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Among land vertebrates, the laying hen stands out due to its great reproductive efficiency: producing an egg daily all year long. This production rate makes the laying hen a special model animal to study the general process of reproduction and aging. One unique aspect of hens is their ability to undergo reproductive plasticity and to rejuvenate their reproductive tract during molting, a standard industrial feed restriction protocol for transiently pausing reproduction, followed by improved laying efficiency almost to peak production. Here we use longitudinal metabolomics, immunology, and physiological assays to show that molting promotes reproduction, compresses morbidity, and restores youthfulness when applied to old hens. We identified circulating metabolic biomarkers that quantitatively predict the reproduction and age of individuals. Lastly, we introduce metabolic noise, a robust, unitless, and quantifiable measure for heterogeneity of the complete metabolome as a general marker that can indicate the rate of aging of a population. Indeed, metabolic noise increased with age in control hens, whereas molted hens exhibited reduced noise following molting, indicating systemic rejuvenation. Our results suggest that metabolic noise can be used as a quick and universal proxy for assessing successful aging treatments, accelerating the timeline for drug development., (© 2023. The Author(s).)

In this study, the effects of cage type and the light-dark cycle on the behavior of hens before, during, and after forced molting were determined. For this purpose, 73-week-old hens were placed in the two different cage types used in the experiment. The barley method was used to induce molting at 75 weeks of age. The molting period lasted 35 days in total. The frequencies of walking, feeding, comforting, and preening behaviors were affected by the forced molting period. The addition of enrichment materials to the cage did not cause any changes in hen behavior. The light-dark cycle was important for the frequency of all examined behaviors except fighting behavior. No interaction was found among forced molting period, cage type, and light-dark cycle regarding behaviors. It is thought that there is a decrease in welfare-related behaviors at the end of the laying period and this decrease does not increase with forced molting using barley; on the contrary, appropriate forced molting applied to hens during this period will increase welfare. It was observed that cage type did not cause a significant difference in hen behavior during the forced molting period., (© 2024 The Author(s). Animal Science Journal published by John Wiley & Sons Australia, Ltd on behalf of Japanese Society of Animal Science.)

1. Moulting is a natural physiological process in birds when they shed their old feathers and replace them with new ones, and it is followed by reproductive quiescence resulting in reduced egg production. Different birds undergo moulting at different points in their life. Some birds have seasonal moulting while some moult at the end of their breeding cycle. This review will mainly focus on moulting practices associated with commercial layer birds because, in all other bird types, this is not managed.2. Commercial farms commonly analyse the cost-benefit ratio to decide the time and method to adopt for moulting. Commercial layer farms adopt different practices to force birds out of moult and restart the production cycle. Animal welfare groups consider this as stressful and against animal welfare, raising questions about the ethics of this practice.3. Many studies have been conducted using complete or partial feed withdrawal and non-feed withdrawal programs to measure their effectiveness in maintaining animal welfare, economy, and post-moult performance in mind.4. Animal welfare should not be compromised during moulting. The United States Egg Producers and other such groups from the United Kingdom and Europe have decided to sell eggs produced only through a non-feed withdrawal moulting programs.

How dermis maintains tissue homeostasis in cyclic growth and wounding is a fundamental unsolved question. Here, we study how dermal components of feather follicles undergo physiological (molting) and plucking injury-induced regeneration in chickens. Proliferation analyses reveal quiescent, transient-amplifying (TA) and long-term label-retaining dermal cell (LRDC) states. During the growth phase, LRDCs are activated to make new dermal components with distinct cellular flows. Dermal TA cells, enriched in the proximal follicle, generate both peripheral pulp, which extends distally to expand the epithelial-mesenchymal interactive interface for barb patterning, and central pulp, which provides nutrition. Entering the resting phase, LRDCs, accompanying collar bulge epidermal label-retaining cells, descend to the apical dermal papilla. In the next cycle, these apical dermal papilla LRDCs are re-activated to become new pulp progenitor TA cells. In the growth phase, lower dermal sheath can generate dermal papilla and pulp. Transcriptome analyses identify marker genes and highlight molecular signaling associated with dermal specification. We compare the cyclic topological changes with those of the hair follicle, a convergently evolved follicle configuration. This work presents a model for analyzing homeostasis and tissue remodeling of mesenchymal progenitors., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2021. Published by The Company of Biologists Ltd.)

The purpose of the present study was to determine the histological and ultrastructural changes in the luminal epithelium of the shell gland associated with natural moulting. Samples of the shell gland from laying (32 weeks old) and moulting (75 weeks old) hens were studied using histological, histochemical and electron microscopic techniques. In addition, TUNEL was used to demonstrate the distribution of apoptotic cells in the luminal epithelium of the shell gland. Autophagy, characterized by the presence of autophagosomes and autolysosomes, was evident in the early stages of degeneration in non-ciliated, ciliated and mitochondrial cells. The intermediate and advanced stages of regression in non-ciliated as well as mitochondrial cells occurred via apoptosis, while both apoptotic and necrotic ciliated cells were observed during the later stages of degeneration. The results of the present study suggest that a synergy of autophagy, apoptosis and necrosis is involved in the involution of the shell gland during natural moulting., (© 2020 Blackwell Verlag GmbH.)

This study examined the effect of water-chilled perches on hen production and physiological responses to induced molt during elevated temperatures. A total of 288White Leghorns at 82 wk of age were housed in 36 cages of 6 banks. Each bank was assigned to 1 of 3 treatments: cooled perches, air perches, and no perches. The hens were subjected to 2 heat episodes during their first laying cycle at week 21 to 35 and week 73 to 80, respectively. The hens were subjected to a 28 D nonfasted molting regimen starting at 85 wk of age. Cyclic heat of 32°C (6:00 am to 6:00 pm) was applied daily during the molting period. After molt, hens were returned to a layer diet and housed under thermoneutral condition. Two birds per cage were monitored for BW change during molt. Egg production was recorded daily. Feed utilization was measured during molt at 86 and 88 wk of age. Egg weight and eggshell traits were examined at 84 wk (pre-molt) and post-molt at 92, 96, and 104 wk of age. Rectal temperature and blood samples were collected from 2 birds per cage at the end of molt. Blood samples were used for determining heterophil/lymphocyte ratio, corticosterone, and thyroid hormones. Plumage condition was examined at 22 wk post-molt. Compared to control and air perch hens, cooled perch hens had higher feed usage and greater BW loss, lower heterophil/lymphocyte ratios (P < 0.05) with no difference in thyroid hormones and corticosterone at the end of molt. Cooled perch hens also had higher egg production beginning at 98 wk of age (Ptreatment*age < 0.0001) than control hens and sometimes the air-perch hens. Cooled perch hens had higher rectal temperature than control but not air perch hens at end of molt. Moreover, cooled perch hens had better breast feather scores than air perch hens but worse vent plumage (P ≤ 0.05) than both control and air perch hens. These results indicate that the provision of cooled perches assists hens with better adaptation to stressors, such as induced molt plus heat exposure, resulting in improved post-molt egg production., (Published by Oxford University Press on behalf of Poultry Science Association 2019.)

The molting cycle of feathers includes an anagen (growth) stage, a likely catagen stage where the feather follicles degenerate, and a resting stage where fully grown feathers remain in their follicles and are functional before molting. However, the cytological changes involved in the resting and molting stages are poorly known, so the results of an ultrastructural analysis of these processes in adult chick feathers are presented here. The study showed that the dermal papilla shrinks, and numerous cells present increased heterochromatin and free collagen fibrils in the extracellular matrix. Degeneration of the germinal epithelium of the follicle-the papillary collar-occurs with an initial substantial contraction of cells followed by an increase in heterochromatin, vesicle and lipid accumulation, and membrane and organelle degeneration. Desmosomes are still present between degenerating epithelial cells, but ribosomes and tonofilaments disappear. This suggests that cell necrosis initially proceeds as a major contraction resembling apoptosis-a process termed necroptosis, which was previously also shown to occur during the formation of barbs and barbules in mature down and pennaceous feathers. This study suggests that, aside from apoptosis, the collar epithelium degenerates due to external factors, in particular the retraction of blood vessels supplying the dermal papilla. In contrast, revascularization of the dermal papilla triggers a new phase of feather growth (anagen).

The aim of this study was to investigate the efficacy of feed-grade preparations of mannan oligosaccharides ( MOS: ) and oregano essential oil ( OEO: ) in forced molted or fully fed 82-week-old, laying hens. A 2 × 3 factorial experiment investigated the influence of molting vs. full feeding and dietary supplements [i.e., unsupplemented control, MOS (1 g/kg) diet, and OEO (24 mg/kg) diet] on production parameters, egg quality, serum stress indicators, blood constituents, tibial characteristics, liver antioxidant status, and cecal microflora composition. A total of 864 Single Comb White Leghorn hens were randomly assigned to 6 treatments, each with 6 replicates of 24 hens each, and studied for 25 wk. Hens were fed a molt diet containing of 50% alfalfa and 50% wheat bran ( AA+WB: ) for 12 d, then returned to the laying ration. Results indicate that molt vs. full feed impacted more on most variables measured than supplementation or supplement type. Significant (P < 0.01) interactions between molting and diet were observed for the egg production, egg weight, egg mass, and feed conversion ratio ( FCR: ). In fully fed hens, MOS supplementation improved (P < 0.01) the egg production, egg weight, and FCR, and an OEO addition significantly improved the egg production and FCR in forced molted hens. Molting improved egg quality despite the significant regression in ovary and oviduct weight (P < 0.01), though supplements showed no influence. The bone ash (P < 0.01) and mineral content (P < 0.05) of molted hens were significantly lower than those of fully fed counterparts; however, poor mineralization was not reflected in the bones' mechanical properties. No significant differences were observed among treatments for hematological characteristics. Both the MOS and particularly the OEO supplementation improved (P < 0.01) liver antioxidant status and mitigated the significant increase in cecal pathogenic bacteria after molt. Our results indicate that full feeding with an aa+wb diet is an effective non-feed-removal method for molted hens, the benefit of which can be improved with MOS and OEO supplementation., (© 2016 Poultry Science Association Inc.)

Apolipoprotein D (APOD) is a glycoprotein which is widely expressed in mammalian tissues. It is structurally and functionally similar to the lipocalins which are multiple lipid-binding proteins that transport hydrophobic ligands and other small hydrophobic molecules, including cholesterol and several steroid hormones. Although multiple functions for APOD in various tissues have been reported, its expression, biological function, and hormonal regulation in the female reproductive system are not known. Thus, in this study, we focused on correlations between APOD and estrogen during development, differentiation, regression, and regeneration of the oviduct in chickens and in the development of ovarian carcinogenesis in laying hens. Results of the present study indicated that APOD messenger RNA (mRNA) expression increased (P < 0.001) in the luminal and glandular (GE) epithelia of the chicken oviduct in response to diethylstilbestrol (a nonsteroidal synthetic estrogen). In addition, the expression of APOD mRNA and protein decreased (P < 0.001) as the oviduct regressed during induced molting, and gradually increased (P < 0.001) with abundant expression in GE of the oviduct during recrudescence after molting. Furthermore, APOD mRNA and protein were predominantly localized in GE of cancerous, but not normal ovaries from laying hens. Collectively, results of the present study suggest that APOD is a novel estrogen-stimulated gene in the chicken oviduct which likely regulates growth, differentiation, and remodeling of the oviduct during oviposition cycles. Moreover, up-regulated expression of APOD in epithelial cell-derived ovarian cancerous tissue suggests that it could be a candidate biomarker for early detection and treatment of ovarian cancer in laying hens and in women., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Cathepsins (CTSs) are peptidases that have biological roles in degrading extracellular matrix, catabolism of intracellular proteins, and processing of pro-hormones. Of these, cathepsin L (CTSL) is closely associated with morphological changes in reproductive organs required for proper function in mammals, including humans and mice, but little is known about CTSL in avian species. In the present study, the expression of CTSL was investigated in the oviduct of hens during regression and recrudescence in response to molting. Our results revealed that expression of CTSL mRNA increased (P<0.001) when the oviduct underwent regression during the molting period in hens. In situ hybridization and immunohistochemial analyses detected CTSL mRNA and protein predominantly in the luminal (LE) and glandular epithelia (GE) during regression of the oviduct, but not during regeneration of the oviduct. Expression of CTSL decreased in the oviduct of chicks treated with diethylstilbestrol (DES, a synthetic estrogen agonist). Furthermore, we discovered four miRNAs including miR-23b, miR-551, miR-1464 and miR-1803 that regulate expression of the CTSL gene at the post-transcriptional level, which suggests that CTSL mRNA can be regulated by specific miRNAs via 3'-UTR in chickens. Results of the present research suggest that estrogen regulates expression of CTSL during regression of the oviduct during molting and that down-regulation of CTSL is likely a prerequisite for the normal regeneration of oviductal tissues following molting in laying hens., (Copyright © 2014 Elsevier Inc. All rights reserved.)

The purpose of this study was to investigate the effect of dietary vitamin E and vitamin C, probiotics mixture and protein level and their combination on semen quality and immunohistochemical study of some pituitary hormones in male broiler breeders. One hundred and eighty male broiler breeders 65 weeks old were divided into six groups by completely randomized design. The birds were subjected to zinc-induced molt by mixing zinc oxide at the rate of 3000mg/kg in the feed. After molting, one group was fed control diet (CP16%). The other groups were fed vitamin E (100IU/kg), vitamin C (500IU/kg), probiotics (50mg/L of drinking water), protein (CP14%) and combination of these components. These treatments were given for five weeks. After the feeding period, semen samples were taken and analyzed for semen volume, sperm concentration, motility and dead sperm percentage. Pituitary samples were collected from three birds per replicate and were processed for immunohistochemical study. The results of semen quality parameters revealed that semen volume and sperm motility were significantly high in the vitamin E fed group, while the dead sperm percentage decreased significantly in the vitamin C group. The morphometric analysis revealed that compared to other groups, vitamin E caused a significant increase in the size and area of FSH, LH gonadotropes and lactotropes. These results showed that vitamin E alone may play some role in the enhancement of semen quality and growth of gonadotropes and lactotropes., (Copyright © 2013 Elsevier GmbH. All rights reserved.)

In the present study, the expression profile of luteinizing hormone receptor (LHR) was investigated in the ovary, magnum and uterus and in hierarchcal follicles (F-1, F-2, F-3 and F-4) of hens subjected to moulting to establish their involvement in moulting and presence in non-gonadal tissues. Fifty-two layers (72 weeks) were subjected to moult for a period of 14 days. Four birds were sacrificed each time on 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12 and 14 days of moulting, and samples (ovary, magnum, uterus and hierarchal follicles) were collected aseptically for the quantitative study by real-time polymerase chain reaction. The ovary, isthmus, uterus and magnum weight reduced significantly during induced moulting. From the 4 DOM, this reduction was drastic and reached approximately 80% of original weight in the case of ovary, isthmus and magnum and approximately 65% of original weight in the case of uterus on 14 DOM. Ovarian yellow follicles decreased gradually from 1 DOM to 4 DOM, after that no normal yellow follicle was observed in moulted bird. The number of atretic follicles increased gradually during the course of induced moulting, reaching the peak at 5 DOM. The LHR mRNA was detected in non-gonadal tissues like magnum and uterus. The LHR expression was significantly (p < 0.05) down regulated in ovary, magnum and uterus throughout the treatment. These results indicated that LHR may have a role in reproductive tissue regression during moulting., (© 2012 Blackwell Verlag GmbH.)

Tight junctions in the mucosal epithelium have essential roles as a mucosal barrier to prevent invasion of microbes into the hen oviduct tissue. The aim of this study was to determine the effects of the egg-laying phase and estradiol on the expression of tight junction molecule "claudins" in the lower oviductal segments in hens. White Leghorn laying and molting hens were used. Molting hens were given either sesame oil (vehicle) or estradiol benzoate (N = 5 per group) via injection. The lower segments of oviduct (isthmus, uterus, and vagina) of these birds were collected. Gene expression of claudin-1, -3, -5, lipopolysaccharide-induced TNFα factor (LITAF), and IFN(γ) was analyzed by quantitative reverse transcription polymerase chain reaction, and localization of claudin-1 was examined by immunohistochemistry. Permeability in the mucosal epithelium was assessed by intrauterine injection of fluorescein isothiocyanate-dextran. Expression of claudin-1, -3, and -5 genes and density of claudin-1 protein in the lower oviductal segments were higher in laying hens than in molting hens (P < 0.01); their expression was upregulated by estradiol (P < 0.01). Expression of LITAF and IFN(γ) genes was higher in molting hens than in laying hens. More fluorescein isothiocyanate-dextran infiltrated into the intercellular space of the uterus mucosal epithelium in molting hens than in laying hens and estradiol-treated molting hens. In conclusion, we inferred that barrier functions of the mucosal epithelium in the lower oviductal segments might be disrupted because of reduced claudin expression in molting hens, which might increase the susceptibility of mucosal tissue during the molting phase., (Copyright © 2013 Elsevier Inc. All rights reserved.)

The study aimed to investigate the comparative effect of vitamins E and C, probiotics, lower than normal protein level and the combination of these treatments in male broiler breeders after Zn-induced molting. One hundred and eighty birds at 65 weeks of age were divided into six groups (five replicates per group) in a completely randomized block design. Birds were induced to molt by mixing zinc oxide in feed at the rate of 3000 mg/kg of feed. Upon completion of molting, birds were fed different supplements. One group was kept as a control, while the other groups were fed: vitamin E (100 IU/kg), vitamin C (500 IU/kg), probiotics (50 mg/L of water), lower dietary protein level (14%) or the combination of these treatments. Results revealed that vitamins (E and C) supplementation increased (P<0.05) semen volume, however, sperm motility and egg fertility were significantly higher only in vitamin E supplemented group compared to control. The dead sperm percentage decreased (P<0.05) in vitamin C fed group, whereas, sperm concentration did not differ (P>0.05) between the control and treated groups. The results validate the key role of antioxidant vitamins in male broiler breeders' fertility after Zn-induced molting., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Dietary zinc was used to induce molt in 200 White Leghorn birds in caged housing at the age of 70 wk. The birds were equally and randomly allocated to 4 groups each of 50 birds as G1 (control; CP 16%, no supplement), G2 (CP 18%, no other supplement), G3 (CP 16%, symbiotic Perfectin: 85 mg•L(-1) in drinking water daily), and G4 (CP 16%, probiotic Protexin: 85 mg•L(-1) in drinking water daily) after the completion of molt. The sampling was conducted 3 times at 5% production, peak production, and end of production with 15 birds being killed at each sampling from each group to collect the blood for harvesting of serum. Serum health markers including total oxidant status, total antioxidant capacity, homocysteine concentration, and paraoxonase, arylesterase, and ceruloplasmin activity were determined by the prescribed assays. The overall total antioxidant capacity was increased, whereas total oxidant status and homocysteine concentrations were reduced significantly (P ≤ 0.01) in all the supplemented groups compared with the control. The paraoxonase and ceruloplasmin activity were enhanced (P ≤ 0.01) in the supplemented groups compared with the control, and arylesterase activity was increased (P ≤ 0.01) in only G2 compared with the other groups. Although protein supplementation significantly reduced the oxidative stress, supplementation with symbiotic and probiotic also improved the health status by decreasing the oxidative stress in the birds.

The aim of this study was to determine the differences in the mucin expression that forms a mucosal surface barrier in the oviduct between laying and molting hens. The lower segments of oviducts (isthmus, uterus, and vagina) of White Leghorn laying and molting hens were collected. Localization and gene expression of mucosal mucin were analyzed by quantitative reverse-transcription PCR of mucin mRNA and mucin5AC immunohistochemistry. Sugar residues were localized by lectin (WGA or Jacalin) histochemistry. Expression of mucin mRNA was significantly declined in the lower oviductal segments in molting compared with laying hens. Immunoreactive-mucin5AC was localized in the mucosal epithelium and on the epithelial surface of laying hens, whereas it was reduced in molting hens. Substances positively stained by WGA and Jacalin were identified on the surface of the mucosal epithelium in the lower oviductal segments in laying and molting hens. These results suggest that mucin synthesis in the lower segments of the oviduct is reduced, although the existence of WGA- and Jacalin-positive sugars may be kept even in the molting phase. The reduction of mucin synthesis may result in a decline of mucosal barrier function in the molting phase.

The purpose of this study was to evaluate alternative molting protocols assessing hen welfare and performance during and after molt. Hyline W-36 pullets were housed at 15 wk of age, and their egg production was obtained during this first cycle. When birds were 80 wk, the following molting treatments were applied: a conventional molt consisting of 10 d of fasting followed by cracked corn for 8 d and a pullet developer diet for 10 d; and 4 alternative molting programs: a soy hulls-based diet (12% CP, 1,455 kcal/kg of ME, and 1.38% Ca) offered for 14 d followed by cracked corn for 4 d and a pullet developer diet for 10 d; and the other 3 molt regimens consisted of feeding soy hulls for 4, 8, or 12 d followed by 10, 6, or 2 d, respectively, of a soy hulls-based diet and 4 d of cracked corn plus 10 d of a pullet developer diet. A nonmolted group of birds was fed a laying hen diet during the experimental period. Hen-day egg number was recorded daily for 56 wk (through 80-136 wk of age). The nonmolted hens showed lower hen-day production and fewer intact eggs and a higher number of cracked and shell-less eggs compared with those of the molted hens (P < 0.0001). A significant treatment by age effect (P < 0.0001) was observed for the variables of high-density lipoproteins and triglycerides. Control hens showed the lowest high-density lipoprotein concentration and the highest triglyceride levels at 84 wk of age when compared with all treatments. The lack of difference in heterophil-to-lymphocyte ratio during molt suggests little influence of molting protocols on this variable. Regardless of the treatments, molting was deleterious to bone quality. A high mobilization of Ca through bone resorption for eggshell formation could explain the results obtained. Behavioral patterns coincided with a decline in frustration activities and an increase in alertness as molt proceeded until 83 wk of age. Alternative molting diets consisting of soybean hulls were successful in providing acceptable postmolt egg production performance.

The objectives of this study were to compare the behaviors, postures, and heterophil-to-lymphocyte ratios (H:L) of laying hens housed in a cage system when offered a Ca premolt treatment and low-energy molt diets vs. a traditional feed withdrawal (FW) treatment during and after molt. A total of 144 Hy-Line W-36 hens (85 wk of age), housed 3 hens/cage (413 cm(2)/hen), were used. Hens were allotted to treatments according to a randomized complete block design, with the cage location and initial BW as the blocking criteria. Six treatments were compared in a 2 × 3 factorial arrangement with 2 Ca premolt treatments (fine or coarse) and 3 low-energy molt diets (FW, soybean hulls, or wheat middlings). The 2 Ca premolt treatments differed only in Ca particle size (fine was 0.14 mm and coarse was 2.27 mm mean diameter). Two postures and 5 behaviors were recorded and H:L was measured. Data were analyzed using the MIXED procedure of SAS, with P < 0.05 considered significant. There were no differences in behaviors, postures, or H:L during the premolt baseline period. The Ca premolt treatment had no carryover effects during or after molt for behaviors or postures. During molt, hens in the FW treatment were more active, and they ate and drank less compared with hens fed soybean hulls or wheat middlings, but there were no differences in aggression, nonnutritive pecking, or sitting. Drinking and aggression during and after molt were not different, but hens postmolt engaged in more sitting and feeding and less activity, nonnutritive pecking, and preening compared with during molt. There were no differences in H:L during or after molt. In conclusion, a Ca premolt treatment did not affect the behavior of the laying hen. The low-energy molt diets did not adversely affect behavior compared with FW and did not increase H:L; therefore, they could be useful alternatives for inducing molt in laying hens.

The objective of this study was to determine the distribution of somatotrophs and lactotrophs and conduct a morphometrical analysis of immunoreactive somatotrophs and lactotrophs in the pituitary glands of White Leghorn Hens (Gallus domesticus) during the period of induced moult. We divided the periods of induced moulting into three phases viz. 7, 14 and 21 days. The labeled alkaline-phsphatase method with anti-GH (growth hormone) and anti-PRL (prolactin) as a primary antibody was used to detect somatotrophs and lactotrophs, in the midsagital sections of chicken adenohypophysis. Immunohistochemistry showed that somatotrophs are not only confined to the cephalo-caudal axis but can also be found in the caudal lobe; while lactotrophs were distributed in both lobes of the anterior pituitary gland at all stages of moulting (7, 14 and 21 days). Lactotrophs were of different shapes but somatotrophs were oval to round in morphology. At the given stages of induced moulting, some hypertrophied lactotrophs were also present after 7 days of induced moult in the anterior pituitary gland. However, there were moulting-related changes: from 7 to 21 days of induced moulting the immunoreactive-PRL cell population decreased, while the mean lactotroph size was more than that of somatotrophs. Basic quantitative and morphological information relating to somatotrophs and lactotrophs during the period of induced moult in laying hens is reported here and the changes brought about by induced moulting are restricted to PRL positive cells rather than GH positive cells.

1. The applicability of different ratios of bitter vetch seed as a new method for moult induction in laying hens was studied. The effectiveness of bitter vetch seed on post-moult production and post-moult egg quality was also investigated. 2. A total of 120 Single Comb White Leghorn hens, 78 weeks of age, were used in this study. The hens were randomly assigned to 5 treatment groups of 24 birds each. The treatments were 30% bitter vetch seed (BV30) diet, 60% bitter vetch seed (BV60) diet, 90% bitter vetch seed (90BV) diet, feed withdrawal method (FW) and full-fed non-moulted control (CON). 3. Egg production ceased first in FW and BV90 treated hens and last in BV30 treated hens. As the percentage of bitter vetch seed increased in the moulting ration, feed intake decreased and body weight loss increased during the 10-d moult induction period. Time to first egg production was significantly greater in hens exposed to the FW and BV90 diets. 4. FW and BV90 treatment hens had significantly higher hen-d egg production than non-moulted control hens. Egg weight was significantly higher in BV30 and BV90 treatments. There were no differences in egg mass, feed intake and mortality among experimental treatments during the post-moult period. 5. No significant improvements were observed in exterior or interior egg quality in moulted hens, except for Haugh units, which were significantly higher in moulted hens when compared to the non-moulted control hens. 6. In conclusion, the present study showed ad libitum feeding of a layer ration with 90% of bitter vetch seed for 10 d proved to be effective for inducing moult, increasing post-moult egg production and improving some internal egg quality parameters.

A number of 252, 70 week old, single comb white leghorn hens at the end of their first production cycle were divided into two groups: group-I; molted by fasting and group-II; by high dietary zinc to obtain the second and third production cycle. Sampling was carried out at 5%, peak and end of the second production cycle and at 5% and peak of the third production cycle. Serum zinc decreased at the peak of the second production cycle, while increased at the 5% of the third production cycle in zinc-molted group. Serum copper and manganese increased at the 5% and peak of the second production cycle in zinc-molted group. Egg albumin zinc and copper increased at the 5% and peak of the second as well as third production cycles in zinc-molted group. Egg albumin manganese increased at 5% during the second production cycle in fasting molted group. Egg yolk zinc increased at the peak of the second production cycle in zinc-molted group. Egg yolk copper increased at the 5% of second production cycle but decreased during the third production cycle in zinc-molted group. Egg yolk manganese increased at the 5% and at the end of the second production cycle in zinc molted group. Egg yolk manganese and iron decreased during the third production cycle in zinc-molted group. Eggshell zinc decreased at the end of the second production cycle in the zinc-molted group, while eggshell copper increased at the peak of the second as well as third production cycles in fasting molted group.

The purpose of the present study was to investigate egg production performance, immunohistochemical (IHC) identification, localization and morphological characterization of moulted hen's pars distalis luteinizing hormone (LH) gonadotrophs. This system has been validated for chicken LH by immunodetection of purified hormones on dot blots, western blotting and by specific labelling of cells on sagittal sections of chicken pituitaries. This study showed that egg production and egg weight increased significantly after zinc-induced moult when compared with fasting-induced moult. Luteinizing hormone-gonadotrophs are of numerous shapes and distributed throughout the adenohypophysis, with densely populated regions on the margins of pars distalis. Moulting of laying hens was associated with a significant increase in immunoreactive LH gonadotroph cell count and cell size, which peaked at the subsequent production stages. Zinc-induced moulting also caused a significant increase in LH gonadotroph cell size as compared with fasting-induced moulting. The nuclear size of LH-positive cells was distinct during decreased egg production. Our results validate the use of ovine LHbeta antibodies for the IHC localization of chicken LH gonadotrophs. This technology was further applied to the characterization of the shape and morphological changes of gonadotrophs as a function of the egg production status of the birds.

The objective of this study was to determine if increasing hen age and 3 different molting treatments influenced the total microflora counts or the prevalence of Salmonella spp. on the exterior of the egg shell, within the interior shell, or in the contents. Eggs from Hy-Line W-98 and Bovans White layer strains were sampled approximately every 28 d from 70 to 114 wk of age, with the molting period from 66 to 70 wk of age. Layers were utilized from the 35th North Carolina Layer Performance and Management Test and managed under identical husbandry practices. This study consisted of nonfasted, nonmolted, and feed-restricted treatments with the use of 135 eggs per layer strain, for a total of 270 eggs sampled per period. The exterior, interior shell, and contents were spiral plated onto plate count agar to calculate the total aerobic counts. Additional preenrichment, enrichment, conformational, and biochemical procedures were performed to test for the presence of Salmonella spp. Hen age and molting treatment significantly (P < 0.05) affected the microbial loads on all 3 egg components. Exterior, interior, yolk, and albumen counts increased during the molt period to as much as 1 log unit higher than the highest countable plate, which was 10(5). Exterior, interior, and contents counts significantly increased (P < 0.05) during period 15, with a significant increase (P < 0.05) in the interior also in period 14, and in the contents in periods 14 and 17. There were a total of 360 egg pools, and of those, 4 were positive Salmonella samples. Both the interior and exterior shell components and 2 of the 3 molting treatments had positive samples. Of these positives, 4 were confirmed as Salmonella Braenderup. Three positives were associated with the interior component, whereas 1 positive was associated with the exterior shell component. Three of the 4 samples were related to the nonfasted treatment, whereas the remaining positive was found in the non-molted treatment.

Non-feed removal molting programme in commercial brown laying hens and its influence on pre-molting, post-molting and end of cycle egg quality traits were investigated. Overall 54 birds were randomly divided into three treatment groups and each group was fed with one of the following diets during 10 days of molting period: (i) grain barley, (ii) alfalfa meal, or (iii) commercial layer ration (non-molted control group). Eggs obtained from groups in pre-molting, post-molting and end of cycle periods were examined for several quality performance traits such as egg weight, specific gravity, shape index, shell strength, shell thickness, eggshell weight, haugh unit, albumen index, yolk index and yolk color. Results indicated that non-feed removal molting programme based particularly on grain barley had positive effect on egg quality traits in laying hens. Notably, yolk color and haugh unit, which are considered as the most important quality parameters from the consumer point of view, were relatively improved in barley molted group.

Induced molting by feed withdrawal has been a common practice in the commercial layer industry and usually involves the removal of feed for a period of up to 14 d. However, this is a practice that is believed to adversely influence the welfare of the hens and there is a need to examine behavioral responses to alternative molt regimens. The behavioral patterns of hens on 90% alfalfa:10% layer ration, 80% alfalfa:20% layer ration, and 70% alfalfa:30% layer ration molt diets were compared with feed withdrawal (FW) hens, and fully fed (FF) hens. The White Leghorn laying hens were approximately 54 wk old and were placed in 3 identical climate-controlled rooms. The hens were individually housed in 2-tier wire battery cages and provided treatment rations and water ad libitum. Nonnutritive pecking, walking, drinking, feeder activity, preening, aggression, and head movement were quantified during two 10-min periods each day for 6 hens from each treatment. Over the 9-d treatment period, hens in the FW, 70% alfalfa:30% layer ration, and 80% alfalfa:20% layer ration groups spent significantly more time walking than hens in the 90% alfalfa:10% layer ration group. The FF and 70% alfalfa:30% layer ration hens spent half as much time preening, whereas the FW hens displayed nearly twice as much nonnutritive pecking when compared with other treatments. Most differences in head movements occurred at the beginning of the molt period, whereas during the last half of molt, alfalfa-fed hens exhibited feeder activity similar to FF hens, and all were significantly higher than that of FW hens. After some initial adjustment by the hens, consumption of alfalfa molt diets appeared to reduce nonnutritive pecking behavior, which is characteristically associated with FW hens.

Moulting is a natural physiological process where the reproductive system of birds undergoes complete remodeling in preparation for the next laying cycle. In domestic chickens, moulting is artificially induced by feed withdrawal to recycle the old laying flock for best profit margins. This has received severe criticism from animal welfare organizations, forcing several countries to stop this practice. Several alternative methods to feed withdrawal methods were developed but were found to produce inconsistent results. Understanding the actual mechanism of moulting would help in designing a new animal welfare friendly method. The present investigation attempted to study the molecular mechanism of moulting in White Leghorn hens. Eighty-four layers (75 weeks) were divided into two groups. The birds in the first group were subjected to moulting by feed withdrawal (FW) while the other group received high dietary Zn (ZnF) treatment for 10 days. Six birds from each group were sacrificed on 0, 1-4, 6 and 10 days of moulting and mRNA expression of caspases-1, -2 and iNOS, along with the apoptotic ladder pattern and nitric oxide (NO) in the ovary and oviduct, was investigated. The mRNA expression of iNOS was upregulated with a corresponding increase in NO levels. Caspases-1 and -2 were differentially upregulated in the ovary and oviduct of moulted birds. A constant decline in serum estradiol and progesterone levels was also observed. It can be concluded that the pattern of reproductive regression during moulting by the two methods is different, as the expression of genes studied in the present investigation is different.

Induced molting of laying hens is a practice used by commercial egg producers to increase the productive lifetime of their flock. However, the conventional method of inducing molt, which involves removal of feed, water, or both as well as a reduction in photoperiod to less than a natural day has drawn criticism due to animal welfare and food safety concerns. The objective of this study was to explore the efficacy of diets containing high levels of guar meal (GM) in inducing molt and reducing susceptibility to Salmonella Enteritidis colonization in late-phase laying hens. Late-phase (68 wk old) Lohmann laying hens were either full-fed standard laying hen diets (nonmolted control), induced to molt by feed withdrawal, or full-fed standard laying hen diets containing 20% GM with or without 250 units/kg of mannanase Hemicell supplementation. On the fourth day of treatment, all hens were orally challenged with SE (1.65 x 10(7) cfu). Hens were killed and evaluated for Salmonella colonization and differences in organ weights 5 d postinoculation. Salmonella Enteritidis present in crop, liver, ovary, and cecal contents were significantly reduced by feeding GM with enzyme supplementation compared with feed withdrawal hens. No significant differences were observed in reproductive tract weights of molted groups, although a difference in liver weight was detected. Results indicate that feeding diets containing 20% GM are as effective as complete feed withdrawal with respect to inducing molt with the added benefit of improved resistance to Salmonella Enteritidis colonization and translocation.

The reproductive remodeling during molting appears to be a complex physiological mechanism regulated by multiple host factors. Lipopolysaccharide (LPS) induced TNF-alpha factor (LITAF) is one of the transcription factors controlling the expression of TNF-alpha and other cytokines. In the present investigation, we studied the involvement of LITAF in the regression of reproductive tissues of molting birds. Semi-quantitative RT-PCR analysis revealed that LITAF mRNA was generally expressed in both ovary and oviduct. In the molting birds, i.e. those subjected to feed withdrawal (FW) or fed high levels of zinc (ZnF) birds, the LITAF expression was upregulated significantly in the ovary after 4 days of molting (DOM). However, LITAF mRNA levels were three-fold higher in ZnF birds, which might be responsible for a greater degree of follicular atresia. In the oviduct of FW birds, peak LITAF expression was noticed on 4DOM and the levels remained significantly higher until the end of the experiment. In ZnF birds, LITAF expression reached its peak on 1DOM and subsequently downregulated to basal levels on 2DOM. This indicated that constantly higher LITAF expression might be required for complete regression of the oviduct during molting. In conclusion, LITAF might be one of the major transcription factors controlling reproductive regression in chicken, as the expression levels were associated with the regression pattern.

Immunological responses of molting hens either infected or not infected with Salmonella enterica serovar Enteritidis were compared in 2 trials with Single Comb White Leghorn hens >50 wk old. The hens were placed into 6 treatment groups with 12 hens per group: nonmolted Salmonella Enteritidis positive (FF+), non-molted Salmonella Enteritidis negative (FF-), feed withdrawal Salmonella Enteritidis positive (FW+), FW Salmonella Enteritidis negative (FW-), alfalfa Salmonella Enteritidis positive (ALC+), and ALC Salmonella Enteritidis negative (ALC-). Each hen in the Salmonella Enteritidis-positive groups was challenged on d 4 of the study with 1 mL of 10(6)-cfu Salmonella Enteritidis, and diets were administered for 12 d. Blood samples were collected on d 2, 5, 9, and 12, and blood smears were enumerated for heterophil to lymphocyte (H:L) ratios. Serum samples were also analyzed for alpha(1)-acid glycoprotein (AGP) levels and antibody level. On d 12, hens were euthanized and bile samples from the gall bladder and sections of the ileum and the ceca were collected, and an ELISA was used to determine the intestinal, serum, and bile antibody responses. The FW+ hens produced more (P

The objective of this study was to examine an alfalfa crumble diet as an alternative molt diet and compare the physiological response of hens to the responses of feed-deprived molted hens. Hens >50 wk old were placed into 6 treatment groups (12 hens per group in trial 1 and 10 hens per group in trial 2): nonmolted Salmonella enterica serovar Enteritidis positive (FF+), nonmolted Salmonella Enteritidis negative (FF-), feed withdrawal Salmonella Enteritidis positive (FW+), FW Salmonella Enteritidis negative (FW), alfalfa Salmonella Enteritidis positive (ALC+), and ALC Salmonella Enteritidis negative (ALC-). Each hen in the Salmonella Enteritidis-positive groups was challenged on the fourth day of the study with 1 mL of 10(6)-cfu Salmonella Enteritidis. Blood was collected on d 2, 5, 9, and 12 of the trial. Blood plasma was collected and metabolite concentrations were analyzed for glucose, calcium, cholesterol, uric acid, total protein, and triglycerides. The feed intakes of the FF hens were 4- to 6-fold greater (P

The objective of this study was to examine microbial population shifts and short-chain fatty acid (SCFA) responses in the gastrointestinal tract of Salmonella Enteritidis-challenged molted and nonmolted hens fed different dietary regimens. Fifteen Salmonella-free Single Comb Leghorn hens (>50 wk old) were assigned to 3 treatment groups of 5 birds each based on diet in 2 trials: 100% alfalfa crumbles (ALC), full-fed (FF, nonmolted) 100% commercial layer ration, and feed withdrawal (FW). A forced molt was induced by either a 12-d alfalfa diet or FW. In all treatment groups, each hen was challenged by crop gavage orally 4 d after molt induction with a 1-mL inoculum containing 10(6) cfu of Salmonella Enteritidis. Fecal and cecal samples (d 4, 6, 8, 11, and necropsy on d 12) were collected postchallenge. Microbial population shifts were evaluated by PCR-based 16S ribosomal RNA gene amplification and denaturing gradient gel electrophoresis, and SCFA concentrations were measured. Total SCFA in fecal and cecal contents for FW molted hens were generally lower (P < or = 0.05) in the later stages of the molt period when compared to ALC and FF treatment groups. The overall trend of SCFA in cecal and fecal samples exhibited similar patterns. In trials 1 and 2, hens molted with ALC diet generally yielded more similar amplicon band patterns with the FF hens in both fecal and cecal samples by the end of the molting period than with FW hens. The results of these studies suggest that ALC molted hens supported microflora and fermentation activities, which were more comparable to FF hens than FW hens by the end of the molting period.

Four layer genetic stocks consisting of 3 Ottawa control strains (5, 7, and 10) and a commercial laying stock (CCS) were utilized to evaluate potential changes in behavioral profiles due to the effects of genetic selection through 2 production cycles. The Ottawa strains were started as random bred strains from the crosses of several popular commercial layers in 1950, 1959, and 1972, and the commercial strain used herein was from calendar year 1993, and its ancestors were involved in the formation of all of the random bred strains. The behavior study utilized 2 replicates from each strain that contained 4 cages, 6 hens/cage, for a total of 192 hens. Behavioral observations were recorded on 2 consecutive days beginning at 22 wk of age and every 28 d thereafter during the first production cycle, the molt period, and the second production cycle through 90 wk of age and periodic feather and Hansen's test scores recorded. Behavior profiles were similar between the control strains and the CCS, indicating that long-term genetic selection by commercial egg-type breeding firms to enhance production parameters has had no impact on laying strain behavior patterns. Appetitive behaviors were not affected by strain. During the molt, hens had reduced (P < 0.05) feeding and drinking frequencies in comparison with those observed during the first and second cycles. The data indicated that hens pecked inedible objects at a greater (P < 0.0001) frequency during the first cycle and molt than during the second cycle. Fearfulness scores were only influenced by production phase with the molt having the highest (P < 0.01) score of 3.46. Strain or production phase did not influence the frequency of aggressive and submissive acts.

The molting of commercial layers has been under increased scrutiny by animal rights groups, who have said that this practice is highly stressful and one which negatively affects the welfare of the hen due to the initial period of fasting that has been used to stop egg production. In recent years, there has been a recognized need to develop practical alternatives to molting layers other than the use of fasting. Thus, the University of Illinois, University of Nebraska, North Carolina State University, and the University of California have all researched this area. In all of these studies, the methods involved comparing a normal fasting method (i.e., 5 to 13 d), to feeding low-energy and protein diets using ingredients such as wheat middlings, soybean hulls, and corn or diets with graded levels of added salt and without salt (University of Nebraska, University of California). The molt period (28 d) included full-feeding of these diets. In these studies, postmolt production performance for the nonfeed withdrawal techniques was comparable to the fasting method. Several researchers have also evaluated the behavioral repertoire of laying hens, which includes feeding, drinking, comfort, social, reproductive, and anti-predator behaviors. In addition, related behaviors such as aggression, escape-avoidance, and submission have been of particular interest as potential indicators of welfare during molting. In these studies, genetic selection, strain, density, or molt program do not appear to adversely influence the behavioral patterns during the molt. The behavior patterns displayed during a molt program appear consistent with the response to physiological changes that layers experience and do not appear to compromise the welfare status of the hens. Appetitive behaviors were not affected by strain but were affected by production phase and molting. Strain or production phase did not influence the frequency of aggressive and submissive acts. Thus, the use of alternative nonfeed withdrawal molting methods provide comparable laying hen well-being and may enhance the transition from a productive to a resting state.

Two experiments of 4 x 2 x 2 factorial arrangements of 4 dietary energy levels, 2 molting methods (feed withdrawal and no salt diet), and 2 strains (Bovans White and Dekalb White) were conducted to determine the effect of dietary energy and molting method on long-term postmolt performance of 2 strains of commercial Leghorns. In experiments 1 and 2, Bovans White hens (n = 576) and Dekalb White hens (n = 576) were randomly divided into 16 treatments (6 replicates of 12 birds per treatment). Experiment 1 lasted from 86 to 96 wk of age, and experiment 2 lasted from 100 to 110 wk of age. Bovans White hens had significantly higher egg production than Dekalb White hens, whereas Bovans White hens had significantly lower egg weight, percentage of eggshell, and egg specific gravity than Dekalb White hens. Based on improved feed conversion, dietary energy of 2,846 kcal of ME/kg appeared to be enough for optimal performance during second cycle phase 2. Based on BW of hens, dietary energy level for optimal performance should be less than 2,936 kcal of ME/kg during second cycle phase 3. There can be no fixed ideal dietary energy level for optimal profits for postmolt egg production. Molting method had no effect on egg production and egg mass during the early and middle stages of the postmolt production period. However, hens molted by feed withdrawal had significantly higher egg production and egg mass during the later stage of the postmolt production period compared with hens molted by a no salt diet. There was no significant difference in egg specific gravity due to molting method. Feeding a no salt diet resulted in reasonable long-term postmolt performance and eggshell quality, rather than optimal performance and eggshell quality.

The aim of this study was to evaluate effects of alfalfa and fructooligosaccharides (FOS) on molting performance and bone parameters compared with the conventional feed withdrawal molting procedure. A total of 36 Single Comb White Leghorn hens (84 wk of age) were used for this experiment. The hens were divided into 6 treatment groups with 6 birds per treatment: pre-trial control (PC), full fed (FF), feed withdrawal (FW), 100% alfalfa (A100), A100 + 0.375% FOS (A100L), and A100 + 0.75% FOS (A100H). At the end of the 9-d molt period, hens were euthanized, and tibia and femurs were collected to evaluate bone qualities using dual-energy x-ray absorptiometry (DXA), Instron (Model 1011 Instron Universal Testing Machine, Instron Corp., Canton. MA), and conventional bone assays. Egg production was recorded during the molting period to evaluate first day out of production, and ovary was also collected to measure ovary weight. Alfalfa molting diets had comparable molting parameters, such as percentage of BW loss, ovary weight, and first day out of egg production, to the conventional feed withdrawal molting procedure, and FOS supplementation did not have any detrimental effects on molting performance. Conventional bone assay and DXA results suggest that hens lose a considerable amount of bone minerals during a molting period. The tibia and femur bone strengths of the FF, FW, A100, and A100L hens were significantly lower than the PC hens, whereas hens fed A100H had similar tibia bone breaking strength to that of the PC hens. The bone parameters measured by conventional assays, bone breaking strength measured by Instron, and bone density and mineral content measured by DXA were highly correlated to each other.

Induced molting in egg-laying hens is an important method for maximizing hen egg production and quality as well as hen health in commercial settings; however, there is growing societal concern over its effects on hen well-being. Using individual hens as their own controls, this research examined the behavior of hens subjected to different treatments of induced molting under premolt, molt, and postmolt conditions. Cage pecking increased in fast-induced subjects and aggression increased in fast-induced and nonfast-induced subjects during the molt. Gakel calling and several aspects of its acoustic structure were much higher during the molt condition in fast-induced subjects only. These data suggest that nonfast-induced molting treatments provide an effective method for inducing molting in hens and improving their well-being by minimizing discomfort due to food deprivation. In addition, these data further support that gakel vocalizations in hens may serve as an effective indicator for assessing well-being in a species otherwise behaviorally stoic in expressing stress or discomfort.

It is important to know how different organs 'manage' their stem cells. Both hair and feather follicles show robust regenerative powers that episodically renew the epithelial organ. However, the evolution of feathers (from reptiles to birds) and hairs (from reptiles to mammals) are independent events and their follicular structures result from convergent evolution. Because feathers do not have the anatomical equivalent of a hair follicle bulge, we are interested in determining where their stem cells are localized. By applying long-term label retention, transplantation and DiI tracing to map stem cell activities, here we show that feather follicles contain slow-cycling long-term label-retaining cells (LRCs), transient amplifying cells and differentiating keratinocytes. Each population, located in anatomically distinct regions, undergoes dynamic homeostasis during the feather cycle. In the growing follicle, LRCs are enriched in a 'collar bulge' niche. In the moulting follicle, LRCs shift to populate a papillar ectoderm niche near the dermal papilla. On transplantation, LRCs show multipotentiality. In a three-dimensional view, LRCs are configured as a ring that is horizontally placed in radially symmetric feathers but tilted in bilaterally symmetric feathers. The changing topology of stem cell activities may contribute to the construction of complex feather forms.

This study investigated changes in bone integrity and circulating concentrations of insulin-like growth factor-I (IGF-I) of hens subjected to 2 distinct molting regimens and fed pre- and postmolt diets high in n-3 or n-6 fatty acids. A dual-energy x-ray absorptiometer determined bone mineral density (BMD) of the tibia and humerus of 45 live hens from 62 to 76 wk of age. Densitometric scans were also conducted in excised tibia and humerus at 66, 71, and 76 wk of age. Concentrations of IGF-I were monitored using an homologous RIA at the same ages. The molting treatments consisted of 10 d of fasting + cracked corn for 7 d + pullet developer diet for 10 d or a nonfasting molt (wheat-middlings-based diet for 27 d). Five weeks prior to and after either molt treatment, birds were fed 1 of 2 diets containing dietary n-6/ n-3 fatty acids ratios of 0.6 or 8.0. At the end of the molt (71 wk of age), tibial BMD decreased 30% in fasted and 11% in nonfasted molt regimens, and the fatty acid content of the premolt diet had no effect on the decline in BMD. The BMD of the humerus also decreased during molt with the exception of hens subjected to a nonfasted molt and fed n-3 fatty acid diets in which their BMD values were similar to or greater (at 73 wk of age) than those of controls during the entire experimental period (treatment by bone by age, P < or = 0.0001). Induced molt affected circulating IGF-I concentrations (treatment by age interaction, P < or = 0.0001), and the response was the same regardless of molt regimen (fasting vs. nonfasting) or diet (n-3 vs. n-6 fatty acids). A decrease in IGF-I 54 h postmolt was noted; however, from 13 to 43 d postmolt, all molted birds had elevated IGF-I as compared with controls. In conclusion, a nonfasted molt as compared with fasted molt was less detrimental to bone mineralization; dietary n-6/n-3 fatty acid ratios in the pre- and postmolt diets had little effect on the decline of skeletal integrity during molt, and circulating IGF-I concentrations were affected by molt.

To determine changes in bone mineralization during molt, 66-wk-old White Leghorns were assigned to either a fasted molting regimen using feed removal for 10 d, followed by the ad libitum consumption of cracked corn for 7 d and a pullet developer diet for 10 d or a nonfasted molting regimen lasting 27 d that included the ad libitum consumption of a diet containing 71% wheat middlings and 23% corn. Both molting regimens restricted light to 8 h/d, and water was provided ad libitum. At 28 d postmolt, hens from both molting treatments were returned to a regular egg-laying diet and 16 h/d of photoperiod. Control hens consumed a regular egg laying diet and were kept on 16 h/d of light throughout the study. Using dual-energy x-ray absorptiometry, bone mineral density (BMD) and bone mineral content (BMC) of the left tibia were measured in 7 live hens per treatment immediately prior to, during, and following the molt. Results showed that by 28 d postmolt, BW loss was 22 and 18% in the fasted and nonfasted molting regimens, respectively (P < or = 0.0001). Compared with premolt values, tibial BMD at 28 d postmolt decreased 35 and 18% in the fasted and nonfasted molt groups, respectively (treatment x age interaction, P < or = 0.0001). Similarly, tibial BMC values decreased 39 and 27% in the fasted and nonfasted molt groups, respectively (treatment by age interaction, P < or = 0.01). The tibial BMD and BMC of controls at 28 d postmolt were similar to premolt values. Recovery in tibial BMD and BMC of fasted and nonfasted hens occurred by 126 d postmolt with values similar to controls. These results suggest that a nonfasted molting regimen is less deleterious to tibial BMD and BMC than a fasted molting regimen.

The method most commonly used to induce molting and stimulate multiple egg-laying cycles in laying hens for commercial egg production is to fast the hens. Unfortunately, increased risk of Salmonella enteritidis (SE) infection may result from the use of this method. Methods to stimulate multiple egg-laying cycles without increasing the risk of SE infection are needed. Hens over 50 wk of age were divided into 12 groups of 11 hens each and placed in individual laying cages. One week prior to dietary changes, hens were placed on an 8-h light and 16-h dark photoperiod that continued for the 9-d molt. All hens were challenged orally with 10(6) cfu of SE on the fourth day of the molt. Treatments were nonfed hens with distilled water (NFD), nonfed hens with the experimental chlorate product (ECP, which provided 15 mM chlorate ion) water (NFECP), alfalfa diets with distilled water (ALD), and alfalfa diets with ECP water (ALECP). In the NFD hens, 67% (log10 2.74) of the crops and 94% (log10 5.62) of the ceca were colonized, whereas for the NFECP hens significant reductions to 22% (log10 1.05) of the crops and 61% (log10 2.44) of the ceca were observed. In the ALD hens, 61% (log10 2.52) of the crops and 94% (log10 4.06) of the ceca were colonized. In the ALECP hens, highly significant reductions to 11% (log10 1.26) of the crops and 39% (log10 1.12) of the ceca were observed. When compared with the NFD hens, significant reductions in SE invasion of the ovary, liver, and spleen occurred in all other treatments, except the ovary in the ALD hens. The low alfalfa intake is probably a factor in our lowered protection against SE when compared with previous results. For several parameters, these results suggest that ECP or the combination of ECP and alfalfa may be a useful tool to reduce the risk of SE during an induced molt.

The effect of moult on eggshell mechanical properties, on composition and concentrations of organic matrix components and on eggshell microstructure was investigated. The observed changes were studied to understand the role of organic matrix and eggshell microstructure in eggshell strength. Moult was induced by zinc oxide (20 g zinc/kg diet) in 53 ISA Brown laying hens at 78 weeks of age. No difference was observed for egg or eggshell weights after moult. In contrast, moult improved the shell breaking strength (28.09 vs 33.71 N). After moult, there was a decrease in the average size of calcite crystals composing the eggshell and in their heterogeneity, whereas crystal orientation remained basically the same. After moulting, the total protein concentration in eggshell increased slightly. The comparisons of SDS-PAGE profiles of the organic matrix constituents extracted before and after moulting showed changes in staining intensity of certain bands. After moult, bands associated with main proteins specific to eggshell formation (OC-116 and OC-17) showed higher staining intensity, while the intensity of the egg white proteins (ovotransferrin, ovalbumin and lysozyme) decreased. ELISA confirmed the decrease in ovotransferrin after moult. Its concentration was inversely correlated with breaking strength before moult. These observations suggest that changes in eggshell crystal size could be due to changes in organic matrix composition. These changes may provide a mechanism for the improvement in shell solidity after moulting.

The effect of an induced molt and a second egg laying cycle on White Leghorns hen's skeletal integrity was investigated in a series of 3 experiments. Using dual-energy X-ray absorptiometry, bone mineral density (BMD) and bone mineral content (BMC) of the left tibia and humerus were measured in live hens and excised bones and correlated with invasive bone measurement tests, egg traits, and the incidence of broken bones in carcasses of processed hens. The results of all 3 experiments showed that an induced molt was detrimental to skeletal integrity. For hens that were repeatedly scanned throughout the second cycle of lay, the BMD of the humerus never recovered after the molt. Recovery of tibial BMD to premolt values occurred late in the second cycle of lay when egg production was declining. The in vivo BMD scans conducted between 77 and 117 wk of age correlated with bone breaking force and bone ash weight (r = 0.58 and r = 0.65, respectively; P < 0.0001). The percentage of freshly broken bones per bird at the end of processing at 126 wk of age averaged 34% and ranged from 0 to 61%. The incidence of broken bones was negatively correlated with the excised tibial BMD and BMC at 126 wk of age (r = -0.54 and r = -0.53, respectively; P < 0.05). In conclusion, feed withdrawal for 10 d during an induced molt was detrimental to the skeletal integrity of hens, and as BMD and BMC of excised tibia at 126 wk of age decreased in White Leghorns, the incidence of bone breakage increased.

A study was conducted to evaluate skeletal quality and eggshell parameters of molted hens at the end of the second laying cycle. Sixty Single Comb White Leghorn hens were used for this study. There were 2 controls and 4 molting treatments: full-fed control 1 (82 wk old; FF1), full-fed control 2 (122 wk old; FF2), feed withdrawal (FW), 100% alfalfa (A100), 90% alfalfa/10% layer ration (A90), and 70% alfalfa/30% layer ration (A70). At the end of the second laying cycle (approximately 122 wk of age), hens were euthanized by CO2. Tibia and femur were collected. There were no differences in bone parameters between FF1 and FF2 (P > 0.05) hens. There were no differences in bone parameters among the different molting dietary treatments (P > 0.05). In the eggshell parameters, the FF2 hens exhibited heavier egg weights than the FF1 (P < 0.05), whereas the percentage shell and egg production of the FF1 birds were significantly higher than those of the FF2 birds. Shell weights of the FW and A90 birds were significantly heavier than that of the A100. The correlation analysis showed that overall bone parameters were negatively correlated with eggshell parameters. Bone parameters were highly correlated with each other. Shell weight, percentage shell, and shell thickness were positively correlated with each other, whereas egg weight was negatively correlated with percentage shell. These results suggest that age of hens and molting dietary treatments influence egg parameters, and eggshell formation is closely related to bone metabolism in laying hens.

Molting is a common practice used by the commercial egg industry to rejuvenate flocks for a second or third laying cycle. During this time the hens rest from production, and the reproductive organs are rejuvenated to increase production and quality during the next laying cycle. Although feed withdrawal (FW) is the most popular and effective method of molt induction, it has come under scrutiny due to food safety issues and animal welfare issues. This study involved feeding alfalfa mixed with layer ration at different ratios to hens to determine their ability to induce molt. The treatment ratios were 100% alfalfa (A100), 90% alfalfa and 10% layer ration (A90), and 70% alfalfa and 30% layer ration (A70). In addition, a fully fed (FF) nonmolted control and a FW negative control were used. Alfalfa is an insoluble, high fiber feedstuff with low metabolizable energy. Egg production for A90 and FW treatments ceased completely by d 6, whereas birds fed A100 and A70 ceased egg production by d 8. Ovary and oviduct weight of hens fed all molting diets decreased (P < 0.05) by an average of 1.5 to 2.5% (BW basis) compared with FF control during the 9-d molt induction period. As the percentage of layer ration increased, feed intake also increased and percentage of BW loss decreased during the 9-d molt induction period. Hens molted by FW lost an average of 25.8% BW, whereas A70 hens lost 18.9% BW. Nonmolted hens (FF) and A70 treatment hens had significantly lower (P < 0.05) egg production when compared with all other treatments over the 39-wk postmolt period. FF treatment hens also had lower (P < 0.05) albumen heights when compared with all other treatments. From these results, alfalfa or alfalfa mixed with layer ration appears to be viable alternatives to conventional FW methods for the successful induction of molt and retention of postmolt performance.

Molting is a process by which a hen's reproductive tract is rejuvenated prior to the beginning of a laying cycle. This process is often artificially induced in commercial settings in order to extend the productive life of a flock of hens. The most common method for the induction of molt is feed withdrawal for a period of several days. It has been noted that feed withdrawal, while effective in inducing molt and allowing an adequate reproductive rest period for the hen, may cause deleterious effects on the animal. This has prompted the investigation of alternatives to feed deprivation for the induction of molt in commercial laying hens. This study involved feeding alfalfa to hens to assess its ability to induce molt. Results show that alfalfa meal and alfalfa pelleted diets were equally effective as feed withdrawal in causing ovary weight regression in birds. Molted hens induced by alfalfa diets exhibited postmolt levels of egg production over a twelve week period that were similar to that of hens molted by feed withdrawal. The postmolt eggs laid by hens molted by alfalfa were of comparable quality to eggs from feed deprived hens. Alfalfa, a fibrous feed with low metabolizable energy, may be provided to hens on an ad libitum basis for an effective molt induction that retains comparable egg quality and production.

Feed deprivation is used in the layer industry to induce molting and stimulate multiple egg-laying cycles in laying hens. Unfortunately, the stress involved increases susceptibility to Salmonella enteritidis (SE), the risk of SE-positive eggs, and incidence of SE in internal organs. Leghorn hens over 50 wk of age were divided into 4 treatment groups of 12 hens each in experiment 1 and 3 treatment groups of 12 hens in experiments 2 and 3; hens were placed in individual laying hen cages. Treatment groups were 1) nonmolted (NM) and received feed and distilled water for 9 d, 2) force molted by feed removal for 9 d and received distilled water, 3) force molted by feed removal for 9 d and received 0.5% lactic acid (LA) in distilled water. An additional group (4) in experiment 1 only was force molted by feed removal for 9 d and received 0.5% acetic acid in distilled water. Seven days before feed removal hens were exposed to an 8L:16D photoperiod, which was continued throughout the experiment. Individual hens among all treatments were challenged orally with 10(4) SE on d 4 of feed removal. When compared with the NM treatments, weight losses were significantly higher in the M treatments, regardless of water treatments. When compared with NM treatments, crop pH was significantly higher in the M treatment receiving distilled water. Crop pH was reduced to that of the NM controls by 0.5% acetic acid in the drinking water. No consistent significant changes were observed for volatile fatty acids. The number of hens positive for SE in crop and ceca after culture and the number of SE per crop and per gram of cecal contents were higher in the M treatments, when compared with the NM treatments, but there was no effect of addition of either of the acids to the drinking water. Additional research using different acid treatment regimens may provide a tool for reducing the incidence of SE in eggs and internal organs during and following molting of laying hens.

The uterine (shell gland) epithelium from the domestic chicken was mounted in Ussing chambers, bathed in symmetric avian saline solution on both apical and basolateral aspects and voltage clamped at 0 mV. The epithelium exhibited a basal short circuit current (I(sc)) that was partially inhibited by the epithelial Na(+) channel (ENaC) blockers, amiloride and benzamil (IC(50) values of 0.8 and 0.12 micromol l(-1), respectively). Inhibition of basal Na(+) absorption by 10 micromol l(-1) amiloride was confirmed by measurements of transepithelial Na(+) and Cl(-) fluxes, where inhibition of the apical-to-basolateral and net Na(+) flux occurred, but no significant effects on Cl(-) fluxes were detected. The amiloride-insensitive portion of the basal I(sc) was both Cl(-) and HCO(3)(-) dependent and was inhibited by the Cl(-) channel blocker, diphenyl-2-carboxylate (DPC; 100 micromol l(-1)). Stimulation with 8-(4-chlorophenylthio)-cyclic 3'-5', adenosine monophosphate (8-cpt cAMP) produced a sustained increase in I(sc) that was dependent on both Cl(-) and HCO(3)(-). The magnitude of the amiloride-sensitive I(sc) was approximately twofold greater in birds where shell formation was complete, but oviposition had not yet occurred. In addition, the amiloride-sensitive I(sc) was greater in hens over the age of 55 weeks and in molting birds. The anion-dependent component of the basal I(sc) was reduced in older birds, and electrogenic HCO(3)(-) transport was nearly absent in molting birds. These results demonstrated that electrogenic Na(+) transport in avian shell gland was similar to the mammalian uterine epithelium and increased with age and during molting. Electrogenic Cl(-) and HCO(3)(-) transport were coupled under basal and cAMP stimulated conditions and basal anion transport decreased with age and during molting.

Feed was totally withdrawn from laying hens (n = 30, Hy-Line Brown, 608 d of age, 2.04 +/- 0.07 kg of mean BW) to induce molting. Ten birds were slaughtered on d 0 and 12, and the hepatic and myocardial triacylglycerol (TAG) and phospholipid (PL) fatty acid composition, as well as the tissue malondialdehyde (MDA) and reduced glutathione (GSH) concentrations were determined. The liver TAG and PL contents decreased by 24.3 and 16.1%, respectively, whereas the myocardial TAG content increased by 12%, and the PL decreased by 22%. Liver TAG fraction has been found to selectively retain arachidonic and docosahexanoic acids. Hepatic PL fatty acids were markedly affected by fasting; these changes reflected an altered PL metabolism, primarily degradation. Liver TAG compensated for the absence of dietary fatty acids, because we found practically no qualitative alteration in myocardial TAG. The lipid peroxide status, as measured with MDA content was, accordingly, increased in the liver tissue only. In the myocardial PL fatty acids, preferred conservation of arachidonic acid was shown, and it was hypothesized that energy deprivation of cardiomyocytes strongly improved PL degradation in fasting laying hens and influenced PL homeostasis. Generally the physiological recovery from forced molting associated with fasting is complete; however, the use of total feed withdrawal methods should be reevaluated.

Three experiments were conducted to define the relationships between BW and feed intake in broiler breeder female chickens at the onset of lay and after a forced molt. In experiment 1A the number of normal yellow follicles at the onset of lay and abdominal fatness were linearly related to BW, whereas age at first egg decreased with increasing BW in a curvilinear manner. Dietary energy was shown to be the limiting factor determining the onset of lay in severely feed restricted broiler breeders in experiment 1B. The source of energy (fat or carbohydrate) did not affect the onset of lay or ovarian function. In experiment 2 birds were feed restricted or fed ad libitum to 60 wk of age and force molted. Eight weeks after the initiation of molting, restricted birds were fed to 3 or 4 kg, and the birds fed ad libitum were fed to 4 or 5 kg of BW. After photostimulation the birds were fed ad libitum or 115 g/d. Taken together, the results of these analyses suggest that rearing treatment did not affect ovarian function, postmolt. Postmolt feed restriction was associated with lower follicle numbers compared with the groups fed ad libitum and was significantly lower in the lighter (3 kg) restricted birds compared with the other groups. Follicle numbers were lower after molting than at the onset of lay, and the effects of feed restriction were commensurately smaller.

1. The effect of age on ovarian function was studied in 245-, 350-, 500-, 700- and 800-d-old Lohmann hens. The effect of three different methods for moult induction on ovarian function and corticosterone concentration was studied in 500-d-old hens. 2. No significant reductions in ovarian weight or in number of follicles before the age of 700 d were found. The ability to produce progesterone and oestradiol-17beta was unchanged up to the age of 700 d and the circadian secretion of these two steroids was identical in young (225 d) and old hens (600 d). 3. The effects of induced moulting by feed withdrawal (FW) and a high Zn (HZn) diet on body weight and ovarian function were very similar; those of a moderate Zn with low Ca (MZn/LCa) diet were smaller. 4. The first significant effect of moulting was a decrease in oestradiol-17beta plasma concentration (d 2). Plasma progesterone decreased more gradually than oestradiol-17beta, and reached a nadir on d 6 in FW- and HZn-treated hens and on d 9 in MZn/LCa-treated ones. 5. Hens treated with either FW or the MZn/LCa, but not those with the HZn diet, showed a very sharp rise in corticosterone concentration on d 2 of treatment. Thus the MZn/LCa diet was less efficient than the other treatments in induction of ovarian involution, but had a similar effect on stress induction, as indicated by increases in plasma corticosterone.