Influence of nest seclusion and nesting material on pre-laying behaviour of laying hens

Provision of nest sites is beneficial for the welfare of laying hens in intensive production systems. The design of these nest sites has a direct effect on pre-laying behaviour. The aim of this study was to investigate the effects of screening off the entrances of the nest boxes with non-transparent flaps and providing nesting or flooring material on pre-laying behaviour. Eighteen individual Hisex brown laying hens and 18 groups of five hens were tested for 2 days in a test cage containing three nest boxes each with a different nesting material (peat, artificial turf and plastic-coated wire mesh). On one of the days the nest boxes were equipped with plastic flaps at the entrances. Pre-laying behaviour was analysed using focal sampling. Both degree of seclusion and nesting material had an effect on pre-laying behaviour of hens. Nest boxes with flaps were visited less frequently but for a longer duration per visit than nest boxes without flaps. Hens spent more time sitting and scratching, and less time standing and moving when flaps were present. Hens also received fewer pecks (group test) when nest boxes were equipped with flaps. Major differences were found between pre-laying behaviour on coated wire mesh compared to peat and artificial turf with shorter duration of nest visits (group test), more standing (group test) and moving (individual and group test), and less sitting (individual and group test) and object pecking (group test) on coated wire mesh. Differences in pre-laying behaviour on peat versus artificial turf were observed for the behaviours moving (less on peat), sitting (more on peat in the individual test) and object pecking (more on peat in the group test).

It is concluded that hens show more settled pre-laying behaviour and more nest-building behaviour in the presence of flaps at the entrances of nest boxes. These results indicate that seclusion of nest boxes with non-transparent flaps is beneficial to laying hen welfare. Differences in pre-laying behaviour also suggest that plastic-coated wire mesh is less suitable as nesting material than peat and artificial turf.     

Influence of nest-floor slope on the nest choice of laying hens

Group nests in alternative housing systems for laying hens primarily fulfil the hen's needs for seclusion and protection. Commercial nests used in Switzerland are built according to the provisions of the Swiss Animal Welfare Legislation. However, nest types can differ in aspects, such as floor slope, that could have an impact on egg-laying behaviour. Floor slope has to be designed so that eggs roll away without breaking and so that hens feel comfortable laying their eggs. In commercial nests, the slope is usually between 12% and 18%. The aim of this study was to investigate the effect of floor slope on the hen's nest preference and laying behaviour. We predicted that hens would prefer nests with a lower sloped floor for evolutionary reasons and for reasons related to comfort.Eight pens, each with 17-18 white laying hens (LSL), were equipped with two roll-away nests (0.54 m²) having different floor slopes (12% and 18%). Eggs were collected each day (from approximately 20 weeks of age until 28 weeks of age); the number of eggs in each nest and on the floor of the pens was recorded. Behaviour inside the nest was filmed for two consecutive days during the main egg-laying time from the second hour to the fifth hour (4 h) after lights came on in week 27/28. The following data were recorded: number of hens in each nest, the nest visits/egg number ratio, the number of sitting events, the body alignment of hens sitting in the nest and the number and duration of nest visits. Data were analysed with a repeated-measures ANOVA. There was no difference between the numbers of eggs in the two nests, but more hens were counted in nests with a 12% slope (p = 0.027). The ratio between the number of nest visits and number of eggs did not differ significantly between the nests. However, we counted more sitting events in the nest with 12% slope (p = 0.007). The percentage of body alignment towards the back (p = 0.044) and towards the front (p = 0.028) of the nest differed between the nests. Furthermore, for nest visits lasting between 10 and 90 min, we found significant differences in the total number of nest visits (p = 0.039). For visits in this range of duration, we also found significant differences for nest visits with sitting (p = 0.025) and for the number of nest visits with egg laying (p = 0.049). All of these differences favoured the 12% nest.Both nests were generally accepted by the hens. However, because of the higher number of hens counted in the 12% nest and the higher amounts of nest visits and sitting events found in these nests, we recommend to use nests with a floor slope of 12% rather than 18%.     

Study of selected genes of Wnt signaling pathway in relation to the parameters in the bone tissue of the laying hens

The Wnt signaling pathway plays a critical role in almost all aspects of skeletal development and homeostasis. Many studies suggest the importance of this signaling pathway in connection with bone metabolism through many skeletal disorders caused by mutations in Wnt signaling genes. The knowledge gained through targeting this pathway is of great value for skeletal health and diseases, for example of increased bone mass in the case of osteoporosis. Our objective was to focus on the detection of single nucleotide polymorphisms and investigate the associations between possible polymorphisms in selected genes that are part of those signaling pathways and parameters of bones in hens of ISA Brown hybrids (bone breaking strength, length, width, and bone mass). Different regions of the GPR177, ESR1 and RUNX2 genes were studied, using PCR and sequencing, in a total of forty-eight samples for each marker. Thirteen polymorphisms have been discovered in selected regions of studied genes, whereas these polymorphisms were only within the GPR177 gene. Eight of these polymorphisms were synonymous and five were in the intron. The tested regions of the ESR1 and RUNX2 genes were monomorphic. The only statistically significant difference was found within the GPR177 gene (exon 2) and the bone length parameter, in the c.443 + 86G > A polymorphism. However, this polymorphism was found in the intron, and no other one was found within the selected regions to show associations with the observed bone parameters.     

Effects of supplemental levels of hesperetin and naringenin on egg quality, serum traits and antioxidant activity of laying hens

Hesperetin and naringenin phytochemicals are naturally occurring flavanoids in citrus fruits. The purpose of this study was to evaluate the effects of supplementing different levels of extracted hesperetin and naringenin on egg quality, serum traits and antioxidant activity in laying hens. Two experiments were conducted, each for 10 weeks, in a completely randomized experiment design. Each had 100 Leghorn laying hens (26 weeks old) randomly assigned into five groups (n = 20) based on dietary categories of hesperetin 0, 0.5, 1, 2, 4 g/kg and naringenin 0, 0.5, 1, 2, 4 g/kg. Experimental results indicated that there was increased (P<0.05) egg production in the 1 g/kg naringenin-supplemented group, but lower (P<0.05) egg production in the hesperetin- and naringenin-supplemented groups given 4 g/kg. Cholesterol content (per gram yolk) and total cholesterol content (per egg) were lower (P<0.05) in the hesperetin- and naringenin-supplemented groups as compared to the control group, and the 2 g/kg hesperetin- and naringenin-supplemented groups showed the most significant difference. Both serum cholesterol and triglyceride concentrations were lower (P<0.05) in the 2 g/kg hesperetin- and naringenin-supplemented groups. The SOD and catalase activities, scavenging O₂⁻ and iron-chelating abilities were higher (P<0.05) in the 2 g/kg hesperetin- and naringenin-supplemented groups, and the trolox equivalent antioxidant capacity was higher (P<0.05) in the 2 g/kg naringenin-supplemented group. The results confirmed that both hesperetin and naringenin could lower serum and egg yolk cholesterol levels, and improve the antioxidant activities, however the measured variables generally showed significant quadratic responses to increasing amounts of the compounds. The recommended supplementation level of hesperetin and naringenin is 2 g/kg of the basal diet for reduced serum and yolk cholesterols contents and increased antioxidant capacity.     

Vitamin e hypervitaminosis in laying hens

The aim of this investigation was to contribute to the problem of overdosing vitamin E. A total of 80 laying hens, divided into 5 groups, were fed diets supplemented with 0; 100, 1000, 10 000 and 20 000 mg/kg vitamin E over a period of 20 weeks within two brooding tests. Laying performance and hatching parameters were registered. All vitamin E doses did not significantly influence health and performances of hens. Vitamin E content of eggs increased from 1 to 4, 21, 46 and 51 mg per egg with vitamin E supplementation. High doses decreased oxidative stability of abdominal fat, vitamin A concentration of liver and egg yolk colour. In both tests vitamin E supplements of 10 000 and 20 000 mg/kg feed resulted in a decrease of living hatched chicken, which demonstrated an adverse effect. Further studies seem to be necessary to explain the effect.     

Influence of high vitamin E dosages on retinol and carotinoid concentration in body tissues and eggs of laying hens

The aim of the study was to contribute to the discussion of overdosing vitamin E in laying hens. A total of 45 laying hens, divided into 5 groups were fed diets supplemented with either 0; 100; 1000; 10 000 or 20 000 mg dl‐α‐tocopheryl acetate/kg diet over a period of 10 weeks. Concentrations of vitamins A and E were measured in plasma, various tissues and egg yolk. Furthermore egg yolk colour and some carotinoids were measured in egg yolks. None of the vitamin E doses significantly influenced performance of the hens. As expected, vitamin E concentration in plasma, all tissue samples and egg yolk was significantly increased with increasing tocopherol content in the diet. The egg yolk showed the highest vitamin E concentration, followed by liver and muscles. Feeding 1000 mg α‐tocopheryl acetate per kg diet resulted in an increase of vitamin A concentration in the liver. Very high doses (10 000 and 20 000 mg/kg diet) significantly decreased retinol concentration in the liver and egg yolk, as well as carotinoid concentration in the egg yolk. The lower carotinoid concentration in egg yolk resulted in a decreased intensity of egg yolk colour. A prooxidative and/or competitive effect of very high doses of vitamin E with other fat soluble substances has been discussed.     

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.     

Dietary curcumin supplementation effects on blood immunological profile and liver enzymatic activity of laying hens after exposure to high temperature conditions

Various environmental factors affect livestock production but heat stress is a major challenge in the poultry farming. Poultry exposes to high temperature alters blood immunological parameters and liver enzymatic function which in turn, suppress the immunity and disease resistance of chickens. Thus, the purpose of present study was to explore the effect of dietary curcumin supplementation on blood immunological biomarker and liver enzymatic activity of laying hens under heat stress conditions. Experimental groups contained two control groups (normal temperature control (NC) and heat stress control (HC) and 3 heat stress curcumin treatment groups (HT100, HT200 and HT300). Hens in HC group with basal diet and heat stress curcumin treatment groups were exposed 6 h/day heat stress (32 ± 1 °C) from 10:00 a.m. to 16:00 p.m. for 9 week. The results of present study showed that heat stressed curcumin treatment group had improved liver weight, WBC values and immunoglobulin level as compared to untreated HC and NC groups. The available results also indicated that laying hens supplemented with curcumin under high temperature conditions had reduced H/L ratio, serum corticosterone levels, inflammatory cytokines response and liver enzymatic activity (ALT) which enhanced the immunity of laying hens under hot climatic conditions. Therefore, it is concluded that curcumin has ability to combat harsh environmental conditions which can be used as anti-inflammatory and immune booster feed additive in the poultry nutrition.     

Selection method and early-life history affect behavioural development, feather pecking and cannibalism in laying hens: A review

The aim of this review is to discuss the effects of selection method and early-life history on the behavioural development of laying hens. Especially in larger groups, laying hens often develop damaging behaviours, such as feather pecking and cannibalism, leading to impaired animal welfare. We hypothesise that the propensity to develop feather pecking and cannibalism is affected by a bird's genetic background and by its early-life history. The genetic background can be influenced by genetic selection. Laying hens are traditionally selected on individual performance, which may lead to co-selection of feather pecking and cannibalism. For hens kept in small groups, it has recently been demonstrated that a novel group selection method, focusing on group performance, can help to reduce cannibalism. However, the biological background behind the success of group selection is unknown. It is also not known whether these results from small groups can be translated to larger groups of laying hens. Regarding early-life history, laying, brooding and rearing conditions have been shown to have major effects on behavioural development and on feather pecking and cannibalism. The presence of a hen during rearing has been shown to improve foraging- and social behaviour, to decrease feather pecking and to decrease fearfulness in chicks. Applying group selection and rearing laying hens in a more natural environment may be key factors in solving the problems caused by feather pecking and cannibalism, especially if the promising results of group selection from small groups in experimental settings can be translated to large-group housing systems.     

A note on fear reaction of three different genetic strains of laying hens to a simulated hawk attack in the hen run of a free-range system

Hens well adapted to free-range systems should react quickly to avoid an attack by a predator but should also be able to return to the hen run soon after to ensure good use of the outside area. To ascertain if some strains cope better with a frightening event the fear reaction of three different genetic strains to a simulated hawk attack in a free-range system was tested. Eight groups of 21 hens composed of the three different strains (LSL, ISA brown and ISA black), were exposed to a simulated hawk attack. Fear reaction and recovery times were video recorded and the use of the hen run was observed. Although the hens did react to the simulated hawk attack as expected by running back into or in direction to the poultry house, no significant strain differences were found either in the reaction or recovery time or in the course of the hen run used. Regarding fear reaction no strain was found to have better features for use as free-range laying hen regarding the behaviour after a frightening event.     

基于盲肠菌群和回肠粘膜基因表达探讨解淀粉芽孢杆菌提高蛋鸡生产性能的潜在机制

【背景】我们的前期工作表明,在蛋鸡饲料中添加解淀粉芽孢杆菌BLCC1-0238可显著提高产蛋性能和蛋品质,但其益生机制还不清楚。【目的】从基因组学和转录组学的视角去揭示益生菌提高蛋鸡生产性能的潜在机制。【方法】利用高通量测序技术比较分析基础日粮组(C组)和解淀粉芽孢杆菌BLCC1-0238添加组(0.06%,2.0×1010 CFU/g,T组)盲肠微生物菌群和回肠粘膜基因表达的差异。【结果】基因组学分析发现,与对照组相比,益生菌添加组的盲肠菌群Chao1和Shannon指数升高;在门水平上,厚壁菌门/拟杆菌门(Firmicutes/Bacteroidetes)的比例显著增加,梭杆菌门(Fusobacteria)和变形菌门(Proteobacteria)的相对丰度降低;在属水平上,厚壁菌门中的考拉杆菌属(Phascolarctobacterium)、乳酸杆菌属(Lactobacillus)、瘤胃球菌科UCG-014属(Ruminococcaceae UCG-014)、厌氧棍状菌属(Anaerotruncus)、瘤胃梭菌属(Ruminiclostridium9)、克里斯滕森氏菌(Christensenellaceae_R-7_group)相对丰度明显增加。转录组学结果表明,回肠粘膜组织中共有356个差异表达基因,其中,上调基因199个,下调基因157个。生物信息学分析提示,上调基因参与了淀粉和蔗糖代谢、半乳糖代谢以及甘氨酸、丝氨酸和苏氨酸代谢等促进物质代谢和营养吸收的过程。【结论】日粮中添加解淀粉芽孢杆菌BLCC1-0238,可通过改善盲肠菌群多样性和肠道营养吸收能力提高蛋鸡的饲料利用率和产蛋性能,具有潜在的实用价值。     

Effect of the combined supplementation of diets with increasing levels of fish and linseed oils on yolk fat composition and sensorial quality of eggs in laying hens

Yolk fatty acid (FA) concentrations and egg quality were evaluated with respect to the inclusion of different levels of marine fish oil (MFO) and linseed oil (LO) in a basal diet. Ten diets were arranged factorially with two levels of MFO addition (15 and 17 g/kg) and five levels of linseed oil (1, 2, 3, 4 and 5 g/kg). Two commercial feeds with no n-3 FA sources added, or containing 15 g MFO/kg served as controls for egg sensorial quality traits. Type of diet did not affect egg production traits, nor total fat content in yolk or saturated and monounsaturated FA concentration in yolk fat. An increase in the inclusion of MFO from 15 to 17 g/kg increased (P<0.05) egg yolk content of total n-3 FA, C_20:5 n-3 and C_22:6 n-3 by 3.3, 9.8 and 3.5%, respectively. Linseed oil addition increased linearly (P<0.001) the level of total n-3 FA and the proportion of C_18:3 n-3, C_20:5 n-3, C_22:5 n-3 and C_22:6 n-3 retained in yolk fat by respectively 131, 12.0, 6.9 and 20.7% between extreme diets. Efficiency of retention of n-3 FA decreased (P<0.001) with dietary level of n-3 FA, and with the dietary ratio of MFO to LO. Both MFO and LO addition decreased (P<0.001) C_20:4 n-6 to C_18:2 n-6 ratio in yolk. Sensorial quality of eggs was not affected by treatments and did not differ with respect of that obtained in commercial n-3 FA enriched eggs. No interaction was found between LO and MFO addition for any of the traits studied. Regression equations have been calculated in order to predict efficiency of retention and yolk fat content of LC n-3 and n-6 FA from dietary characteristics.     

Nest preference of laying hens (Gallus gallus domesticus) and their motivation to exert themselves to gain nest access

In connection with the development of a test method for the attractiveness and appropriateness of nests for laying hens, we carried out an investigation by using a preference test and motivation measuring with the help of push doors. Hens were offered two different nest sites either consisting of a tray filled with wood shavings (litter tray) or a wooden nest box plus wood shavings (nest box). Hens were individually housed in pens (2.0 m × 2.0 m) and had free access to the nest sites until they laid their 15th egg. From that day the hens had to overcome a push door to reach the nest sites. Resistance for entrance was also increased stepwise at the door leading to the hens’ nest. The experiment ended when a hen stopped to lay in her usual nest site for four consecutive days (postexperimental period). The behaviour during the last hour before oviposition was video taped at a level of resistance of 3.5, 6.0, 7.5 and 10.0 N.The hens were categorized into nest and litter layers depending on nest choice. All but one hen pushed maximum resistances between 11.5 and 18.0 N with no differences between nest and litter layers. Behaviour did not significantly change with increasing levels of resistance, but there were significant differences between nest and litter layers. Nest layers spent more time nesting than litter layers while the latter showed a strong tendency to more exploring behaviour. For litter layers, more entries through the push door leading to their nest site and more unsuccessful pushes were detected than for nest layers. According to our results, two types of layers could be distinguished whereas the two were equally motivated to access their nest site.     

Possible risk factors for keel bone damage in organic laying hens

Keel bone damage (KBD) in laying hens is an important welfare problem in both conventional and organic egg production systems. We aimed to identify possible risk factors for KBD in organic hens by analysing cross-sectional data of 107 flocks assessed in eight European countries. Due to partly missing data, the final multiple regression model was based on data from 50 flocks. Keel bone damage included fractures and/or deviations, and was recorded, alongside with other animal based measures, by palpation and visual inspection of at least 50 randomly collected hens per flock between 52 and 73 weeks of age. Management and housing data were obtained by interviews, inspection and by feed analysis. Keel bone damage flock prevalences ranged from 3% to 88%. Compiled on the basis of literature and practical experience, 26 potential associative factors of KBD went into an univariable selection by Spearman correlation analysis or Mann–Whitney U test (with P<0.1 level). The resulting nine factors were presented to stepwise forward linear regression modelling. Aviary v. floor systems, absence of natural daylight in the hen house, a higher proportion of underweight birds, as well as a higher laying performance were found to be significantly associated with a higher percentage of hens with KBD. The final model explained 32% of the variation in KBD between farms. The moderate explanatory value of the model underlines the multifactorial nature of KBD. Based on the results increased attention should be paid to an adequate housing design and lighting that allows the birds easy orientation and safe manoeuvring in the system. Furthermore, feeding management should aim at sufficient bird live weights that fulfil breeder weight standards. In order to achieve a better understanding of the relationships between laying performance, feed management and KBD further investigations are needed.     

Development of locomotion over inclined surfaces in laying hens

The purpose of the present study was to evaluate locomotor strategies during development in domestic chickens (Gallus gallus domesticus); we were motivated, in part, by current efforts to improve the design of housing systems for laying hens which aim to reduce injury and over-exertion. Using four strains of laying hens (Lohmann Brown, Lohmann LSL lite, Dekalb White and Hyline Brown) throughout this longitudinal study, we investigated their locomotor style and climbing capacity in relation to the degree (0 to 70°) of incline, age (2 to 36 weeks) and the surface substrate (sandpaper or wire grid). Chicks and adult fowl performed only walking behavior to climb inclines ⩽40° and performed a combination of wing-assisted incline running (WAIR) or aerial ascent on steeper inclines. Fewer birds used their wings to aid their hind limbs when climbing 50° inclines on wire grid surface compared with sandpaper. The steepness of angle achieved during WAIR and the tendency to fly instead of using WAIR increased with increasing age and experience. White-feathered strains performed more wing-associated locomotor behavior compared with brown-feathered strains. A subset of birds was never able to climb incline angles >40° even when using WAIR. Therefore, we suggest that inclines of up to 40° should be provided for hens in three-dimensional housing systems, which are easily negotiated (without wing use) by chicks and adult fowl.     

Influence of physical activity on stability of somatotypes in boys.

The purpose of this study is to examine the stability of somatotypes of boys with different activity levels followed longitudinally for eight years. Thirty-nine boys ated 11 were somatotyped annually (until 18 years) using the Heath-Carter anthropometric rating method. Three sub-groups were formed according to their levels of systematic physical activity designated High, Moderate, and Low Activity. There were no differences in somatochart distributions of the Total Group, but there were considerable individual changes which cancelled each other in group comparisons. All boys changed their somatotype ratings at least once and 67% changed in component dominance. The inter-age correlations for the components were generally of poor prediction value, and the somatotype migratory distances showed considerable variation. The Total Group means differed on three components and this was attributed to differences within the Moderate Activity Group, the only sub-groups to differ. It was concluded that changes occur in somatotypes of adolescent boys, but the influence of physical activity on the changes is inconclusive.     

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.     

Oviduct-specific expression of tissue plasminogen activator in laying hens

Egg-laying hens are important candidate bioreactors for pharmaceutical protein production because of the amenability of their eggs for protein expression. In this study, we constructed an oviduct-specific vector containing tissue plasminogen activator (tPA) protein and green fluorescent protein (pL-2.8OVtPAGFP) and assessed its expression in vitro and in vivo. Oviduct epithelial and 3T3 cells were cultured and transfected with pL-2.8OVtPAGFP and pEGP-N1 (control vector), respectively. The pL-2.8OVtPAGFP vector was administered to laying hens via a wing vein and their eggs and tissues were examined for tPA expression. The oviduct-specific vector pL-2.8OVtPAGFP was expressed only in oviduct epithelial cells whereas pEGP-N1 was detected in oviduct epithelial and 3T3 cells. Western blotting detected a 89 kDa band corresponding to tPA in egg white and oviduct epithelial cells, thus confirming expression of the protein. The amount of tPAGFP in eggs ranged 9 to 41 ng/mL on the third day after vector injection. The tPA expressed in egg white and oviduct epithelial cells showed fibrinolytic activity, indicating that the protein was expressed in active form. GFP was observed only in oviducts, with no detection in heart, muscle, liver and intestine. This is the first study to report the expression of tPA in egg white and oviduct epithelial cells using an oviduct-specific vector.     

The presence of extreme feather peckers in groups of laying hens

Feather pecking is a serious economic and welfare problem in laying hens. Feather damage occurs mainly through severe feather pecking (SFP). Selection experiments have proved that this behavior is heritable and lines have been divergently selected for high (HFP) and low feather pecking (LFP). The number of bouts of SFP per hen follows a Poisson distribution with a maximum nearby 0. A few studies indicate that the distribution within flocks is not homogenous but contains sub-groups of birds showing extremely high levels of feather pecking (EFP). It was the aim of the current study to re-analyze data on SFP of lines selected for HFP/LFP and their F2 cross so as to uncover hidden sub-populations of EFP birds. Data of seven selection generations of HFP and LFP selection lines as well as their F2 cross have been used. We fitted a two-component mixture of Poisson distributions in order to separate the sub-group of EFP from the remaining birds. HFP and LFP lines differed mainly in mean bouts per bird. The proportion of EFP was only marginal in the LFP as compared with the HFP and the F2 population. Selection for LFP did not result in total elimination of EFP. The presence of even small proportions of EFP may play an important role in initiating outbreaks of feather pecking in large flocks. Further studies on feather pecking should pay special attention to the occurrence of EFP sub-groups.     

Modulation of energy and protein supplies in sequential feeding in laying hens

Sequential feeding (SF) consists of splitting energy (E) and protein/calcium (P) fractions temporally, improving the feed conversion ratio (FCR) of hens compared with a continuous distribution during the day. In a previous study, the E fraction (with a low level of protein) was provided in the morning, whereas the P fraction (with low level of energy) was given in the afternoon. However, there is no clear evidence that a requirement in energy or proteins is connected to these distribution sequences, whereas the requirement for calcium is known to be required in the afternoon. To evaluate the effects on performances of the modulation of energy and protein supplies in SF, five different sequential treatments were offered: E0P0/E0P0; E+P+/E−P−; E+P−/E−P+; E0P+/E0P− and E+P0/E−P0 where E+ represents a high energy level, E0 a moderate one and E− a low one (with the same meaning for P regarding protein supply). Afternoon fractions were provided with particulate calcium. A total of 168 Hendrix hens were housed in individual cages from 20 to 39 weeks of age in two environmentally contrasted rooms. Feed intake in the morning and afternoon fractions, egg production, egg weight, BW and weight of digestive organs were recorded. No diet effect was observed concerning feed intake, egg production and BW. These results suggested that hens are not able to fit their feed intake on energy or protein level of fractions within half-day duration, whereas at the day scale same protein and energy intakes were observed. Moreover, the time of nutrient distribution in feeding did not seem to have an impact on birds’ performances. These studies have also demonstrated that, despite strong environmental pressure, the hens with SF had attenuated performance but continue to produce eggs.