected, 2) contaminated and non-disinfected. Six independent disinfection protocols were performed following product specifications and protocols. Each eggshell sample was separately crushed, and the total viable bacterial count was calculated to determine the disinfection efficacy. Five out of six tested methods (formaldehyde gassing, hydrogen peroxide + alcohol spray, essential oils spray, peracetic acid foam, and low energetic electron radiation) demonstrated a reduction or completely eliminated the initial ESBL producing E. coli contamination. One method (essential oils as cold fog) only partly reached the expected efficacy threshold (reduction of >102 cfu/ml) and the result differed significantly when compared to the reference method i.e. formaldehyde gassing., Competing Interests: The authors have declared that no competing interests exist.

The impact of feeding sources of n-3 fatty acids (FA) to ISA brown and Shaver white breeders and their female offspring during rearing on egg production, eggshell, tibia, and keel bone attributes was examined. Breeders were fed Control (CON) or CON + 1% dried microalgae (DMA: Aurantiochytrium limacinum) as the source of docosahexaenoic acid or CON + 2.6% of a coextruded mixture of full-fat flaxseed (FFF) and pulses as a source of α-linolenic acid. Day-old offspring were fed 1) breeder CON-pullet CON (CON-CON), 2) breeder CON-pullet DMA (CON-DMA), 3) breeder CON-pullet FFF (CON-FFF), 4) breeder DMA-pullet CON (DMA-CON), 5) breeder DMA-pullet DMA (DMA-DMA), 6) breeder FFF-pullet CON (FFF-CON), and 7) breeder FFF-pullet FFF (FFF-FFF). At 18 wk of age (WOA), pullets were fed a common layer diet to 42 WOA for egg production and bone quality assessments. There was no (P > 0.05) interaction between strains and diets and the main effect of diets on egg production, egg mass, and eggshell quality. There was an interaction (P = 0.008) between strain and diet on egg weight (EW); however, the strain effect on EW (P < 0.001) was such that ISA brown had heavier eggs than Shaver white. Shaver white had higher (P < 0.001) eggshell %, eggshell, and tibia breaking strength (BS), as well as tibia ash concentration compared with ISA brown hens. In contrast, ISA brown hens exhibited heavier (P < 0.05) tibia and keel bones. Feeding breeders DMA and pullets both sources of n-3 FA increased tibia medullary ash concentration compared with other diets (P < 0.001). Shaver white hens showed greater decline in tibia BS (83.7 vs. 96.3%) and ash content (84.1 vs. 94.3%) than ISA brown hens from 18 to 42 WOA (P < 0.05). Strain and diets exhibited independent effects on eggshell, tibia, and keel attributes. Provision of α-linolenic acid and docosahexaenoic acid to breeders and offspring improved tibia medullary ash concentration at 42 WOA., (Copyright © 2020. Published by Elsevier Inc.)

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 (17β-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., (Copyright © 2020. Published by Elsevier Inc.)

In order to investigate the effect of dietary Zn levels on laying performance, eggshell quality, and eggshell microstructure in Muscovy duck breeders under furnished cages. Firstly, the effects of age (35 weeks vs 40 weeks) and rearing system (littered floor vs furnished cage) on eggshell quality of laying duck breeders were studied (Exp. 1). Then, a total of 324 30-week-old Muscovy duck breeders were allotted into 3 dietary Zn groups with 6 replicates (18 ducks per replicate), including 0 mg Zn/kg (control-Zn group, C-Zn), 40 mg Zn/kg (normal-Zn group, N-Zn), and 140 mg Zn/kg (high-Zn group, H-Zn). The experimental period for 6 weeks was divided into 3 periods of 30-32, 32-34, and 34-36 weeks of age (Exp. 2). In Exp. 1, duck breeder eggs in the furnished cage system had lower the average shell thickness than birds in the littered floor system at 40 weeks of age (P < 0.05), not at 35 weeks of age. In Exp. 2, N-Zn and H-Zn groups had greater egg weight, egg production, and egg to feed ratio of duck breeders than C-Zn group (P < 0.05). Additionally, H-Zn group had higher laying rate, qualified egg ratio, and Haugh unit as well as lower mammillary cone width than C-Zn group (P < 0.05), with no differences between C-Zn and N-Zn groups (P > 0.05). Diet supplemented with 140 mg Zn/kg increased shell thickness and palisade layer thickness of duck breeders at 36 weeks of age (P < 0.05), but not at 32 and 34 weeks of age. In conclusion, diets with 40 or 140 mg Zn/kg improved egg production performance and egg quality of laying duck breeders during 30-36 weeks of age in a furnished cage system. Dietary supplementation of 140 mg Zn/kg level increased the ultrastructural palisade layer thickness contributing to greater eggshell thickness of duck breeders at 36 weeks of age.

The cuticle is the outmost layer of the eggshell and may affect the hatchability by modulating eggshell conductance. Three different solutions using acetic acid (AA), vinegar (V), and citric acid (CA) for cuticle removal by egg washing were developed, and the effects of cuticle removal on hatching performance of quail hatching eggs were evaluated. A total of 5,238 fresh quail hatching eggs were randomly divided into 9 treatments as follows: unwashed control, nondipped (CND); washed control, water dipped (CWD); standard control, 0.13% sodium hyperchlorite (CSH); 2% AA (AA2); 4% AA (AA4); 44.4% V (V2); 88.8% V (V4); 2% CA (CA2); and 4% CA (CA4). Overall, AA4, V4, and CA4 treatments significantly improved the hatchability of fertile eggs (95.42%, 94.16% and 95.66%, respectively) (P < 0.05) and the hatchability of CND, CWD, CSH, AA2, V2 and CA2 treatments were 90.98%, 93.00%, 92.27%, 79.44%, 90.37%, and 90.59%, respectively. The eggshell thickness and cuticle quality results showed that all AA, V, and CA solutions can effectively remove the quail eggshell cuticle, and AA4, V4 and CA4 significantly decreased eggshell thickness (P < 0.05). Microbial activity on the eggshell surface in all acid treatments was reduced significantly at day 0 of incubation (P < 0.05) and that significantly decreased than controlled treatments over the incubation period except AA2 treatment. Egg weight loss was lower for all acid treatments than that of the CND treatment (P < 0.05). There was no clear effect of treatments on chick quality. Hatch time in AA4, V4, and CA4 treatments slightly improved compared with controlled treatments (P > 0.05). There were no significant differences between treatments for chick livability and live weight at the first 21 D of life. Results of the present study indicate that cuticle removal with AA4, V4, or CA4 could effectively decrease the microbial activity on the eggshell surface during the incubation period and improve hatchability of quail hatching eggs without negative effects on hatch time and performance of quail chicks., (Copyright © 2020. Published by Elsevier Inc.)

This study was to investigate the effects of dietary zinc (Zn) supplementation on performance, zinc transporter gene expression, and immune function in aged laying hens. In experiment 1, twenty 31-week-old hens (young) and twenty 60-week-old hens (old) with the same genetic background were fed with the same diet for 4 weeks. In experiment 2, a basal diet supplemented with zinc sulfate (ZnS) and zinc glycine chelate (ZnG) at 30, 60, 90, and 120 mg Zn/kg to constitute nine experimental diets. Eight hundred and ten 60-week-old layers were distributed in a completely randomized experimental design with 9 treatments, 6 replicates of 15 birds each, and birds were fed for 10 weeks. In experiment 1, results showed that zinc and metallothionein (MT) concentration in the shell gland of old hens was significantly lower than young layers (P < 0.05). Zinc transporters ZnT1, 4, 5, 6, and 7 messenger RNA (mRNA) abundance in old layers were significantly lower versus the young (P < 0.05). In experiment 2, results indicated that dietary zinc supplementation did not significantly affect the laying rate, average feed intake, egg weight, feed conversion efficiency, broken egg rate, or mortality (P > 0.05). Supplemental ZnG significantly improved eggshell breaking strength than ZnS, with a higher alkaline phosphatase (ALP) activity and more abundant ZnT4 expression in shell gland versus ZnS (P < 0.05). ZnG supplementation at 90 mg Zn/kg affected the duodenal mucus by significantly increasing ZnT1, 6, 7, ZIP13, and MT-4 mRNA level (P < 0.05). Zinc level significantly increased bovine serum albumin (BSA) antibody concentration on 14 day after BSA injection (P < 0.05). Supplementation of ZnG improved eggshell quality of aged layers by upgrading zinc transporter expression in the shell gland and intestine also enhanced humoral immunity.

Neutral Electrolyzed Water (NEW) was tested in vitro and on artificially contaminated eggs against Salmonella enterica subsp. enterica or Escherichia coli. The antibacterial effect was measured 30 s after treatment. NEW microbicide activity results were compared against 2% citric acid and 0.9% saline solutions. NEW caused an in vitro decrease in Salmonella titers by ˃5.56 Log 10 CFU mL -1 and in artificially contaminated eggs by ˃1.45 Log 10 CFU/egg. When it was tested against E. coli, it decreased in vitro bacterial titers by ˃3.28 Log 10 CFU mL -1 and on artificially contaminated eggs by ˃6.39 Log 10 CFU/egg. The 2% citric acid solution caused an in vitro decrease of 0.4 Log 10 CFU mL -1 of Salmonella and E. coli and on eggs artificially contaminated with E. coli or Salmonella there was a decrease of 0.06 and 0.62 Log 10 CFU/egg respectively. We evaluated egg cuticle integrity by scanning electron microscopy after treatments with evaluated solutions; the 2% citric acid solution caused damage to the cuticle and exposed eggshell pores and no interaction of NEW or NaCl with the cuticle was observed. NEW treatment showed a fast-bactericidal effect in vitro and table eggs., Competing Interests: Declaration of competing interest Sandra Martinez-Vidal works at Esteripharma S.A. de C.V., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Effects of dietary available phosphorus (aP) and Ca levels and an Escherichia coli 6-phytase supplementation were studied in Lohmann LSL-Lite hens from 25 to 37 wk of age. Eighty-four hens were used in a completely randomized design with 7 treatments. The treatments were a positive control (PC) diet with 0.45% aP, 3.70% Ca, and 0.16% Na from 25 to 28 wk and 0.38% aP, 3.73% Ca, and 0.15% Na from 29 to 37 wk; a negative control (NC) diet, similar to the PC diet, with 0.22% aP, 3.00% Ca, and 0.13% Na from 25 to 28 wk and 0.19% aP, 3.02% Ca, and 0.13% Na from 29 to 37 wk; the NC diets supplemented with phytase at 150 (NC + 150), 300 (NC + 300), 600 (NC + 600), or 1,200 (NC + 1,200) phytase unit (FTU)/kg; and the PC diet supplemented with phytase at 1,200 (PC + 1,200) FTU/kg. Hen performance, eggshell, and bone quality were measured on a 4-wk basis. Bone breaking strength and ash and apparent ileal digestibility (AID) of P and Ca were determined at 37 wk. One- and 2-way ANOVA were conducted, and Tukey's range test was used to compare multiple means where P ≤ 0.05. No differences in hen performance, eggshell quality, bone breaking strength, bone ash, and P digestibility were observed between the PC and the NC treatments. The NC hens had lower cortical (P < 0.001) and trabecular + medullary bone mineral density (P = 0.004) and total bone mineral content (P < 0.001) than the PC hens. The PC + 1,200 increased cortical bone mineral density (P < 0.001). The reductions of aP and Ca in the NC diet were not deficient for performance but had a minor impact on bone mineralization. The NC + 600 and NC + 1,200 increased AID of P (P = 0.024), and all phytase treatments except the NC + 150 increased AID of Ca (P = 0.010) compared with the NC diet., (Copyright © 2020. Published by Elsevier Inc.)

The objective of this study was to investigate the effects of low inclusion levels of organic trace minerals (iron, copper, manganese, and zinc) on performance, eggshell quality, serum hormone levels, and enzyme activities of laying hens during the late laying period. A total of 405 healthy hens (HY-Line White, 50-week-old) were randomly divided into 3 treatments, with 9 replications per treatment and 15 birds per replication. The dietary treatments included a basal diet supplemented with inorganic trace minerals at commercial levels (CON), a basal diet supplemented with inorganic trace minerals at 1/3 commercial levels (ITM), and a basal diet supplemented with proteinated trace minerals at 1/3 commercial levels (TRT). The trial lasted 56 D (8 wk). Compared with the CON group, the ITM group showed decrease in (P < 0.05) egg production, eggshell strength, eggshell palisade layer, palisade layer ratio, serum estrogen, luteinizing hormone, glycosaminoglycan concentration, and carbonic anhydrase activity and increase in (P < 0.05) egg loss and mammillary layer ratio. However, the TRT group almost kept all the indices close to the CON group (P > 0.05). Furthermore, hens fed with low inclusion levels of organic trace minerals had smaller mammillary knobs (P < 0.05) than those in the CON and ITM groups. In conclusion, hens fed with low inclusion levels of proteinated trace minerals had better performance and eggshell strength than those fed with identical levels of inorganic compounds; organic trace minerals improved eggshell quality by improving the eggshell ultrastructure of laying hens during the late laying period., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

The spatial network structure of Dergall is based on substances nontoxic to humans and the environment which, when applied on solid surfaces, creates a coating that reduces bacterial cell adhesion. The bacteriostatic properties of siloxanes are based on a purely physical action mechanism which excludes development of drug-resistant microorganisms. The aims of the present study were to 1) evaluate a Dergall layer formed on the eggshell surface regarding the potential harmful effects on the chick embryo; 2) evaluate antimicrobial activity and estimate the prolongation time of Dergall's potential antimicrobial activity. Dergall at a concentration of 0.6% formed a layer on the eggshell surface. In vitro testing of the potential harmful effects of Dergall by means of a hen embryo test of the chorioallantoic membrane showed no irritation reaction at a concentration of 3% and lower. The hatchability of the groups sprayed with a Dergall water solution with a concentration of 0 to 5% was 89.1 to 93.8% for fertilized eggs (P > 0.05) but decreased to 63.7% (P < 0.05) in the group sprayed with a 6% concentration of the solution. This phenomenon was caused by embryo mortality in the first week of incubation. At the concentration of 0.6%, Dergall exhibited strong antibacterial properties against bacteria such as Staphylococcus aureus, Escherichia coli, Shigella dysenteriae, Shigella flexneri, and Salmonella typhimurium. For Streptococcus pyogenes, the highest antibacterial activity of Dergall was reported in the concentrations of 100 and 50%. For Pseudomonas aeruginosa, no antibacterial activity of Dergall was generally observed, but in vivo testing showed a strong decrease of all gram-negative bacteria growth. Moreover, a prolonged antimicrobial effect lasting until 3 D after disinfection was observed, which makes Dergall a safe and efficient disinfectant., (Copyright © 2019. Published by Elsevier Inc.)

This study demonstrates the effects of progesterone on eggshell quality and ultrastructure by injecting progesterone into laying hens 2 and 5 h post-oviposition, respectively. Progesterone injected 2 h post-oviposition (P 4 -2 h) improved eggshell quality with a significant decrease (P < 0.01) in the thickness of the mammillary layer and a significant increase (P < 0.01) in the thickness of the effective layer in the eggshell ultrastructure compared to the control. Progesterone injected 5 h post-oviposition (P 4 -5 h) damaged the eggshell quality by significantly reducing (P < 0.01) the effective layer thickness. Progesterone injected delayed obviously (P < 0.01) the following oviposition. Moreover, the concentrations of Thr, Cys, Leu, Lys, and His in the eggshell membranes were significantly higher (P < 0.05) in the P 4 -2 h treated hens whereas Val and Lys were significantly lower (P < 0.05) in P 4 -5 h treated hens compared to the control. Therefore, progesterone shows paradoxical effects on eggshell quality depending on the injection time-points post-oviposition, which could explain the contradictions in previous related reports. P 4 injected affected the content of amino acids in eggshell membranes, especially lysine which contributed to eggshell quality. In addition, P 4 injected 2 h after oviposition improved eggshell quality by promoting the premature fusion of mammillary knobs. This work contributed to a novel insight to understanding the mechanism of improving eggshell quality., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2019 Elsevier Inc. All rights reserved.)

The objective of this study was to evaluate the effect of dietary chloride (Cl) levels on performance, eggshell mechanical quality, and ultrastructure in layers based on the substitution of sodium chloride (NaCl) by sodium sulfate (Na 2 SO 4 ). Three hundred sixty Jing Brown laying hens aged 43 wk were randomly divided into 5 groups and fed with corn-soybean meal diets containing 0.06, 0.10, 0.15, 0.20, and 0.25% total Cl inclusion. Every group had 8 replicates of 9 birds each. The feeding trial lasted for 12 wk. The results showed dietary 0.06% Cl due to complete substitution of NaCl by Na 2 SO 4 , depressed performance (P < 0.05) from 45 to 54 wk of age, increased serum creatinine level (P < 0.05), and caused visible renal tubular atrophy. Dietary Cl levels quadratically affected breaking strength, thickness, and weight of eggshell (P < 0.05). Better eggshell quality could be obtained when NaCl was partly replaced by Na 2 SO 4 in laying hen diets maintaining Cl level at 0.10 or 0.15%. Moreover, the eggshell ash content was affected by Cl levels in a quadratic (P < 0.001) manner, with higher values observed in the 0.10 and 0.15% Cl groups (P < 0.05). Besides, the eggshell ultrastructural data showed that the total thickness and effective thickness significantly increased (P < 0.05) and mammillary thickness decreased (P < 0.05) in the group of dietary 0.15% Cl compared with the groups of 0.06 and 0.25% Cl. In conclusion, the complete substitution of dietary NaCl by Na 2 SO 4 may induce Cl deficiency and depress laying performance and eggshell quality. Na 2 SO 4 could partly replace NaCl in diets for laying hens (43-54 wk of age) without adverse effects on performance at the dietary Cl level from 0.10 to 0.25%. Better eggshell quality could be obtained when NaCl was partly replaced by Na 2 SO 4 in laying hen diets maintaining Cl level at ∼0.15%., (Copyright © 2019. Published by Elsevier Inc.)

Reproductive disorders in birds are the most characteristic effects of DDT contamination of wildlife. Experimental exposure of avian eggs to the estrogenic substance o,p'-DDT causes abnormal development of the reproductive tract (shortening of the left oviduct and aberrant development of the right oviduct) and eggshell thinning in mature birds, but it is still not known how eggshell thinning occurs in the abnormal oviduct. To fill this information gap, we examined the histology of the uterine part of the oviduct in Japanese quail treated in ovo with o,p'-DDT or a synthetic estrogen, diethylstilbestrol (DES), and we performed immunohistochemical staining for the calcium-binding proteins CALB1, SPP1, and TRPV6. Both o,p'-DDT-treated and DES-treated quail had few, and scattered, gland cells in the left uterus, unlike vehicle controls, in which gland cells tightly occupied the lamina propria. The aberrantly developed right uterus retained all the components of the normal left uterus, but in immature form. Immunostaining for CALB1, SPP1, and TRPV6 was greatly reduced by both o,p'-DDT and DES; SPP1 and TRPV6 immunostaining patterns, in particular, differed distinctly from those in the controls. These findings suggest that CALB1, SPP1, and TRPV6 are molecular factors, decreased production of which is responsible for eggshell thinning. Our findings also could contribute to understanding of the eggshell formation mechanism in birds.

The objective of this experiment was to investigate the effects of various dietary concentrations of vitamin D3 (D3) on pullet and laying hen performance, eggshell quality, bone health, and yolk D3 content from day of hatch until 68 wk of age. Initially, 440 Hy-line W36-day-old chicks were randomly assigned to 5 dietary treatments: 1,681 (control); 8,348; 18,348; 35,014; 68,348 IU D3/kg. At 17 wk of age, pullets were assigned to experimental diets with 12 replicate groups of 6 birds. At 17 wk of age, pullets fed diets containing 8,348 and 35,014 IU D3/kg had an increased bone mineral density in comparison to the control fed birds (P ≤ 0.01). Body weights of pullets fed the diet with 68,348 IU D3/kg were lower than other treatments (P ≤ 0.01). Hen-housed egg production (HHEP) of hens fed the 35,014 IU D3/kg diet was increased in comparison to control-fed hens (P ≤ 0.01), whereas HHEP of those fed 68,348 IU D3/kg diet was reduced in comparison to all other treatments (P ≤ 0.01). Shell breaking strength of eggs from hens fed 8,348, 35,014 and 68,348 IU D3/kg was increased in comparison to eggs from control-fed birds (P ≤ 0.01). Fat-free tibia ash content of hens fed any of the diets supplemented with D3 (8,348 to 68,348 IU D3/kg) was increased in comparison to control-fed hens (P ≤ 0.05). Yolk D3 content increased linearly with dietary D3 and the D3 transfer efficiency for the control, 8,348 IU, 18,348 IU, 35,014 IU, and 68,348 IU D3 treatments were 8.24, 10.29, 11.27, 12.42, and 12.06%, respectively. These data suggest that supplementation of dietary D3 up to 35,014 IU D3/kg feed maintained if not increased laying hen performance and enhanced pullet and laying hen skeletal quality as well as yolk D3 content and eggshell quality. Feeding pullets at a higher level 68,348 IU of D3 resulted in reduced growth and ultimately decreased performance of laying hens., (© 2019 Poultry Science Association Inc.)

Salmonella enteritidis (S. enteritidis) infection has been recognized as one of the most common bacterial causes of human gastroenteritis worldwide and is closely associated with eggs. Slightly acidic electrolyzed water (SAEW) is an emerging environmentally friendly technology for disinfecting eggshell surfaces to remove dirt and pathogenic microorganisms. However, the efficiency of SAEW could be affected by the presence of manure. UV-based advanced oxidation processes have been studied to improve the microorganism's inactivation effect of disinfection. Therefore, in this study, the synergistic bactericidal efficacy of SAEW and UV-C light (ultraviolet lamp, λ = 254 nm) for inactivation of S. enteritidis on artificially inoculated eggshells with or without manure was evaluated, and the bactericidal efficacy of different combination treatments of SAEW and UV-C light was compared. Without manure interference, complete inactivation (reduction of 6.54 log10 CFU/g) of S. enteritidis on the surface of eggshells was achieved following a 4-min treatment with SAEW+UV at an available chlorine concentration (ACC) of 20 mg/L. In the presence of manure, a 3.02 log reduction was achieved following a 4-min treatment with SAEW+UV at an ACC of 30 mg/L. Simultaneous treatment with SAEW and UV light exhibits higher bactericidal activity for eggshells than other combination process methods with UV and SAEW. The results suggest that the combined treatment of SAEW+UV is a novel method to enhance the microbial safety of eggshells., (© 2019 Poultry Science Association Inc.)

Adequate dietary Ca and available phosphorus (avP) are essential to long-term egg production and bone health in laying hens. The effects of dietary Ca and avP levels and Buttiauxella sp. phytase (BSP) were studied in Lohmann LSL Lite hens from 30 to 70 wk of age (woa). Hens (n = 456; 4 per cage) were fed either a primary breeder recommendation-based diet (positive control; PC); the PC with avP and Ca levels reduced by 0.146 and 0.134% of the diet, respectively, without (NC) or with 300 FTU/kg BSP (NC+BSP). Egg production, BW, feed intake, FCR, and eggshell quality from 30 to 70 woa, and apparent ileal digestibility of P (AIDP) and Ca (AIDCa), and bone quality at 32, 48, and 70 woa were measured. The avP and Ca levels in the NC diet were not clinically deficient, as most parameters were unaffected by diet. Hen BW from 34 to 70 woa tended to be 2.9% greater (P = 0.076) for PC and NC+BSP compared to NC. Mid-diaphysis cortical bone mineral content (CBMC) tended to be 10% and 9% higher (P = 0.065) in the NC+BSP hens than in NC hens at 48 and 70 woa, respectively. AIDP of NC+BSP was 24% greater (P = 0.034) than of NC at 32 woa and tended to be 18% greater (P = 0.082) than AIDP of PC at 48 woa, and 25% lower than of NC and PC at 70 woa (P = 0.028). AIDCa was 25% lower for NC+BSP than PC at 48 woa only (P = 0.037). The avP and Ca sufficiency in the NC diet limited the opportunity to determine a phytase effect. Although the supplemental BSP tended to increase BW and 48 and 70 woa CBMC, and increased 32 woa AIDP, the efficacy of BSP could not be determined due to the lack of an NC effect on most parameters. Commercial laying hens can maintain health and productivity at lower than recommended levels of dietary Ca and avP; phytase supplementation may allow for even further reductions., (© 2019 Poultry Science Association Inc.)

Background: The objective of this study was to assess the effect of locally administered ibuprofen (IBU) on transected peripheral nerve regeneration and functional recovery., Methods: Seventy-five male Wistar rats were divided into five experimental groups (N.=15), randomly: In authograft group (AUTO) a segment of sciatic nerve was transected and reimplanted reversely. In transected group (TC), left sciatic nerve was transected and stumps were fixed in the adjacent muscle. In treatment group defect was bridged using an egg shell membrane conduit (ESM/IBU) filled with 10 µL ibuprofen (100 ng/mL). In ESM conduit group (ESM), the conduit was filled with phosphate-buffered saline alone. In sham-operated group (SHAM), sciatic nerve was exposed and manipulated. Each group was subdivided into three subgroups of five animals each and regenerated nerve fibers were studied 4, 8 and 12 weeks after surgery., Results: Behavioral testing, biomechanical studies, sciatic nerve functional study, electrophysiological, gastrocnemius muscle mass and morphometric indices confirmed faster recovery of regenerated axons in ESM/IBU than ESM group (P<0.05). In immunohistochemistry, location of reactions to S-100 in ESM/IBU was clearly more positive than that in ESM group., Conclusions: Ibuprofen accelerated and improved functional recovery and morphometric indices of sciatic nerve. This study is expected to set a stage for testing the ibuprofen in the human patients.

1. In this study, 240 ISA Brown hens were fed diets containing different levels of hybrid rye, and the influence of xylanase addition on laying performance and egg quality was evaluated. 2. Birds were allocated to 10 treatment groups with 12 replicates (cages) of two hens and were fed, from week 26 to 50, isocaloric and isonitrogenous experimental diets. A 5 × 2 experimental arrangement was applied, using diets with increasing level of rye (0%, 10%, 15%, 20% or 25%) with or without xylanase supplementation (200 mg/kg of feed; Ronozyme WX (CT) with minimum xylanase activity of 1,000 FXU/g). 3. Increasing dietary level of rye did not affect daily mass of eggs, mean egg weight or feed conversion ratio ( P > 0.05). Laying rate decreased in all groups fed with rye. Egg and eggshell quality indices were unaffected by dietary rye grain ( P > 0.05); however, rye inclusion significantly decreased yolk colour on the DSM scale ( P < 0.05). In comparison with the control group, high dietary levels of rye (25%) significantly increased viscosity of small intestine content ( P < 0.05). Diet supplementation with xylanase had no significant effect on egg production indices and egg quality (except for yolk colour) but decreased the viscosity of intestinal content in laying hens fed high levels of rye ( P < 0.05). 4. The results of this experiment suggest that rye may be incorporated to a level of 25% in the diet of laying hens without any strong negative effect on egg performance, while xylanase added to high-rye grain reduced the viscosity of intestinal content; however, it did not positively affect the laying performance or egg quality.

Designing reasonably priced bioactive organic-inorganic hybrid materials featuring osteogenic activity has received a significant interest in the bone tissue engineering field. In this study, eggshell membrane (ESM), which consists mostly of proteins exhibiting fibrous network structure, was utilised innovatively as a template to prepare eggshell membrane/hydroxyapatite (ESM-HA) composites using a versatile biomimetic mineralisation technique. In addition, the surface morphology and composition, hydrophilicity, crystallinity, and thermal stability of both sides of ESM and ESM-HA composites were systematically studied. Results indicated that both sides of ESM exhibited excellent biomimetic mineralisation ability, and the hydrophilicity and thermal stability of ESM were effectively improved by the introduction of HA. Moreover, in vitro experiments on MC3T3-E1 cells revealed that the inner side of the ESM was more beneficial to cell proliferation and adhesion than the outer side. Remarkably, the proliferation, adhesion and spreading, as well as the alkaline phosphatase (ALP) activity and expression of bone-related genes and proteins (runt-related transcription factor 2, ALP, collagen type I, and osteocalcin) on both sides of ESM-HA composites were significantly higher compared to those of the original ESM. These findings suggested that ESM-HA composites obtained using biomimetic mineralisation could be potential new materials for future bone tissue repair., (Copyright © 2019 Elsevier B.V. All rights reserved.)

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., (© 2019 Poultry Science Association Inc.)

The present work was carried out to investigate the effects of dietary propolis supplementation to laying Japanese quail (Coturnix coturnix japonica) on egg production, egg quality, physiological and immunological aspects under heat stress conditions. A total of 200, 21-day-old, Japanese quail females were distributed equally into standard wired cages in two identical environmentally-controlled rooms (10 cages per room, 10 birds per cage). From 29-70 d of age, the quail birds in the first room remained at a normal temperature of 24°C (C group), whereas the quail birds in the second room were kept under heat stress at 35°C (HS group). Each group was further assigned to 2 propolis subgroups (5 cages per subgroup); one of them received a basal diet without propolis supplementation (-PR subgroup), while, the other received 1 g propolis/ kg basal diet (+PR subgroup). In the present study, performance and egg production of laying quail were significantly (P<0.001) impaired by HS treatment and improved by the PR treatment. Similarly, the negative and positive effects of HS and PR, respectively, were appeared on the egg shell thickness and yolk index. Stress indicators in laying quail were significantly (P<0.001) increased by HS, while, PR significantly (P<0.05) moderated these levels in the HS+PR group when compared to the HS-PR quail group. In addition to the positive impact of PR on the plasma levels of calcium, phosphorus, and albumin, it also normalized the plasma levels of alanine aminotransferase and cholesterol in the heat-stressed quail birds. Moreover, the quail birds in the HS groups expressed lower immunological aspects than those in the C group, while, the addition of propolis to the diets enhanced the immune status of laying quail birds under HS conditions. These results strongly suggest that dietary propolis supplementation could be a successful attempt to maintain the performance and egg production of laying Japanese quail at convenient levels under heat stress conditions., Competing Interests: The authors have declared that no competing interests exist.

One hundred and twenty Cobb 500 hens, 20 wk of age, were randomly allocated into individual cages with the objective of estimating their Zn requirements. The study was composed of 3 phases: adaptation to cages (basal diet), depletion (deficient diet containing 18.7 ± 0.47 ppm Zn) for 7 wk, and experimental phases. Hens were fed diets with graded increments of Zn sulfate heptahydrate (ZnSO4·7H2O), totaling 18.7 ± 0.47, 50.3 ± 10.6, 77.3.0 ± 11.0, 110.2 ± 12.8, 140 ± 12.2, and 170.6 ± 13.2 ppm analyzed Zn in feeds for 12 wk (experimental phase). Requirements of Zn were done using quadratic polynomial (QP), broken line quadratic (BLQ), and exponential asymptotic (EA) models. In general, the non-linear statistical models were the ones that best fit the results in this study. Requirements obtained for hen day egg production and settable egg production were 83.3, 78.6 ppm and 61.4, 65.4 ppm for period of 33 to 36 wk, and 63.3, 53.1 and 60.4, 46.1 ppm for period of 37 to 40 wk, and 62.8, 52.8, and 67.7, 62.1 ppm for period of 41 to 44 wk, respectively, using BLQ and EA models. Total eggs and total settable eggs produced per hen had Zn requirements estimated as 75.7, 64.7 ppm, and 56.5, 41.5 ppm, respectively, for BLQ and EA models, whereas for alkaline phosphatase and eggshell percentage were 161.8, 124.9 ppm and 126.1, 122.4 ppm, using QP and BLQ models. Maximum responses for Zn in yolk for periods of 37 to 40 and 41 to 44 wk were 71.0, 78.1 and 64.5, 59.6 ppm, respectively, using BLQ and EA models. Breaking strength had Zn requirements estimated at 68.0 and 96.7 ppm, whereas eggshell palisade layer and eggshell thickness were maximized with 67.9, 67.9 ppm, and 67.7, 64.4 ppm, respectively, for BLQ and EA models. The average of all Zn requirement estimates obtained by EA and BLQ models in the present study was 72.28 ppm or 11.1 mg/hen/d., (© 2018 Poultry Science Association Inc.)

Calcium and vitamin D are critical in attenuating metabolic Ca insufficiency often observed in older hens. We investigated interactive effects of dietary Ca and top-dressed 25-hydroxy vitamin D3 (25OHD3) on egg production, eggshell quality (thickness, EST, and breaking strength, ESBS), serum Ca, kidney, liver, and bones attributes in 74-wk old Lohmann LSL-lite layers. A total of 4 levels of Ca (3.0, 3.5, 4.0, and 4.5%) and 3 levels of 25OHD3 (0, 69, and 138 μg/kg) were tested. All diets had basal level of 3,300 IU of vitamin D3/kg, were allocated to 84 individually housed hens (n = 7), and fed to 81 wk of age. Dual-energy X-ray absorptiometry was used to measure bone mineral content (BMC) and density (BMD) in left ulna, femur, and tibia at week 74 (13 spare birds, baseline) and week 81. Birds fed diets containing 3.0, 3.5, 4.0, and 4.5% Ca consumed 3.4, 4.1, 4.2, and 4.8 Ca g/b/d. Interaction (P < 0.05) between Ca and 25OHD3 was such that 25OHD3 linearly increased egg weight and percentage of eggs graded as jumbo (≥70 g) at 3.0, 3.5, and 4.5% Ca levels. There was no interaction (P > 0.05) between Ca and 25OHD3 on the rest of the parameters. Calcium increased EST and ESBS quadratically (P < 0.05), EST increased with Ca intake up to 4.1 g of Ca/b/d whereas Ca intakes (3.4 and 4.8 g of Ca/b/d) had poor ESBS relative to intermediate Ca intakes. Kidney ash concentration increased linearly (P = 009) with Ca intake. However, BMC and BMD in ulna, tibia, and femur declined (P < 0.05) at week 81 relative to baseline. In conclusion, top dressed 25OHD3 linearly increased egg weight at various Ca intakes with no negative effect on eggshell quality or bone attributes. Feeding 4.1 to 4.2 g of Ca/b/d improved eggshell quality, however, higher intake deteriorated eggshell quality and increased renal ash accumulation with no benefit on skeletal integrity., (© 2018 Poultry Science Association Inc.)

In order to evaluate the effect of rosemary oil and micro mineral sources on the performance and egg quality of laying hens, 288 hens were used and distributed in a completely randomized design using a 2x3 factorial (mineral sources x rosemary oil) with six different diets and six replications. Diets were formulated containing inorganic or organic minerals, with or without added rosemary oil (100 and 200 mg kg-1). The treatments showed interaction for average egg weight, yolk color, albumen and yolk percentage, with significant results from the use of organic minerals compared to inorganic minerals. The addition of 200 mg kg-1 rosemary oil in diets improved laying rate, egg mass, specific gravity, number of pores on the shell when compared to other treatments. In conclusion, the use of 200 mg kg-1 of rosemary oil improves the performance of red laying hen eggs. The association between organic minerals and 100 mg kg-1 rosemary oil in laying hen diets increases yolk color and percentage of albumen. The use of organic minerals is superior to inorganic minerals as to improve the quality of eggs, increasing average egg weight, yolk color and percentage of albumen.

1. An experiment was conducted to determine optimal non-phytate phosphorus (NPP) concentrations for White Leghorn (WL) layers (22-72 weeks) fed diet containing 38 g Ca/kg. 2. Eight diets with graded concentrations (1.5-3.25 g/kg in increments of 0.25 g) of NPP were prepared. Each diet was fed to eight pen replicates containing 88 birds in each. Performance data was evaluated in three different phases (phase I-22-37 weeks, phase II-38-53 weeks and phase III-54-72 weeks). Optimum levels of NPP were determined by fitting a quadratic polynomial (QP) regression model. 3. Egg production (EP) was not affected (P = 0.059) by the concentration of NPP and interaction between NPP and diet phase was non-significant, indicating that the lowest concentration (1.5 g/kg diet) of NPP used in the study was adequate across the three phases. However, EP was influenced by phase (P < 0.001). 4. Optimum concentration of NPP for feed intake (FI) was estimated to be 1.5, 1.71 and 2.40 g/kg diet during phases I, II and III, respectively. FI per egg mass (EM) or feed efficiency (FE) responded quadratically with NPP and also differed significantly between phases. Optimum concentration of NPP for FE during phases I, II and III was 1.5, 2.56 and 2.32 g/kg diet, respectively. 5. Egg weight (EW), EM, shell weight and thickness were not affected by NPP concentration although all of these variables (except shell weight) were influenced by phases. 6. Breaking strength of tibia and Ca contents in tibia ash were not affected by the concentration of NPP, but bone ash and P contents in tibia ash were influenced (P < 0.001) by NPP. Predicted optimal concentrations of NPP for responses for tibia ash at 44 or 72 weeks, tibia ash P at 44 weeks and tibia ash P at 72 weeks were 1.55, 2.63 and 1.5 g/kg diet, respectively. 7. Based on the results, it was concluded that WL layers required 1.5 g, 2.63 g and 2.4 g, respectively/kg diet during phase I, II and III with the calculated daily intake of 137.3, 278.3 and 262 mg NPP/b/d.

The study aimed to determine the effects of methionine hydroxyl analog chelated zinc (MHA-Zn) on laying performance, eggshell quality and mineral deposits, and the activities of Zn-containing enzymes on aged laying hens. A total of 960 layers (Hy-Line Grey, 57 wk old) were fed a basal diet (Zn: 35.08 mg/kg) without extra zinc for 2 wk. During the ensuing 14 wk, birds were randomly divided into 4 groups according to body weight and laying rate, with 8 replicates per treatment, and each group had 8 replicates of 30 hens. Four levels of Zn (ZnSO4: 80 mg/kg; MHA-Zn: 20, 40, 80 mg/kg) were added to the diet, respectively. The results shown that dietary Zn did not affect laying rate, average egg weight, average daily feed intake, or feed conversion ratio (P > 0.05); however, compared to the inorganic group, dietary supplementation with 40 or 80 mg/kg MHA-Zn decreased broken egg rate significantly in the whole period (P < 0.05), while significantly increased eggshell weight in week 62 to 72, eggshell thickness and eggshell strength in wk 66 to 72, eggshell weight percent and eggshell density in week 62 to 72 (P < 0.05). Besides, dietary supplementation with different sources and levels of Zn did not affect ash concentration of eggshell (P > 0.05), whereas dietary supplementation with 80 mg/kg MHA-Zn improved the Zn and Ca concentrations of eggshells and carbonic anhydrase (CA) activity of liver, and 40 mg/kg MHA-Zn increased Zn concentration of liver (P < 0.05). Moreover, no significant differences in alkaline phosphatase activity were observed among the treatment groups (P > 0.05). Therefore, dietary supplementation with 40 mg/kg MHA-Zn can improve eggshell quality by promoting Ca deposition and CA activity.

The objective of this study was to determine the effects of mercury chloride (HgCl 2 ) on laying performance, egg quality, hepatic and renal histopathology, and serum biochemical profiles in laying hens. A total of 768 Hy-line brown laying hens aged 40 weeks were randomly allocated to four groups (8 pens per group and 24 hens per pen). The concentrations of mercury (Hg) in four groups were 0.280, 3.325, 9.415, and 27.240 mg/kg. Results revealed that dietary Hg could significantly reduce laying performance (P < 0.05) and egg quality (P < 0.05) and was dose-dependently deposited in albumen, yolk, eggshell, and whole egg. Meanwhile, the thicknesses of palisade layer, mammillary layer, and total layer, and the percent of palisade layer were significantly decreased (P < 0.05), while the percent of mammillary layer was sharply increased (P < 0.05) in eggshell. In addition, with increasing dietary dosage of Hg, accumulation of Hg in viscera was significantly increased (P < 0.05), and histopathological damages in liver and kidney were more and more severe. Serum alanine aminotransferase, aspartate aminotransferase, and globulin were significantly increased (P < 0.05), while serum albumin and albumin to globulin ratio were significantly decreased (P < 0.05) in 27.240 mg/kg Hg group. Blood urea nitrogen, uric acid, and creatinine were significantly increased (P < 0.05) in 3.325, 9.415, and 27.240 mg/kg Hg groups. These results suggested that dietary HgCl 2 could reduce laying performance and egg quality with hepatic and renal function disorders in laying hens.

It has been demonstrated that tea polyphenol (TP) epigallocatechin-3-gallate (EGCG) can confer protection against vanadium (V) toxicity in laying hens; however, our understanding of the molecular mechanisms beyond this effect are still limited. In this study, 360 hens were randomly assigned to the 3 groups to study whether the potential mechanism P38MAPK-Nrf2/HO-1 signaling pathway is involved in the protective effect of EGCG on eggshell pigmentation in vanadium challenged laying hens. Treatments included a control group, a 10 mg/kg V (V10), and a V10 plus 130 mg/kg of EGCG group (V10+EGCG130). Both eggshell color and protoporphyrin IX were decreased in the V10 group compared with the control diet, while EGCG130 treatment partially improved shell color and protoporphyrin IX (P < 0.05). The V10 exposure induced higher cell apoptosis rate and oxidative stress in birds as evidenced by the histological apoptosis status, decreased uterine glutathione-S transferase (GST) and high abundance of malondialdehyde (MDA) compared with the control group, whereas EGCG130 markedly alleviated oxidative stress via reducing MDA generation (P < 0.05). Dietary vanadium reduced ferrochelatase, NF-E2-related factor 2 (Nrf2), and heme oxygenase (HO-1) mRNA expression, while EGCG up-regulated Nrf2 and HO-1 expression (P < 0.05). Protein levels of Nrf2, HO-1 and phospho-p38 (P-P38) MAPK were reduced in V10 group, while dietary supplementation with 130 mg/kg EGCG markedly increased Nrf2, HO-1 and P-P38 MAPK protein levels in the uterus compared with the V10 group (P < 0.01). In conclusion, EGCG improved eggshell color and antioxidant system in V10-challenged hens, which seems to be associated with P38MAPK-Nrf2/HO-1 signaling pathway.

One-hundred-twenty Cobb 500 hens, 20 wk of age, were randomly allocated into individual cages with the objective of estimating Cu requirements. After being fed a Cu deficient diet for 4 wk, hens were fed diets with graded increments of supplemental Cu (0.0; 3.5; 7.0; 10.5; 14; and 17.5 ppm) from Cu sulfate (CuSO4 5H2O), totaling 2.67; 5.82; 9.38; 12.92; 16.83; and 20.19 ppm analyzed Cu in feeds for 20 weeks. Estimations of Cu requirements were done using exponential asymptotic (EA), broken line quadratic (BLQ), and quadratic polynomial (QP) models. Obtained Cu requirements for hen d egg production and total settable eggs per hen were 6.2, 7.3, and 12.9 ppm and 8.1, 9.0, and 13.4 ppm, respectively, using EA, BLQ, and QP models. The QP model was the only one having a fit for total eggs per hen with 13.1 ppm Cu as a requirement. Hemoglobin, hematocrit, and serum Cu from hens had requirements estimated as 13.9, 11.3, and 18.5, ppm; 14.6, 13.0, and 19.0 ppm; and 16.2, 14.6, and 14.2 ppm, respectively, for EA, BLQ, and QP models. Hatching chick hemoglobin was not affected by dietary Cu, whereas requirements estimated for hatching chick hematocrit and body weight and length were 10.2, 12.3, and 13.3 ppm using EA, BLQ, and QP models; and 6.8 and 7.1 ppm, and 12.9 and 13.9 ppm Cu using EA and BLQ models, respectively. Maximum responses for egg weight, yolk Cu content, and eggshell membrane thickness were 14.9, 12.7, and 15.1 ppm; 15.0, 16.3, and 15.7 ppm; and 7.3, 7.8, and 14.0 ppm Cu, respectively, for EA, BLQ, and QP models. Yolk and albumen percentage were adjusted only with the QP model and had requirements estimated at 11.0 ppm and 11.3 ppm, respectively, whereas eggshell mammillary layer was maximized with 10.6, 10.1, and 14.4 ppm Cu using EA, BLQ, and QP models, respectively. The average of all Cu requirement estimates obtained in the present study was 12.5 ppm Cu.