Coccidia vaccination is a common prac-tice in the poultry industry. However, research is lacking regarding the optimal nutritional support for coccidia vaccinated broilers. In this study, broilers were vacci-nated with coccidia oocyst at hatch and were fed with a common starter diet from 1 to 10 d. On d 11, the broilers were randomly assigned to groups in a 4 pound 2 factorial arrangement. Briefly, the broilers were fed one of four diets containing 0.6, 0.8, 0.9, and 1.0% of standardized ileal digestible methionine plus cysteine (SID M+C), respectively, from 11 to 21 d. On d 14, the broilers from each diet group were orally gavaged with either PBS (Mock challenge) or Eimeria oocysts. Compared to PBS-gavaged broilers and regardless of dietary SID M +C levels, the Eimeria-gavaged broilers had 1) decreased gain-to-feed ratio (15-21 d, P = 0.002; 11-21 d, P = 0.011); 2) increased fecal oocysts (P < 0.001); 3) increased plasma anti-Eimeria IgY (P = 0.033); and 4) increased intestinal luminal interleukin-10 (IL-10; duo- denum, P = 0.039; jejunum, P = 0.018) and gamma interferon (IFN-g; duodenum, P < 0.001; jejunum, P = 0.017). Regardless of Eimeria gavage, broilers fed 0.6% SID M+C had decreased (P<0.001) body weight gain (15-21 and 11-21 d) and gain-to-feed ratio (11-14, 15-21, and 11-21 d) when compared to those fed >= 0.8% SID M+C. Eimeria challenge increased (P < 0.001) duo-denum lesions when the broilers were fed with 0.6, 0.8, and 1.0% SID M+C, and increased (P = 0.014) mid -intestine lesions when the broilers were fed with 0.6 and 1.0% SID M+C. An interaction between the two experi-mental factors was detected on plasma anti-Eimeria IgY titers (P = 0.022), as coccidiosis challenge increased plasma anti-Eimeria IgY titers only when the broilers were fed with 0.9% SID M+C. In summary, the dietary SID M+C requirement for grower (11-21 d) broilers vaccinated with coccidiosis was ranged from 0.8 to 1.0% for optimal growth performance and intestinal immu-nity, regardless of coccidiosis challenge.

Oral antibody to interleukin-10(anti-IL-10)enhances the intestinal immune defense against Eimeria.The sulfur amino acids methionine and cysteine(M+C)play essential roles in inducing and maintaining protective immune responses during intestinal infections.Hence,increased dietary M+C may support the anti-IL-10-induced intestinal immunity to Eimeria.Broilers(n=640)were arranged in a 2×2×2 factorial design with 2 levels of each of the 3 main factors:dietary standardized ileal digestible(SID)M+C levels(0.6％or 0.8％),dietary anti-IL-10 supplementation(with or without),and coccidiosis chal-lenge(control or challenge).Briefly,the broilers were supplied with either 0.6％or 0.8％SID M+C,each with or without anti-IL-10(300 μg/kg),from d 10 to 21.On d 14,broilers from each diet were gavaged with either PBS or Eimeria.The resulting Eimeria infection induced fecal oocyst shedding and intestinal lesions.Broilers fed 0.8％SID M+C(main effects,P≤0.05)had decreased feed-to-gain ratio,increased duodenum and cecum luminal anti-Eimeria IgA titers,and decreased fecal oocyst counts,when compared to 0.6％SID M+C.The supplementation of anti-IL-10(main effects,P≤0.05)increased cecum luminal total IgA concentration and decreased cecum lesions.Interactions(P≤0.05)were detected for growth performance and cecum luminal IFN-γ.Briefly,the highest body weight gain and feed intake were reached in PBS-gavaged broilers fed 0.8％SID M+C with no anti-IL-10 and in Eimeria-challenged broilers fed 0.8％SID M+C with anti-IL-10.In Eimeria-infected broilers,anti-IL-10 increased intestinal luminal IFN-γ and body weight gain only at 0.8％SID M+C.Collectively,anti-IL-10 increased intestinal luminal IFN-γ levels,decreased cecum lesions and restored growth only when fed with adequate amounts of sulfur amino acids.Our findings underscore the importance of providing sufficient essential nutrients to support the anti-IL-10 induced immunity against coccidiosis.

Experiments were conducted to study the effect subtherapeutic levels of dietary antibiotic supplementation on the growth, intestinal characteristics and bacterial populations of chicks reared on "dirty" versus "clean" water. the lack of growth response to antibiotics in a Results showed that growth response to penicillin was independent of water treatment and the magnitude of growth response ranged from 0.9 to 18.3% levels. Bursal weights were significantly increased (P<0.05) and intestinal weights significantly decreased (P<0.01) in. chicks fed penicillin. The effect of antibiotics on The lack of growth response to antibiotics in intestinal thickness and length were inconsistent while bacterial counts on Sulfite - the only common denominator to all Polymyxin-Sulfadiazine (SPS) and K.F. antibiotics is their ability to inhibit the growth streptococcus agar were unaffected by penicillin and water treatments.

Targeting fibroblast growth factor 23 (FGF-23) signaling pathway is of interest in controlling body phosphate metabolism. This study investigated the effect of anti-fibroblast growth factor receptor 1 (FGFR1, major FGF-23 receptor in the kidney) antibodies on phosphate metabolism. White Leghorn laying hens (65-wk-old) were vaccinated with either a FGFR1 peptide vaccine (five 8-amino-acid peptides were selected, CrZ-1:LPEDPRWE, CrZ-2:LDKDKPNR, CrZ-3:RRPPGMEY, CrZ-4:GSPYPGVP, and CrZ-5:RMDKPSNC) or adjuvant control. At peak antibody titer, hens were artificially inseminated. Chicks from control-vaccinated hens were fed either a non-phytate phosphorus (nPP) sufficient (nPP = 0.45%, positive control) or deficient (nPP = 0.20%, negative control) diet, while chicks from each of the FGFR1 peptide vaccinated hens were fed with the above nPP-deficient diet, for 14 D. When compared to control hens, plasma phosphate in CrZ-1, CrZ-2, CrZ-3, CrZ-4, and CrZ-5 vaccinated hens were decreased by 33, 30, 24, 20, and 26%, respectively (P < 0.05); egg weight in CrZ-2 and CrZ-5 vaccinated hens were increased by 6 and 7%, respectively (P < 0.05); egg production in CrZ-3, CrZ-4, and CrZ-5 vaccinated hens tended to decrease (P = 0.085; decreased by 14, 15, and 13%, respectively). When compared to positive control, chicks from all other groups had decreased body weight gain (BWG) and feed intake (FI) during 1 to 14 D, and had decreased plasma phosphate, tibiotarsus ash, and 24-h phosphorus excretion on day 14. When compared to negative control, BWG of CrZ-1, CrZ-2, CrZ-3, and CrZ-4 antibody chicks were decreased by 23, 28, 26, and 20%, respectively (P < 0.05); FI of CrZ-1, CrZ-2, and CrZ-3 antibody chicks were decreased by 15, 15, and 18%, respectively (P < 0.05); plasma phosphate of CrZ-5 antibody chicks were decreased by 26% (P < 0.05); plasma FGF-23 levels of CrZ-4 antibody chicks were increased by 18% (P < 0.05); tibiotarsus ash content of CrZ-2, CrZ-3, and CrZ-4 antibody chicks were decreased by 20, 20, and 21%, respectively (P < 0.05). In conclusion, anti-FGFR1 peptide antibodies decreased egg production of hens and growth performance of their progeny chicks probably by activating FGF-23 signaling and stimulating FGF-23 production.

Eimeria species are intestinal protozoan parasites that cause lack of production, malabsorption and mortality in floor raised chickens. Administering an oral antibody to interleukin 10 (aIL-10) reduces the symptoms of coccidiosis in broilers, indicating interleukin 10 (IL-10) is key to Eimeria pathology. IL-10 is an anti-inflammatory cytokine and acts as a stand down signal to reduce inflammation and host pathology during disease. Related protozoan parasites exploit IL-10 to reduce pathogen-damaging host inflammatory responses. We hypothesize that IL-10 is increased during Eimeria infection through an unknown host-pathogen interaction, and by feeding aIL-10 to neutralize excess IL-10 the bird is allowed to mount an effective immune response to Eimeria. To determine the effects of aIL-10 during the intestinal immune response, intestinal pathology and the relationship between IL-10, interferon gamma (IFNγ) and Eimeria infection were evaluated in this study. In both experiments, broilers were administered either a 10x dose of Advent® Eimeria vaccine or saline. Duodenum, jejunum and cecum samples were collected, processed, stained and examined under a microscope. Evaluation of intestinal histomorphology during aIL-10 administration showed minimal differences in birds fed aIL-10 during infection compared to animals fed a control antibody during Eimeria infection. To further evaluate aIL-10's positive effect during infection, immunofluorescent histochemistry was performed on chicken intestines days 3-7 post Eimeria infection for IL-10 and IFNγ presence in intestinal mucosa in control and infected birds, in regions with and without visible Eimeria burden. IL-10 and IFNγ had significant changes between days 4.5-7 post-infection in birds fed aIL-10 compared to animals fed a control antibody. Overall we found that the duodenum had increased IL-10 presence and increased IFNγ presence, and the jejunum and cecum had decreased IL-10 presence and decreased IFNγ presence. These differences in spatial regulation of IL-10 and IFNγ may indicate Eimeria species induce slightly different cytokine responses.

Coccidiosis is a major gastrointestinal disease caused by several Eimeria species in floor raised chickens. Feeding an antibody to interleukin 10 (aIL-10) ameliorates the negative symptoms of coccidiosis in broilers, i.e., lack of weight gain, decreased feed conversion, and mortality. IL-10 signals by forming a ligand-receptor complex with IL-10 Receptor 1 (IL-10 R1) and IL-10 Receptor 2 (IL-10 R2). In this study, we hypothesize oral antibodies to the IL-10 receptors will neutralize the IL-10 signaling pathway equal to or better than aIL-10 to act as an oral anti-coccidiosis immunotherapy. A total of 5 sequential feed trials, set up as a 4 (diet antibody) × 2 (Eimeria challenge) factorial design, tested oral egg yolk antibodies to a total of 6 IL-10 R1 epitopes and 3 IL-10 R2 epitopes compared to a control antibody diet. A total of 10 pens of 5 chicks/pen/diet antibody/Eimeria challenge were housed for 21 d. On day 3 of age, chicks were either infected or not infected with a 10× dose of an Eimeria vaccine containing Eimeria acervulina, Eimeria tenella, and Eimeria maxima. Pen feed consumption and mean body weights were assessed weekly (d1, d7, d14, and d21); fecal oocyst shedding was assessed on day 10. Data were analyzed using a 2-way ANOVA. No significant interaction on chick weight was observed in chicks fed IL-10 R1 antibodies compared to chicks fed the control antibody was observed. In studies evaluating aIL-10 R2 oral antibodies, infected chicks fed aIL-10 R2: epitope 1 overcame the negative effects of Eimeria infection and had similar 21-d body weight to uninfected chicks (P4 = 0.07). We hypothesized that feeding oral antibodies to the IL-10 receptors would result in equivalent anti-coccidial benefits to aIL-10. However, none of the 6 antibodies to IL-10 R1 epitopes yielded any benefits during Eimeria infection compared to controls. A total of 2 oral antibodies to IL-10 R2 showed promising results equivalent to the aIL-10 immunotherapeutic. Immunofluorescence staining shows that the IL-10R2 significantly increases in abundance in response to Eimeria infection, whereas IL-10R1 does not.

Subunit vaccines can have excellent safety profiles, but their ability to give rise to robust immune responses is often compromised. For glycan-based vaccines, insufficient understanding of B and T cell epitope combinations that yield optimal immune activation hinders optimization. To determine which antigen features promote desired IgG responses, we synthesized epitope-functionalized polymers using ring-opening metathesis polymerization (ROMP) and assessed the effect of B and T cell epitope loading. The most robust responses were induced by polymers with a high valency of B and T cell epitopes. Additionally, IgG responses were greater for polymers with T cell epitopes that are readily liberated upon endosomal processing. Combining these criteria, we used ROMP to generate a nontoxic, polymeric antigen that elicited stronger antibody responses than a comparable protein conjugate. These findings highlight principles for designing synthetic antigens that elicit strong IgG responses against inherently weak immune targets such as glycans.

Yellow and orange egg yolks are good sources of xanthophyll carotenoids, consumption of which is associated with health benefits, such as cancer prevention, eye health, and bone health. Industrial feed fortificants used to improve egg yolk color and carotenoid concentration typically are derived from marigold flowers. Green leafy vegetables are also concentrated sources of the xanthophylls lutein and zeaxanthin (L+Z), but they have not been rigorously evaluated in laying hen feeds as a yolk colorant. The addition of food manufacturing byproducts, including carrot leaves, to animal feed is a promoted method of improving animal nutrition. The ability of dehydrated carrot leaves to improve egg yolk color and L+Z concentration was evaluated by feeding laying hens (n = 40) white maize-based feeds fortified with 2 different dehydrated carrot leaves, marigold as a positive control, or no fortificant as a negative control for 28 D. After a 7-D washout period, the hens were separated into 4 groups, and eggs were collected every other day. Yolks were analyzed by using a portable colorimeter to define the color space and by ultra-performance liquid chromatography to determine the carotenoid profile. Carotenoid concentration rapidly declined from day 0 to 8, confirming adequate washout conditions. The white maize negative control (WM) day 28 lutein concentration (3.59 ± 0.51 nmol/g) was significantly less than orange-carrot leaf-treated (OCL) (5.34 ± 0.36 nmol/g) and red-carrot leaf-treated hens (RCL) (5.92 ± 1.00 nmol/g) in addition to the marigold-treated hens (MG). However, MG was significantly higher than both leaf-treated groups. From day 8 (3.93 ± 0.74 nmol/g) to 28 (9.32 ± 1.66 nmol/g), MG had the largest increase in lutein and was the only treatment to surpass day 0 initial concentrations (8.50 ± 1.64 nmol/g). A similar trend was observed for zeaxanthin and was reflected in the color space.

Research has shown that methionine+ cysteine (M+C) requirements may be higher when chickens are infected with Eimeria app. In a 4 × 2 factorial design, broilers (11 to 21 D) were fed one of 4 corn–soybean meal-based diets containing either 0.6, 0.8, 0.9, or 1.0% standardized ileal digestible (SID) M+C; on day 14, broilers from each diet were gavaged with either phosphate-buffered saline (PBS) or a commercial coccidiosis vaccine (at 100 × vaccine dose) which provide a mixture of live Eimeria acervulina, Eimeria maxima, and Eimeria tenella oocysts. Growth performance was recorded from day 11 to 21. Plasma and intestinal luminal samples were collected on days 14 and 21. Intestine lesion scores and fecal oocyst counts were conducted on day 21. Regardless of dietary SID M+C levels, compared to PBS gavaged broilers, the Eimeria-challenged broilers had (1) decreased (P < 0.05) body weight gain (BWG), feed intake (FI), and gain-to-feed ratio (G:F); (2) increased (P < 0.05) intestinal lesion scores and fecal oocyst counts; (3) increased (P < 0.05) plasma anti-Eimeria IgG, and intestinal luminal total IgA and anti-Eimeria IgA concentrations; and (4) increased (P < 0.05) levels of duodenum luminal gamma interferon (IFN-γ) and interleukin-10 (IL-10), as well as jejunum and cecum luminal IFN-γ concentrations. Regardless of Eimeria challenge, when compared to 0.6% SID M+C, broilers fed ≥0.8% SID M+C had (1) increased (P < 0.05) BWG, FI, and G:F and (2) increased (P < 0.05) levels of jejunum luminal total IgA. After Eimeria challenge, broilers fed 0.8% SID M+C had increased (P < 0.05) levels of jejunum luminal anti-Eimeria IgA compared to broilers fed diets containing 0.6 and 1.0% SID M+C. Collectively, in 11- to 21-D broilers, the growth suppression caused by Eimeria infection could not be mitigated by further increasing dietary M+C alone ≥0.8%. Further research should investigate interactions between dietary M+C and other nutrients for support of immune function and growth in pathogen-challenged broilers.

Dietary factors such as adenine have been linked to phosphate-calcium metabolism disturbance and adverse productive outcomes. Anti-fibroblast growth factor 23 (FGF-23) antibody has been proposed to ameliorate adenine-induced abnormal FGF23/phosphate metabolism. This experiment was conducted to investigate the application of anti-FGF-23 antibody in adenine-gavaged laying hens. Single Comb White Leghorn laying hens with (n = 10) or without (control group, n = 10) systemic anti-FGF-23 antibody were orally gavaged with adenine (600 mg/hen/D) for 21 consecutive days. Adenine gavage increased (P ≤ 0.01) plasma phosphate and calcium levels and tended to increase (0.05 < P ≤ 0.1) plasma 1,25-dihydroxy-cholecalciferol [1,25(OH)2D3] level of hens without FGF-23 antibody. In hen with anti-FGF-23 antibody, adenine gavage increased (P ≤ 0.01) body weight and plasma calcium level and decreased (P ≤ 0.05) plasma FGF-23 level. Feed intake of hens in both treatments was suddenly decreased (control hens decreased from 111 to 55 g, P ≤ 0.01; anti-FGF-23 hens decreased from 96 to 46 g, P ≤ 0.01) 10 D after adenine gavage. Anti-FGF-23 antibody tended to increase (0.05 < P ≤ 0.1) plasma phosphorus level of hens before adenine gavage, interestingly, and decreased (P ≤ 0.01) plasma FGF-23 level and kidney index (% of body weight) of hens after adenine gavage. In conclusion, anti-FGF-23 antibody might be used (before or in the early stage) to delay the development of adenine-induced abnormal FGF23/phosphate metabolism. This is the first study to investigate the FGF-23 status in chickens suffering from dietary factors which may cause abnormal renal phosphate resorption.

The primary objective of this randomized controlled trial was to determine whether anti-IL-10 egg yolk antibodies fed upon arrival to a calf ranch would lower the prevalence of Cryptosporidium parvum shedding in naturally challenged preweaned dairy calves. The secondary objectives included measuring the effect of anti-IL-10 antibodies on calf health, performance, and shedding of less common diarrheal pathogens. A total of 133 calves, enrolled at 24 to 72 h of age, received a daily dose of 0.96 g of egg yolk powder with anti-IL-10 antibodies (MAB, n = 71) or without anti-IL-10 antibodies (MEP, n = 62) split between 2 feedings for the first 11 d on feed at a calf ranch. Daily health evaluations were completed for 15 d after arrival and on d 56. Digital weights were collected at enrollment and d 56, and hipometer weights were collected at enrollment and d 7 and 56. Packed cell volume and serum total protein concentration were measured at enrollment and on d 7 and 14. Fecal pH was measured at enrollment and on d 5 and 14, and fecal pathogen (C. parvum, coronavirus, rotavirus, and Salmonella spp.) shedding was assessed at d 5 and 14. Continuous outcomes were compared between groups using a Student's t-test or Wilcoxon rank sum test. Fecal pathogen shedding at d 14, respiratory disease at d 56, and antibiotic usage were compared using relative risk (RR) and chi-squared test. Fecal pH (median and interquartile range) on d 14 was 6.65 (6.39–6.99) and 6.52 (5.97–6.81) for MAB and MEP, respectively. On d 56, the risk of respiratory disease was lower for MAB compared with MEP (RR = 0.40; confidence interval = 0.16–0.99). The risk for antibiotic treatment was lower for MAB- compared with MEP-treated calves (RR = 0.38; confidence interval = 0.17–0.88). The risk of shedding rotavirus was higher in MAB (RR = 1.38; confidence interval = 1.10–1.81) calves. After multivariable analyses, hipometer weights (least squares means ± standard error) were 1.7 ± 0.8 kg greater on d 56 in MAB compared with MEP; however, ADG was 0.04 ± 0.02 kg/d lower in MAB calves. Total health score, diarrhea days, average respiratory score, packed cell volume, and serum total protein were not affected by feeding anti-IL-10 egg antibodies. In summary, feeding anti-IL-10 antibodies was associated with increased fecal pH, reduced risk of respiratory disease later in the preweaning period, and decreased antibiotic usage despite higher rotavirus infection. These findings might be associated with improved mucosal immunity, enhanced host defenses, or reduced susceptibility and warrant further investigation.

The goal of this study was to investigate the anti-cancer effects of Trans10,cis12 conjugated linoleic acid (t10,c12 CLA). MTT assays and QCM™ chemotaxis 96-wells were used to test the effect of t10,c12 CLA on the proliferation and migration and invasion of cancer cells. qPCR and Western Blotting were used to determine the expression of specific factors. RNA sequencing was conducted using the Illumina platform and apoptosis was measured using a flow cytometry assay. t10,c12 CLA (IC50, 7 μM) inhibited proliferation of ovarian cancer cell lines SKOV-3 and A2780. c9,t11 CLA did not attenuate the proliferation of these cells. Transcription of 165 genes was significantly repressed and 28 genes were elevated. Genes related to ER stress, ATF4, CHOP, and GADD34 were overexpressed whereas EDEM2 and Hsp90, genes required for proteasomal degradation of misfolded proteins, were downregulated upon treatment. While apoptosis was not detected, t10,c12 CLA treatment led to 9-fold increase in autophagolysosomes and higher levels of LC3-II. G1 cell cycle arrest in treated cells was correlated with phosphorylation of GSK3β and loss of β-catenin. microRNA miR184 and miR215 were upregulated. miR184 likely contributed to G1 arrest by downregulating E2F1. miR215 upregulation was correlated with increased expression of p27/Kip-1. t10,c12 CLA-mediated inhibition of invasion and migration correlated with decreased expression of PTP1b and decreased Src activation by inhibiting phosphorylation at Tyr416. Due to its ability to inhibit proliferation and migration, t10,c12 CLA should be considered for treatment of ovarian cancer.

Cryptosporidium parvum infection is a nearly ubiquitous infection in dairy herds for which there are no approved treatments or vaccines available in the US. Therefore, oral administration of immunoglobulins to control enteric disease in calves is of significant interest. Antibodies survive gastrointestinal transit, are accessible in the intestinal lumen, and remain locally active without the risk of violative residues. Our research team previously demonstrated that calves fed anti-11-10 IgY in milk had higher fecal pH, reduced respiratory disease, and increased frame measurements. Therefore, the objective of the current study was to carry out a dose-response trial evaluating the effects of feeding 0.5x, 2.0x, and lx the dose from the original study on respiratory and gastrointestinal health, C. parvum oocyst shedding, and calf growth.

Mixed-isomer conjugated linoleic acid (CLA) and the individual isomers, trans-10, cis-12 (CLAt10c12) and cis-9, trans-11 (CLAc9t11), decrease severity of collagen-induced arthritis (CA) when consumed after disease onset. Few studies have been conducted exploring the role of CLA in the prevention of autoimmune diseases. These studies suggest that isomer-specific effects may be occurring; however, a direct comparison of CLAt10c12 and CLAc9t11 has yet to be conducted. A study to compare the ability of CLAt10c12 and CLAc9t11 to prevent CA and assess their effects on early inflammation was performed. DBA/1 mice were fed a semipurified diet containing 6% corn oil (CO), 5.5% CO and 0.5% CLAt10c12, or 5.5% CO and 0.5% CLAc9t11 (n = 27 per diet) starting three weeks before CA primary immunization. Effects on disease incidence and severity, anticollagen antibodies, plasma and paw cytokines, and hepatic fatty acids were measured. Arthritis incidence was reduced by a minimum of 34% in mice fed either CLA isomer compared to those fed CO diet (p = 0.06). In mice that did develop arthritis (n = 9-12 mice per treatment), CLAt10c12 reduced arthritic severity to a greater extent than CLAc9t11 and CO (p = 0.03). CLA isomer treatment attenuated the increased hepatic arachidonic acid (ARA; 20:4n-6) observed with arthritis at one-week postonset (p = 0.03), while no differences in anticollagen antibodies or cytokines were observed between dietary treatments. These results suggest that CLA isomers may be effective at preventing specific immune-mediated inflammatory diseases, in part, through modulation of the ARA cascade.

Strategies that would increase eggshell quality could be of considerable value to egg producers. This research demonstrated the effective use of fibroblast growth factor 23 (FGF-23) peptide vaccines to increase eggshell quality of Single Comb White Leghorn laying hens (from 69 to 72 wk of age). Hens, fed a standard diet (containing 900 IU/kg vitamin D3), were intramuscularly injected (and boosted) with either a control vaccine (n = 14 hens) or one of 2 FGF-23 peptide vaccines (peptides NP1, GMNPPPYS; and NP7, YTSTERNSFH; n = 15 hens for each peptide). During peak antibody titer, eggs were collected for shell and internal quality analysis, hens were artificially inseminated, and the hatchability of fertilized eggs was determined. Laying hens vaccinated with either FGF-23 peptide NP1 or NP7 had increased (P < 0.05) plasma phosphate level (mmol/L; NP1 = 1.74, NP7 = 1.76, control = 1.47), egg specific gravity (NP1 = 1.083, NP7 = 1.083, control = 1.079), and eggshell strength (g of force; NP1 = 4002, NP7 = 4157, control = 3102) when compared to control vaccinated hens. FGF-23 peptide NP1 vaccinated hens also had increased eggshell thickness (mm, P < 0.001), shell weight (g, P = 0.032), and shell index (% of whole egg, P = 0.023) when compared to control vaccinated hens. FGF-23 peptide NP7 vaccinated hens tended to have decreased eggshell weight (P = 0.064) when compared to control vaccinated hens. Hatchability of fertilized eggs was not affected in incubations 1 and 3, but tended to be decreased (P = 0.097) by FGF-23 peptide NP1 vaccination in incubation 2. In conclusion, vaccines to FGF-23 peptides increased eggshell quality of laying hens with minimal adverse effects on egg internal quality. The effect of FGF-23 peptide vaccination on hatchability remains to be clarified.