The paradoxical effects of progesterone on the eggshell quality of laying hens

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 (P₄-2 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 (P₄-5 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 P₄-2 h treated hens whereas Val and Lys were significantly lower (P < 0.05) in P₄-5 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. P₄ injected affected the content of amino acids in eggshell membranes, especially lysine which contributed to eggshell quality. In addition, P₄ 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.     

Validated quantitative trait loci for eggshell quality in experimental and commercial laying hens

Compromised eggshell quality causes considerable economic losses for the egg industry. Breeding for improved eggshell quality has been very challenging. Eggshell quality is a trait that would greatly benefit from marker‐assisted selection, which would allow the selection of sires for their direct contribution to the trait and would also allow implementation of measurements integrating a number of shell parameters that are difficult to measure. In this study, we selected the most promising autosomal quantitative trait loci (QTL) affecting eggshell quality on chromosomes 2, 3, 6 and 14 from earlier experiments and we extended the F₂ population to include 1599 F₂ females. The study was repeated on two commercial populations: Lohmann Tierzucht Rhode Island Red line (n = 692 females) and a Hy‐Line White Plymouth Rock line (n = 290 progeny tested males). We analyzed the selected autosomal QTL regions on the three populations with SNP markers at 4–13 SNPs/Mb density. QTL for eggshell quality were replicated on all studied regions in the F₂ population. New QTL were detected for eggshell color on chromosomes 3 and 6. Marker associations with eggshell quality traits were validated in the tested commercial lines on chromosomes 2, 3 and 6, thus paving the way for marker‐assisted selection for improved eggshell quality.     

Hyperphosphataemia and bone resorption in histamine injected laying hens

The effects of histamine on bone resorption during eggshell formation, were studied. The study included the effect of histamine on the clearance of 45Ca and 32P from plasma, and on the release from the femur or uptake by the egg shell of 45Ca and 32P injected at the time of the first experiment or 10 hr prior to the experiment (experiment 2). Histamine led to a decrease in the disappearance rates of 45Ca and 32P, without any modification of the femur and egg shell levels. However, the increase in the amount of plasma 32P and the decrease in the 32P specific activity (SA) in bone, both elicited by histamine, argues in favour of the skeletal origin of hyperphosphataemia induced by histamine during egg shell formation.     

Early rearing enrichments influenced nest use and egg quality in free-range laying hens

In Australia, free-range egg production pullets are typically reared indoors, but adult layers get outdoor access. This new environment may be challenging to adapt to, which could impair egg production and/or egg quality. Adaptation might be enhanced through rearing enrichments. We reared 1386 Hy-Line Brown® chicks indoors with three treatments across 16 weeks: (1) a control group with standard litter housing conditions, (2) a novelty group providing novel objects that changed weekly, and (3) a structural enrichment group with custom-designed structures to partially impair visibility across the pen and allow for vertical movement. Pullets were transferred to a free-range system at 16 weeks of age with daily outdoor access provided from 25 until 64 weeks. Daily egg production at different laying locations (large nests, small nests and floor), weekly egg weights and egg abnormalities were recorded from 18 to 64 weeks old. External and internal egg quality parameters of egg weight, shell reflectivity, albumen height, haugh unit, yolk colour score, shell weight and shell thickness were measured at 44, 52, 60 and 64 weeks. There was a significant interaction between rearing treatment and nest box use on hen-day production from weeks 18 to 25 (P < 0.0001) with the novelty hens laying the most eggs and the control hens the fewest eggs in the nest box. Similarly, from 26 to 64 weeks, the novelty hens laid more eggs in the large nest boxes and fewer eggs on the floor than both the structural and control hens (P < 0.0001). Egg weight and abnormalities increased with age (P < 0.0001), but rearing treatment had no effect on either measure (both P ≥ 0.19). Rearing treatment affected shell reflectivity and yolk colour with the control hens showing paler colours across time relative to the changes observed in the eggs from enriched hens. The novelty hens may have established nest box laying patterns as they were more accustomed to exploring new environments. The differences in egg quality could be related to stress adaptability or ranging behaviour. This study shows that enriching environments during rearing can have some impacts on production parameters in free-range hens.     

Dietary supplementation with sodium bicarbonate or sodium sulfate affects eggshell quality by altering ultrastructure and components in laying hens

Dietary sodium (Na) levels were related to the content of the eggshell matrix. We therefore speculated that dietary Na supplementation as sodium bicarbonate (NaHCO₃) or sodium sulfate (Na₂SO₄) may improve eggshell quality. Additionally, dietary NaHCO₃ or Na₂SO₄ supplementation may further affect eggshell quality in different ways due to differences in anions. This study investigated and compared the effects of dietary Na supplementation in either NaHCO₃ or Na₂SO₄ form on laying performance, eggshell quality, ultrastructure and components in laying hens. A total of 576 29-week-old Hy-Line Brown laying hens were randomly allocated to 8 dietary treatments that were fed a Na-deficient basal diet (0.07% Na, 0.15% Cl) supplemented with Na₂SO₄ or NaHCO₃ at 0.08, 0.18, 0.23 or 0.33% Na for 12 weeks. No differences were observed in laying production performance with dietary Na supplementation. Dietary Na supplementation resulted in quadratic increases of eggshell breaking strength in both Na₂SO₄ and NaHCO₃ added groups (P < 0.05), and Na₂SO₄-fed groups had a quadratic increase in the eggshell ratio at week 12 (P < 0.05). Compared with supplementing 0.08% Na, dietary supplementation of 0.23% Na increased the effective thickness (P < 0.05) in both Na₂SO₄ and NaHCO₃ added groups, but decreased the thickness and knob width of the mammillary layer (P < 0.05). A linear increase on the calcium content of the shell was only observed with Na supplementation from NaHCO₃ (P < 0.05). No differences were observed in Na contents of the shell with dietary Na supplemented by both sources. Dietary Na addition had a quadratic increase on uronic acid contents of shell membrane in NaHCO₃-fed groups (P < 0.05). Moreover, the sulfated glycosaminoglycan (GAG) contents of shell membranes increased linearly with dietary Na supplementation (P < 0.05). Dietary supplementation of 0.23% Na from Na₂SO₄ increased the sulfated GAG contents of calcified eggshell (P < 0.05). Additionally, compared with NaHCO₃-fed groups, Na₂SO₄-fed groups had higher eggshell breaking strength, thickness, eggshell weight ratio, effective thickness and the sulfated GAG contents of calcified eggshell at week 12. Overall, dietary supplementation of NaHCO₃ or Na₂SO₄ could increase eggshell breaking strength, which may be related to increased sulfated GAG contents in eggshell membranes and improved ultrastructure. Higher eggshell breaking strength, thickness and eggshell ratio could be obtained when the diet was supplemented with 0.23% Na from Na₂SO₄.     

Relationships among age, eggshell thickness and vitamin D metabolism and its expression in the laying hen.

Hens forming uncalcified shells synthesized less 1,25-hydroxycholecalciferol (1,25(OH)2D3) and less duodenal and eggshell gland (ESG) calbindin than normal laying hens. Hens forming thin shells had lower intestinal and ESG calbindin and its mRNA. Reducing ESG calcium (Ca2+) transport by the carbonic anhydrase inhibitor acetazolamide, but not by dietary Ca2+ restriction, reduced ESG calbindin and its mRNA. Two sub-populations of hens characterized by shell thickness (ST) maintained this characteristic throughout the whole production period. The differences between the two sub-populations increased with age. In old laying hens, the two sub-populations responded differently to dietary Ca2+ restriction and to exogenous 1,25(OH)2D3. Those forming a thin shell responded to 1,25(OH)2D3 by a significant improvement in ST. The results suggest that: (a) the mechanism responsible for Ca2+ transport to the egg shell consists of a vitamin D-dependent absorption of Ca2+ and a multi-factor-dependent transfer of Ca2+ to the shell; (b) both steps are, most likely, calbindin-mediated; however, the induction of calbindin gene expression in the ESG is predominantly calcium-dependent; and (c) the apparent defect in vitamin D metabolism or its expression in old hens is typical of, or even exclusive, to thin-shell-forming hens.     

Low levels of organic compound trace elements improve the eggshell quality,antioxidant capacity,immune function,and mineral deposition of aged laying hens

In the egg production industry, trace elements are required as additional dietary supplements to play vital roles in performance and egg quality. Compared to inorganic microelements (ITs), appropriate dose of organic trace microelements (OTs) are environmentally friendly and sufficient to satisfy the needs of hens. In order to evaluate the extent to which low-dose OTs replace whole ITs, the effects of organic copper, zinc, manganese, and iron compound on the performance, eggshell quality, antioxidant capacity, immune function, and mineral deposition of old laying hens were investigated. A total of 1 080 57-week-old Jing Hong laying hens were assigned to five groups with six replicates of 36 layers each for an 8-week experimental period. The birds were fed either a basal diet (control treatment (CT)) or the basal diet supplemented with commercial levels of inorganic trace elements (IT 100%) or the equivalent organic trace elements at 20%, 30%, and 50% of the inorganic elements (OT 20%, OT 30%, and OT 50%, respectively). Results showed that compared with those in the CT treatment, feeding hens with inorganic or organic microelement diet had significant effects on the eggshell quality, antioxidant capacity, immune function, and mineral deposition of old laying hens (P < 0.05). The eggshell strength and ratio between OT 30%, OT 50%, and IT 100% were similar at weeks 4 and 8, and the eggshell thickness of these groups was also similar at weeks 6 and 8. At week 8, the eggshell colour in OT 50% was darker than that in IT 100%. The mineral content in the eggshells of OT 50% and IT 100% significantly increased (P < 0.001), with no significant difference in effective thickness, mammillary thickness, and mammillary knob width between groups. There were no differences in the malondialdehyde content, total antioxidant capacity, and total superoxide dismutase activity in serum between OT 30%, OT 50%, and IT100%. While the catalase activities, the interleukin-1β, interleukin-10, immunoglobulin G, and immunoglobulin M concentrations in serum were not significantly different between OT 50% and IT 100%. The mineral contents in the faeces of the organic groups were considerably reduced compared with those in IT 100% (P < 0.001). In conclusion, dietary supplementation with 30–50% organic compound microelements has the potential to replace 100% inorganic microelements in the hen industry for improving eggshell quality, mineral deposition in the eggshell, antioxidant capacity, and immune function, and reducing emissions to the environment without negative effects on laying performance.     

Influence of physical activity on tibial bone material properties in laying hens

Laying hens develop a type of osteoporosis that arises from a loss of structural bone, resulting in high incidence of fractures. In this study, a comparison of bone material properties was made for lines of hens created by divergent selection to have high and low bone strength and housed in either individual cages, with restricted mobility, or in an aviary system, with opportunity for increased mobility. Improvement of bone biomechanics in the high line hens and in aviary housing was mainly due to increased bone mass, thicker cortical bone and more medullary bone. However, bone material properties such as cortical and medullary bone mineral composition and crystallinity as well as collagen maturity did not differ between lines. However, bone material properties of birds from the different type of housing were markedly different. The cortical bone in aviary birds had a lower degree of mineralization and bone mineral was less mature and less organized than in caged birds. These differences can be explained by increased bone turnover rates due to the higher physical activity of aviary birds that stimulates bone formation and bone remodeling. Multivariate statistical analyses shows that both cortical and medullary bone contribute to breaking strengthThe cortical thickness was the single most important contributor while its degree of mineralization and porosity had a smaller contribution. Bone properties had poorer correlations with mechanical properties in cage birds than in aviary birds presumably due to the greater number of structural defects of cortical bone in cage birds.     

Investigating social discrimination of group members by laying hens

Social relationships in domestic fowl are commonly assumed to rely on social recognition and its pre-requisite, discrimination of group-mates. If this is true, then the unnatural physical and social environments in which commercial laying hens are typically housed, when compared with those in which their progenitor species evolved, may compromise social function with consequent implications for welfare. Our aims were to determine whether adult hens can discriminate between unique pairs of familiar conspecifics, and to establish the most appropriate method for assessing this social discrimination. We investigated group-mate discrimination using two learning tasks in which there was bi-directional exchange of visual, auditory and olfactory information. Learning occurred in a Y-maze task (p < 0.003; n = 7/8) but not in an operant key-pecking task (p = 0.001; n = 1/10). A further experiment with the operant-trained hens examined whether failure was specific to the group-mate social discrimination or to the response task. Learning also failed to occur in this familiar/unfamiliar social discrimination task (p = 0.001; n = 1/10). Our findings demonstrate unequivocally that adult laying hens kept in small groups, under environmental conditions more consistent with those in which sensory capacities evolved, can discriminate group members: however, appropriate methods to demonstrate discrimination are crucial.     

Short-term feather maintenance as a function of dust-bathing in laying hens

Laying hens, deprived of dust for 33 days, showed an average increase in amount of lipids on back feathers from 10.3 to 14.5 mg lipids per g feathers at the end of the dust-deprivation period. After the hens could dust-bathe again, the original level was restored within 2 days. Also the downy parts of these feathers appeared to be fluffier. Dust-bathing presumably regulates the amount of feather lipids and maintains down structure in good condition. These conclusions are discussed in relation to earlier findings concerning feather lipid quantity and feather structure.     

Zearalenone, deoxynivalenol and fumonisins in mixed-feed for laying hens

Fusarium toxins are secondary metabolites produced byfungi of these genera in many commodities under certain conditions. A study was carried out to investigate the co-occurrence of zearalenone (ZEN), deoxynivalenol (DON) and fumonisins (FB₁ and FB₂) in 52 samples of mixed-feed for poultry contaminated withFusarium verticillioides. The zearalenone and deoxynivalenol were checked using immunoaffinity column and the extraction of fumonisin was performed by strong anion exchange (SAX) solid phase column. Detection and quantification were determined by high performance liquid chromatography (HPLC). The limit of detection was 5 μg/kg for ZEN, 100 μg/kg for DON and 50 and 100 μg/kg for FB₁ and FB₂ respectively.Fusarium toxins were detected in 20 samples. Sixteen samples were positive for ZEN (30.7%) presenting levels that ranged from 7.4 μg/kg to 61.4 μg/kg (mean=27.0 μg/kg). 13.5% of the samples presented contaminations of DON, with levels ranging from 100.0 μg/kg to 253 μg/kg (mean=l18.07 μg/kg). FB₁ was detected in 19.2% of samples, with levels ranging from 50.0 μg/kg to 110.0 μg/kg (mean=73.6 μg/kg). FB2 was not detected in any sample. In positive samples simultaneously contamination with two or three mycotoxins were detected in 9 of them (17.3%).     

Effects of probiotic supplementation on performance traits,bone mineralization,cecal microbial composition,cytokines and corticosterone in laying hens

Recent researches have showed that probiotics promote bone health in humans and rodents. The objective of this study was to determine if probiotics have the similar effects in laying hens. Ninety-six 60-week-old White Leghorn hens were assigned to four-hen cages based on their BW. The cages were randomly assigned to 1 of 4 treatments: a layer diet mixed with a commercial probiotic product (containing Enterococcus faecium, Pediococcus acidilactici, Bifidobacterium animalis and Lactobacillus reuteri) at 0, 0.5, 1.0 or 2.0 g/kg feed (Control, 0.5×, 1.0× and 2.0×) for 7 weeks. Cecal Bifidobacterium spp. counts were higher in all probiotic groups (P<0.001) compared with the control group. The percentage of unmarketable eggs (cracked and shell-less eggs) was decreased in both 0.5× and 2.0× groups compared with the control group (P=0.02), mainly due to the reduction of shell-less eggs (P=0.05). The increases in tibial and femoral mineral density and femoral mineral content (P=0.04, 0.03 and 0.02, respectively), with a concomitant trend of increases in humerus mineral density and tibial mineral content (P=0.07 and 0.08, respectively), occurred in the 2.0× group. However, the bone remodeling indicators of circulating osteocalcin and c-terminal telopeptide of type I collagen were similar among all groups (P>0.05). In addition, the plasma concentrations of cytokines (interleukin-1β, interleukin-6, interleukin-10, interferon-γ and tumor necrosis factor-α) and corticosterone as well as the levels of heterophil to lymphocyte ratio were similar between the 2.0× group and the control group (P>0.05). In line with these findings, no differences of cecal tonsil mRNA expressions of interleukin-1β, interleukin-6 and lipopolysaccharide-induced tumor necrosis factor-α factor were detected between these two groups (P>0.05). These results suggest that immune cytokines and corticosterone may not involve in the probiotic-induced improvement of eggshell quality and bone mineralization in laying hens. In conclusion, the dietary probiotic supplementation altered cecal microbiota composition, resulting in reduced shell-less egg production and improved bone mineralization in laying hens; and the dietary dose of the probiotic up to 2.0× did not cause negative stress reactions in laying hens.