Search

Parental exposure to insults was initially considered safe if stopped before conception. In the present investigation, paternal or maternal preconception exposure to the neuroteratogen chlorpyrifos was investigated in a well-controlled avian model (Fayoumi) and compared to pre-hatch exposure focusing on molecular alterations. The investigation included the analysis of several neurogenesis, neurotransmission, epigenetic and microRNA genes. A significant decrease in the vesicular acetylcholine transporter (SLC18A3) expression was detected in the female offspring in the three investigated models: paternal (57.7%, p < 0.05), maternal (36%, p < 0.05) and pre-hatch (35.6%, p < 0.05). Paternal exposure to chlorpyrifos also led to a significant increase in brain-derived neurotrophic factor (BDNF) gene expression mainly in the female offspring (27.6%, p < 0.005), while its targeting microRNA, miR-10a, was similarly decreased in both female (50.5%, p < 0.05) and male (56%, p < 0.05) offspring. Doublecortin’s (DCX) targeting microRNA, miR-29a, was decreased in the offspring after maternal preconception exposure to chlorpyrifos (39.8%, p < 0.05). Finally, pre-hatch exposure to chlorpyrifos led to a significant increase in protein kinase C beta (PKCß; 44.1%, p < 0.05), methyl-CpG-binding domain protein 2 (MBD2; 44%, p < 0.01) and 3 (MBD3; 33%, p < 0.05) genes expression in the offspring. Although extensive studies are required to establish a mechanism–phenotype relationship, it should be noted that the current investigation does not include phenotype assessment in the offspring.

Targeted green light photostimulation during the last stage of broiler incubation increases expression of the somatotropic axis. The purpose of this study was to further shorten the in ovo green light photostimulation and determine the critical age for photostimulation in broilers embryos, as a future strategy for broiler incubation. Fertile broilers eggs (n = 420) were divided into 5 treatment groups. The first group was incubated under standard conditions (in the dark) as the negative control group. The second was incubated under intermittent monochromatic green light using light-emitting diode lamps with an intensity of 0.1 W/m2 at shell level from embryonic day (ED) 0 of incubation until hatch, as a positive control. The third, fourth, and fifth groups were incubated under intermittent monochromatic green light from ED 15, 16, and 18 of incubation, respectively, until hatch. All treatment groups showed elevated somatotropic axis expression compared with the negative control, with the group incubated under monochromatic green light from ED 18 until hatch showing results closest to the positive control. This suggests that broiler embryos can be exposed to in ovo green light photostimulation from a late stage of incubation (when transferring the eggs to the hatchery) and exhibit essentially the same outcome as obtained by photostimulation during the entire incubation period.

SUMMARY Physiological stress is known to promote economic losses by causing immunosuppression. Various factors induce stress, including non-optimal management and infection with various pathogens. The concentration of the stress hormone, corticosterone, is indicative of stress in birds. However, its measurement is difficult due to its elevation caused by catching, handling, bleeding, short half-life in the blood, and short secretion period in feces. We describe a novel method to assess corticosterone concentration by sampling feathers. The viruses studied were 2 immunosuppressive viruses, Marek's disease virus and chicken anemia virus, the respiratory viruses, influenza virus H9N2, Newcastle disease virus, and Infectious bronchitis virus. The study contributes novelty as virus-induced stress by feather examination was described sporadically in chickens previously, and feathers were used to detect corticosterone mainly in dead wild and captive birds. Also, the development of extraction method from feathers is novel by using PBS instead of methanol, and by preparing the feather homogenate with steroid displacement reagent. The study findings revealed that virus infections increased the corticosterone concentrations in feather tips to various extents, from 2× to 10× fold, demonstrating that birds endure stress in addition to the clinical and pathological effects induced by the virus infections.

Molecular mechanisms of seasonal changes in testicular morphology, histoarchitecture, and functions in birds have not been fully elucidated. The aim of the study was to examine the alternations in cell proliferation and apoptosis, these processes-related gene expressions, and gap junction protein (GJA1gene, connexin 43 protein; Cx43) expression and localization in gander testes during annual stages. Testes from domestic ganders (n = 28) in the first reproduction season were obtained at five stages, i.e., prebreeding (PrB), peak of reproduction (PR), postbreeding (PoB), nonbreeding (NB), and onset of reproduction (OR). Males were kept under controlled breeding conditions. Testicular weight, morphometry, and histology (HE staining) were evaluated, and the following parameters were tested: (1) number of proliferating (PCNA-positive) and apoptotic (TUNEL-positive) cells; (2) mRNA and protein abundances of PCNA, caspase-3, and Cx43 by qRT-PCR and Western blot, respectively; (3) activity of caspase-3 by fluorometric method; and (4) localization of Cx43 by immunofluoresence. Testicular weight was found to decrease by 4-fold at the NB stage with massive depletion of germ cells concomitantly with a reduction of seminiferous tubule (ST) diameter and ST lumen compared to the PR and OR stages. The number of proliferating germ cells was higher at PrB and PR than at the PoB stage, whereas the number of apoptotic cells was higher at PoB and NB compared to PrB and OR. Thus, proliferation-to-apoptosis ratios were lower in PoB and NB than other stages. Moreover, mRNA expression of PCNA exhibited down regulation in these stages compared to PrB and PR. Stage-dependent changes in the Cx43 mRNA level and in the localization pattern in the ST germinal epithelium were observed. Lower abundances of GJA1 transcript during NB and OR than at PoB the stage and irregular distribution of Cx43 protein located near the lumen of ST primarily at PrB and NB compared to the remaining stages were noted. These results suggested that during gander testis development, function, regression, and recrudescence, the interaction between processes of cell proliferation, apoptosis, and changes in the localization of Cx43 protein in the germinal epithelium occurred. The balance between these processes may determine the final functional activity or inactive stage of the testes connected with weight and histoarchitecture changes.

Artificial targeted illumination has a pivotal role in reproductive processes of poultry. The light-absorption mechanism in birds consists of 2 main components: the eye (retinal photoreceptors) and extraretinal photoreceptors located in the brain. Previous studies conducted on hens have shown that photostimulation of brain extraretinal photoreceptors elevates reproductive activity, whereas retinal photostimulation suppresses it. We tested the effect of targeted differential photostimulation (TDP) on reproductive activities of broiler breeder males. Fifty broiler breeder roosters (Ross), 21 wk of age, were divided into 5 environmentally controlled light-treatment rooms (n = 10) equipped with individual cages. Rooms 1 and 2 had 2 parallel lighting systems consisting of red light (630 nm) and green light (514 nm), and rooms 3 and 4 had parallel red and blue (456 nm) lighting systems. Room 5, illuminated with white light, served as the control. Birds of all groups were kept under short day (6L:18D) for 2 wk with both lighting systems in each treatment room turned on. At 23 wk of age, birds were photostimulated by gradually increasing one of the lighting systems to 14 h of light in each room, while the other lighting system was left on short day (6L:18D). Weekly semen samples were collected until 65 wk of age and analyzed for volume, motility, concentration and vitality. Monthly blood samples were drawn for plasma hormone assays. At 65 wk of age, roosters were euthanized and hypothalamus, pituitary gland, retina and testes samples were taken for mRNA expression analysis. TDP using long-day red light and short-day green light significantly increased reproductive performance, manifested by higher semen volume, motility and concentration, and testis weight; furthermore, this group had higher plasma testosterone levels, higher GnRH mRNA expression in the hypothalamus, lower levels of aromatase in the testes, and lower mRNA expression of hypothalamic serotonin transporter, and of pituitary prolactin and its receptors in the testes. This is the first study showing a positive effect of TDP on reproduction of broiler breeder roosters.

Targeted in ovo green light (GL) photostimulation during the last days of broiler egg incubation increases embryonic expression of the somatotropic axis, similar to in ovo green light photostimulation from embryonic day (ED) 0 to the end of incubation. The aim of this study was to examine the effect of selected in ovo GL photostimulation periods on post-hatch broiler growth. Four hundred twenty fertile broiler eggs were divided into 7 treatment groups: the first incubated in the dark (standard conditions) as a negative control; the second incubated under monochromatic GL from ED0-ED20 (positive control); the third group incubated under monochromatic GL light from ED15-ED20; the fourth, fifth and sixth groups were incubated under monochromatic GL on ED16, ED17, and ED18, respectively; and the seventh group was incubated under monochromatic GL from ED18-ED20. All illumination was provided intermittently using LED lamps. After hatch, all chicks were transferred to a controlled room under standard rearing conditions. The group incubated under green light from ED18 until hatch showed similar results to the positive control group in body weights, as well as breast muscle weights (as % of body weights), and an elevation in the somatotropic axis activity during the experiment. We suggest that broiler embryos can be exposed to in ovo GL photostimulation from ED18 until hatch (hatching period), and still exhibit the same performance as obtained by photostimulation from d 0 of incubation.

In the mid-1960s egg production, fertility, and hatchability of broiler breeder females dropped precipitously. Due to disrupted follicle hierarchies and development of the erratic oviposition and defective eggs (EODES) syndrome. EODES was controlled by restricting feed. In the 1990s, another set of problems arose at entry of broiler breeders into lay and characterized by high mortality followed by lower peak lay and reduction in egg and chick production. These problems are induced by even slight over-feeding, and hence we termed it the “Over Feeding Complex” (OFC). We have speculated that OFC is a quasi-EODES condition, induced by the intense selection for increased breast proportion. To test this, we compared, under fast (FF) and slow (SF) release from feed restriction, body composition and reproductive performance of a broiler breeder from year 1980 (B1980) and kept without selection for performance traits since then, to a line hatched in 2000 (B2000). During the first 16 d of lay, feeding treatment had little effect on egg mass or Laying % for the B1980 birds, while for the B2000 birds, SF treatment resulted in significantly greater egg mass and Laying % compared to FF, showing that the OFC indeed manifested in this experiment. However, contrary to hypothesis, follicle hierarchies were normal for both lines under both feeding treatments. To gain further insight into the OFC syndrome, we here report levels of estradiol, testosterone, and progesterone for these line and treatment groups in the time period leading up to and into lay. A significant line × feeding treatment interaction effect was found for estradiol and testosterone, to a lesser extent for progesterone. For all 3 hormones, for B1980 levels 2 to 3 wk post entry into lay were similar and intermediate under FF and SF, but differed significantly for B2000, being much greater under SF than under FF. Thus, the hormonal effects were parallel and may explain the egg mass and Laying % effects of FF and SF in the 2 genetic types.

The mesotocinergic (MTergic) and dopaminergic (DAergic) systems have been documented to play pivotal roles in maternal behaviors in native Thai chickens. In native Thai chickens, plasma prolactin (PRL) concentrations are associated with maternal behaviors, which are also controlled by the DAergic system. However, the role of MT in conjunction with the roles of DA and PRL on the neuroendocrine regulation of the transition from incubating to rearing behavior has never been studied. Therefore, the aim of this study was to investigate the association of MT, DA, and PRL during the transition from incubating to rearing behavior in native Thai hens. Using an immunohistochemistry technique, the numbers of MT-immunoreactive (-ir) and tyrosine hydroxylase-ir (TH-ir, a DA marker) neurons were compared between incubating hens (INC; n = 6) and hens for which the incubated eggs were replaced with 3 newly hatched chicks for 3 days after 6, 10, and 14 days of incubation (REC; n = 6). Plasma PRL concentrations were determined by enzyme-linked immunosorbent assay. The results revealed that the numbers of MT-ir neurons within the nucleus supraopticus, pars ventralis (SOv), nucleus preopticus medialis (POM), and nucleus paraventricularis magnocellularis (PVN) increased in the REC hens when compared with those of the INC hens at 3 different time points (at days 9, 13, and 17). On the other hand, the number of TH-ir neurons in the nucleus intramedialis (nI) decreased in the REC13 and REC17 hens when compared with those of the INC hens. However, the number of TH-ir neurons in the nucleus mamillaris lateralis (ML) only decreased in the REC13 hens when compared with the INC13 hens. The decrease in the numbers of TH-ir neurons within the nI and ML is associated with the decrease in the levels of plasma PRL. This study suggests that the presence of either eggs or chicks is the key factor regulating the MTergic system within the SOv, POM, and PVN and the DAergic system within the nI and ML during the transition from incubating to rearing behavior in native Thai chickens. The results further indicate that these two systems play pivotal roles in the transition from incubating to rearing behavior in this equatorial species.

This paper presents the study on determination of gander-goose mating activity in relation to age and the position of gander in social hierarchic rank. Moreover, levels of fertility (F) and testosterone (T) in different gander age groups were analyzed. The study was carried out on one-year-old geese (N = 64) and one- or two-year-old ganders (N = 16; 1♂: 4♀). Observations of mating activity (MA) were conducted during the 3-h period of the daylight (10 h), 2-3 times per week (102 h/ group/34 days). The recording was done with a digital recorder connected to cameras. Agonistic behavioral interactions between ganders were noted to determine a social hierarchic rank. The MA was described by frequency of courtship displays, copulation attempts, successful copulation (SCop), and interaction disrupted (DMI) with agonistic behavior. Fertility was evaluated after the artificial incubation (1×/week) in a commercial hatchery. The frequency of SCop, DMI, and T concentration were higher (P

Previous studies demonstrated that in ovo photostimulation with monochromatic green light increases body weight and accelerates muscle development in broilers. The mechanism in which in ovo photostimulation accelerates growth and muscle development is not clearly understood. The objective of the current study was to define development of the somatotropic axis in the broiler embryo associated with in ovo green light photostimulation. Two-hundred-forty fertile broiler eggs were divided into 2 groups. The first group was incubated under intermittent monochromatic green light using light-emitting diode (LED) lamps with an intensity of 0.1 W\m2 at shell level, and the second group was incubated under dark conditions and served as control. In ovo green light photostimulation increased plasma growth hormone (GH) and prolactin (PRL) levels, as well as hypothalamic growth hormone releasing hormone (GHRH), liver growth hormone receptor (GHR), and insulin-like growth factor-1 (IGF-1) mRNA levels. The in ovo photostimulation did not, however, increase embryo's body weight, breast muscle weight, or liver weight. The results of this study suggest that stimulation with monochromatic green light during incubation increases somatotropic axis expression, as well as plasma prolactin levels, during embryonic development.

Light perception in birds is composed of the retina and extraretinal sites, located in the brain. Previous studies indicate that selective photostimulation of the eye decreased reproductive performance, whereas extraretinal photostimulation increases it. Differential photostimulation of the retina and extraretinal sites is based on the retina's sensitivity to green wavelengths and on the red wavelengths' ability to penetrate body tissues. We previously found that short-day exposure to green light within a long-day exposure to red light increases reproductive activity in female turkeys and broiler breeder hens. Furthermore, in a study conducted recently in our laboratory, we found that blue light repressed expression of green light receptor in the retina, which can further enhance reproduction activity in broiler breeders. Here, we examined the “brain activate/eye deactivate” hypothesis on gonadal axis activity and reproductive performance in a broiler breeder flock. Broiler breeder hens and roosters (ROSS 308) were divided into 5 light-treatment groups (controlled rooms with light-emitting diodes [LED] lamps): warm white (control), long-day (14 h) red (630 nm) and short-day (6 h) green (514 nm) (red-green), long-day green and short-day red (green-red), long-day red and short-day blue (456 nm) (red-blue), and long-day blue and short-day red (blue-red). Birds were reared from 20 to 55 wk of age. Eggs were collected daily. Weekly egg production calculated. All eggs were incubated for fertility and hatchability examination. Blood was drawn monthly for plasma analysis. At 35 wk of age (after peak production) and 55 wk of age (end of the experiment), 10 hens from each treatment group were euthanized, and selected tissues and glands were taken for gene expression trials. Providing long-day red light to extraretinal photoreceptors while maintaining retinal photoreceptors on short day with blue or green light significantly improved reproductive activities, manifested by elevated egg production and gonadal axis activity compared with Controls and primary breeder recommendations. Long-day green light reduced reproductive performances. We suggest that targeted photostimulation enhances reproductive and gonadal axis activities in broiler breeders.

Avian prolactin (PRL) secretion is under stimulatory control by the PRL-releasing factor (PRF), vasoactive intestinal peptide (VIP). The neuroendocrine regulation of the avian reproductive system has been extensively studied in females. However, there are limited data in males. The aim of this study was to elucidate the VIPergic system and its relationship to PRL and testosterone (T) in the male native Thai chicken. The distributions of VIP-immunoreactive (-ir) neurons and fibers were determined by immunohistochemistry. Changes in VIP-ir neurons within the nucleus inferioris hypothalami (IH) and nucleus infundibuli hypothalami (IN) areas were compared across the reproductive stages. Plasma levels of PRL and T were determined by enzyme-linked immunosorbent assay and then compared across the reproductive stages. The results revealed that the highest accumulations of VIP-ir neurons were concentrated only within the IH-IN, and VIP-ir neurons were not detected within other hypothalamic nuclei. Within the IH-IN, VIP-ir neurons were low in premature and aging males and markedly increased in mature males. Changes in VIP-ir neurons within the IH-IN were directly mirrored with changes in PRL and T levels across the reproductive stages. These results suggested that VIP neurons in the IH-IN play a regulatory role in year-round reproductive activity in males. The present study also provides additional evidence that VIP is the PRF in non-seasonal, continuously breeding equatorial species.

Maternal care in birds generally includes incubation, brooding and rearing behavior. The neuroendocrine regulation of such maternal behavior is not fully understood for many bird species. The aim of this study was to elucidate whether changes in the circulating levels of ovarian steroids might be associated with brooding behavior in the female native Thai chicken. Changes in the levels of ovarian steroids of rearing hens were compared with those of non-rearing ones. Plasma estradiol, progesterone, and testosterone levels were determined by enzyme-linked immunosorbent assays. The results revealed differences in circulating levels of ovarian steroids between hens rearing their chicks and those deprived of their young after hatching. The non-rearing hens had higher circulating estradiol, progesteron, and tertosterone levels than those of the rearing hens. These differences were significant during the first two weeks after hatching. The results of the present study demonstrate the inhibitory effect of brooding behavior on ovarian steroids in the native Thai chickens. Indeed, disruption of brooding behavior by removing the chicks from the hens increased circulating levels of ovarian steroids and reinitiated reproductive and egg laying activities.

Reproductive failure associated with aging is a well-known phenomenon. However, the mechanism by which this failure occurs in broiler breeder roosters is still unclear. A previous study conducted in our laboratory, comparing young and aging broiler breeder roosters, demonstrated an elevation in hypothalamic vasoactive intestinal peptide (VIP) and pituitary prolactin (PRL) gene expression accompanied by a deterioration of gonadal axis function. This resulted in a decrease in semen-quality variables as roosters aged. The objective of this study was to examine the involvement of the serotonergic axis in the age-associated reproductive failure in broiler breeder roosters. Cobb roosters aged 64 wk were divided into 3 groups (n = 20 each): parachlorophenylalanine (PCPA) administration, active immunization against chicken VIP, and controls. At 69 wk of age, each group was divided into 2 equal subgroups: 1 received ovine PRL and the other served as controls. Weekly semen volume, concentration and motility, and plasma testosterone, estradiol, and PRL concentrations were examined. At the end of the experiment, roosters were euthanized, testes were weighed, and hypothalamus and pituitary were removed to assay the expression of genes encoding hypothalamic GnRH-I, pituitary FSH, pituitary LH, hypothalamic VIP, and pituitary PRL. Both PCPA administration and active immunization against chicken VIP significantly increased testis weight, semen volume, sperm concentration, ejaculation grade, plasma testosterone level, and GnRH-I, FSH and LH gene expression compared with controls ( P ≤ 0.05). In addition, a decrease in plasma estradiol and PRL concentrations and VIP and PRL gene expression was observed in PCPA- and VIP-immunized birds compared with controls ( P ≤ 0.05). Administration of PRL in all groups decreased gonadal axis function and semen-quality variables ( P ≤ 0.05). Collectively, these results suggest that the increasing expression levels of the serotonergic axis in aging broiler breeder roosters inhibit proper gonadal function and reproductive performance. This article establishes for the first time the inhibitory role of serotonin on reproduction in aging roosters.

The dopaminergic (DAergic) system plays a pivotal role in incubation behavior via the regulation of prolactin (PRL) secretion in birds, however the role of the DA/PRL system in rearing behavior is poorly understood. The objective of this study was to investigate the relationship between the DA/PRL system and rearing behavior in a gallinaceous bird, the native Thai chicken. Incubating native Thai hens were divided into two groups. In the first group, hens were allowed to care for their chicks (rearing hens; R). In the second group, hens were deprived of their chicks immediately after hatching (non-rearing hens; NR). In both groups, blood samples and brain sections were collected at different time points after the chicks hatched (days 4, 7, 10, 14, 17, 21, 24, and 28; 6 hens/time point/group). In this study, tyrosine hydroxylase (TH) was used as a marker for DAergic neurons. The numbers of TH-immunoreactive (-ir) neurons in the nucleus intramedialis (nI) and in the nucleus mamillaris lateralis (ML), which regulate the vasoactive intestinal peptide (VIP)/PRL system, were determined in R and NR hens utilizing immunohistochemical techniques. Plasma PRL levels were determined by enzyme-linked immunosorbent assays. The results revealed that both the number of TH-ir neurons in the nI and the plasma PRL levels were significantly higher in the R hens compared with the NR hens during the first 14 days of chick rearing (P

In breeding geese, natural mating applies. The objective of this study was to determine seasonal- and age-related changes in the sexual activity of ganders. Moreover, T, T4 and T3 plasma levels were evaluated. The study was conducted on 2/3-year-old males (n=15) and 1- or 2/3-year-old females (1♂:4♀). Sexual activity of ganders was characterized through the frequency of: courtship, attempts at and successful copulations and total mating activity (MA). Reproductive results manifested by egg production and fertility were recorded. Laying percentage (January-June) was 37.1 and 28.6% for 1-, and 2/3-year-old geese, respectively. MA was noted before the sexual maturity of females and the T level peak. MA was highest at the onset and during peak production (March) for ganders kept with 1-, and 2/3-year-old geese, respectively. From April to May a decrease in fertility with a reduction of the frequency of copulations was observed. At this time low levels of T were noted. The effect of goose age on the MA was shown, with higher frequency of copulations for ganders kept with 1-year-old geese. We suggest that in the successful seasonal mating in geese, social factors such as the presence of females and female age play an important role. The reduction in fertility during the spring period may be associated with decreases in the efficiency of successive mating sequences. The higher frequency of copulations without affecting fertility, for ganders kept with young geese, may be because of differences in sperm transfer or storage/transport efficiency.

The only light source for chickens in environmentally controlled houses is an artificial one. Thus, source, spectra, intensity and regimen of light supplementation became major factors in modern meat type bird management. Light spectra affect growth in meat type birds both in ovo and post hatch. Broilers photostimulated in ovo with green light gained significantly more weight than birds incubated under dark conditions. Furthermore, we defined the cellular and molecular events associated with the effect of in ovo green photostimulation on muscle growth. We found that in ovo photostimulation have a stimulatory effect on the proliferation and differentiation of satellite cells and a promoting effect on the uniformity of the muscle fibers in the early post-hatch period. How does in ovo photostimulation affect intracellular events, such as proliferation and differentiation of muscle cells, leading to post-hatch muscle growth? It is possible that the monochromatic green light penetrates the eggshell and has a direct effect on the embryo's muscle. We were unable to detect any proliferative effect of monochromatic green light on cultured myoblasts derived from standard (un-illuminated) E17 embryos and 3-day-old chicks. A more likely explanation is that green light indirectly affects myoblast proliferation by activating the endocrine system; the latter receives photic cues from the retinal or extra-retinal photoreceptors. We gathered some evidence to support these findings; we have shown a higher expression of growth hormone (GH) receptor mRNA in satellite cells derived from green light illuminated chicks. In addition, plasma GH levels and IGF-I levels in muscle tissue, were higher in the green group relative to the dark one in early post-hatch. Another possible explanation for this phenomenon could be that growth factor secretion is activated in response to green light photostimulation. Both retinal and extra-retinal photoreceptors are active during embryogenesis and can be first detected at E14. Combinations of in ovo and post-hatch green light photostimulation to broilers and turkeys did not cause synergetic effect on growth. In a recent study, we found that in ovo green light photostimulation suppresses the green and red opsin receptors gene expression in the last three days before hatching, while red light enhances their expression. Furthermore, we found that the down-regulation of the green and red opsins in response to incubation under monochromatic green lighting lasted up to 9days post hatch, suggesting a possible epigenetic effect.

Fertility of domestic roosters decreases at ∼50 wk of age. In a previous study on aging white leghorn roosters, low fertility was accompanied by low levels of both hypothalamic vasoactive intestinal peptide (VIP) and pituitary prolactin (PRL) mRNA expression; however, their role in aging broiler breeder rooster reproduction is still unclear. In this study we compared reproductive activities of young (35-wk-old) and aging (73-wk-old) broiler breeder roosters. Weekly semen volume; concentration and ejaculation grade; and concentrations of plasma testosterone, estradiol, and PRL were examined. Every other week, 10 roosters from each group were euthanized, their testes weighed, and hypothalamus and pituitary removed to determine mRNA expression of hypothalamic GnRH-I, pituitary FSH, pituitary LH, hypothalamic VIP, and pituitary PRL. Aging roosters had significantly lower testis weight and semen volume, sperm concentration, ejaculation grade and plasma testosterone and low hypothalamic GnRH-I, pituitary FSH, and pituitary LH mRNA expression than young roosters (P ≤ 0.05). Aging roosters had higher concentrations of plasma estradiol and PRL and higher hypothalamic VIP and pituitary PRL mRNA expression than young roosters (P ≤ 0.05). We suggest that PRL, which is known to inhibit the gonadal axis, and its releasing factor, VIP, play an important role in the reproductive failure associated with age in broiler breeder roosters.

Decreasing fertility in aging domestic roosters is a well-known phenomenon. Aging is manifested by a decrease in plasma testosterone level, testis function, and spermatogenesis, resulting in a low level of fertility. The roles of vasoactive intestinal peptide (VIP) and testicular inhibin in this aging process are not clear. The effects of active immunization against VIP, inhibin, or the combination of both hormones on the reproduction of aging White Leghorn (WL) roosters were assayed. In experiment 1a, 60 White Leghorn roosters (67 wk of age) were divided into 4 groups (n = 15/group). The first group was actively immunized against VIP, the second against inhibin, the third against VIP and inhibin, and the fourth served as a control. Active immunization against VIP decreased semen quality parameters, plasma steroid levels, and gene expression of gonadotropin-releasing hormone-I (GnRH-I), follicle-stimulating hormone (FSH), luteinizing hormone (LH), LH receptor, VIP, and prolactin (Prl). Immunization against inhibin increased some of the semen quality parameters and FSH mRNA gene expression but decreased inhibin gene expression. In experiment 1b, at 94 wk of age, we took the actively immunized against VIP group and the control group and divided them into 2 subgroups (n = 7 or 8): the first group was injected with 1 mg of ovine Prl (oPrl) daily for 7 d, and the second group served as a control. Administration of oPrl to previously VIP-immunized birds significantly elevated semen quality parameters. We suggest that VIP, Prl, and inhibin have an important effect on the reproductive axis in aging roosters. Active immunization against VIP-depressed reproductive activity and Prl administration restored their reproduction, indicating that both VIP and Prl are essential for reproduction in aging roosters. Immunization against inhibin improved FSH mRNA gene expression, suggesting a negative role of inhibin on FSH secretion in aging roosters. Not all semen quality parameters increased significantly after immunization against inhibin, even though FSH mRNA gene expression increased, suggesting interference in testicular function in aging roosters.

Hyperprolactinemia is associated with incubation behavior and ovarian regression in birds. To investigate the association of prolactin (PRL), vasoactive intestinal peptide (VIP), and dopamine (DA) with the neuroendocrine regulation of incubation behavior, changes in the number of visible VIP-immunoreactive (VIP-ir) neurons in the nucleus inferioris hypothalami (IH) and nucleus infundibuli hypothalami (IN) and tyrosine hydroxylase-immunoreactive (TH-ir) neurons in the nucleus intramedialis (nI) and nucleus mamillaris lateralis (ML) of incubating native Thai hens were compared with those of nest-deprived hens. TH was used as a marker for dopaminergic (DAergic) neurons. Blood samples were collected to determine PRL levels. The localization and the number of visible VIP-ir and TH-ir neurons were determined by immunohistochemistry. Disruption of incubation behavior was accompanied by a precipitous decline in plasma PRL levels. The number of visible VIP-ir neurons in the IH-IN and TH-ir neurons in the nI and ML were high during incubation and decreased when hens were deprived of their nests. This study indicated an association between VIP neurons in the IH-IN and DA neurons in the nI and ML with the degree of hyperprolactinemia, suggesting that the expression of incubation behavior in birds might be, in part, regulated by the DAergic input from the nI and ML to VIP neurons in the IH-IN and subsequent PRL release.

Fertility of bull spermatozoa cryopreserved in large volume by directional freezing technique, thawed, repackaged in straws and refrozen over liquid nitrogen vapor (double freezing, DF) was compared to conventional single freezing in straws (CF). Semen was collected from 6 bulls, 4 of which were selected for the field trial. Each semen collection was split into two parts, one frozen by CF and the other by DF. In vitro semen evaluations included motility (fresh, upon thawing and after 3h incubation at 37 degrees C), viability and acrosome integrity. A total of 3610 cows and heifers were randomly inseminated by either CF or DF at about equal numbers. In vitro sperm analysis indicated no difference between CF and directional freezing in large volume and both were superior to DF (P

The aim of cryopreservation is to maintain cellular integrity, thereby enabling resumption of proper biological functioning after thawing. Here we propose OptiPrep (60% iodixanol in water) as a protectant during sperm cryopreservation using pooled bull semen as the model. We evaluated OptiPrep concentration effect and its relation to cryopreservation by comparing frozen-thawed and chilled samples. Semen, extended in Andromed with 0 (control), 1.25%, 2.5%, and 5% OptiPrep, was compared after either chilling or freezing in large volume by directional freezing. Sample evaluation included sperm motility upon thawing and after 3h incubation at 37 degrees C for frozen-thawed samples and after 3h and 6h of chilling for chilled samples; viability, acrosomal integrity, and hypoosmotic swelling were also tested for frozen-thawed and chilled samples. Chilled samples with 5% OptiPrep showed inferior viability (P=0.047) and 3h motility (P=0.017) relative to that for chilled samples with 2.5% OptiPrep and inferior viability (P=0.042), acrosomal integrity (P=0.045), and 0h motility (P=0.024) relative to that for chilled samples with 1.25% OptiPrep. The 1.25%, 2.5%, and control samples did not differ. In frozen-thawed samples, 2.5% OptiPrep was superior to all other concentrations for 3h motility (control, P=0.007; 5% OptiPrep, P=0.005; 1.25% OptiPrep, P=0.004) and to 1.25% OptiPrep for acrosomal integrity (P=0.001). In a search for a protection mechanism, we measured glass transition temperature (T(g)) of Andromed and of Andromed with 1.25%, 2.5%, and 5% OptiPrep. Andromed (-58.78 degrees C) and 1.25% OptiPrep (-58.75 degrees C) groups had lower mean T(g) than that of the 2.5% (-57.67 degrees C) and the 5% (-57.10 degrees C) groups. Directional cryomicroscopy revealed that the presence of iodixanol alters ice crystal formation into an intricate net of dendrites. Thus, iodixanol appears to possess cryoprotective properties by helping spermatozoa maintain motility and membrane integrity, possibly through altering ice crystals formation into a more hospitable environment and increasing the glass transition temperature.

Native Thai chicken, an equatorial species breeds throughout the year, whereas turkeys are seasonal temperate zone breeder whose reproductive cycle is terminated by the onset of photorefractoriness. This study investigated VIPergic activity throughout a reproductive cycle in both species, hypothesizing that the differential expression of vasoactive intestinal peptide (VIP) would provide an insight into the differing reproductive strategies of the two species. Distribution of VIP neurons in the native Thai chicken and a comparison of VIPergic activity in the nucleus inferioris hypothalami (IH) and nucleus infundibuli hypothalami (IN) were investigated. VIP immunoreactivity was found throughout the native Thai chicken brain, predominantly located within the IH–IN. The pattern of VIP distribution in the native Thai chicken supports the findings reported in temperate zone species. Unlike the turkey, where there is a dissociation between VIPergic activity and prolactin levels during photorefractoriness, in the native Thai chicken, which do not express photorefractoriness, changes in VIP immunoreactive (VIP-ir) neurons within the IH–IN were directly correlated with prolactin throughout the reproductive cycle. VIPergic activity reached its lowest level after hatching of the chicks in the native Thai chicken, while in the turkey VIPergic activity was lowest only after exposure to a short day photoperiod and the acquisition of photosensitivity. This suggests that VIP neurons in the IH–IN may play a pivotal role in regulating the reproductive cycle and its differential expression following hatching of the young may, in part, account for the difference in reproductive mode between equatorial, continually breeding, non-photoperiodic birds and seasonally breeding, photoperiodic birds.

In addition to well recognized effects on the zebra finch song system, we previously showed that post-hatch oral estrogen and xenoestrogen exposure disrupts reproduction by increasing eggshell breakage in females and decreasing fertility in males. Here we show that post-hatch exposure to estradiol benzoate (by oral gavage on days 5 to 11) at a 100 nmol EB per g body mass dose (EB100) also reduces adult oviduct mass. Further, EB100 and doses two orders of magnitude lower (EB10, EB1) reduce egg mass and length. Similar to the induction of song-control nuclei in females, dosing with EB10 and EB100 increased and masculinized another highly differentiated behavior: nest-building. Zebra finches orally exposed as chicks were observed during reproductive trials in communal breeding cages for 4 or 6weeks duration. EB100 males and females and EB10 males showed increased nest-building behaviors. Further, EB10 and EB100 birds had larger nests than canola oil-treated controls, and EB100 birds had faster rates of nest-building than controls, while EB1 birds had significantly slower rates of nest-building than controls. Additionally, EB100 males and females also showed an increased preference for a coarser male-typical nest-building material (jute) over a finer, female-typical material (wool), suggesting a masculinization of nest-building behavior at the higher doses. The change in zebra finch nest-building behavior induced by early EB exposure suggests that nest size and quality, in addition to egg mass and length, may provide new endpoints for assessing avian exposure to xeno- and phyto-estrogens in wild birds.

Reproductive failure associated with heat stress is a well-known phenomenon. The mechanism involved in this failure is not clearly understood. In order to test a possible direct effect of heat stress on ovarian function, 36 White Leghorn laying hens were housed in individual cages in 2 temperature- and light-controlled rooms (n = 18). At 31 wk of age, one group was exposed daily for 12 h to high temperature (42 +/- 3 degrees C), and the second group was maintained under thermoneutral conditions (24 to 26 degrees C) and served as control. Body temperature, feed intake, egg production, and egg weight were recorded daily; heparinized blood samples were drawn every 3 d for plasma hormonal level of luteinizing hormone, follicular stimulating hormone, progesterone, 17beta-estradiol, and testosterone. Six days after exposure half of the birds in each group were killed, and the ovary and oviduct were weighed and preovulatory follicles removed and extracted for mRNA of Cytochrome P 450 aromatase, 17-alpha hydroxylase. The same procedure was repeated 9 d later with the rest of the birds. Short and long heat exposure caused significant hyperthermia and reduction of egg production, egg weight, ovarian weight, and the number of large follicles. In addition, a significant reduction in plasma progesterone and testosterone was detected 2 d after exposing the birds to heat stress, and plasma 17beta-estradiol was significantly reduced 14 d after initiation of heat stress. Short exposure to heat stress caused significant reduction in mRNA expression of cytochrome P450 17-alpha hydroxylase, exposing the birds to long-term heat stress caused significant reduction in expression of mRNA of both steroidogenic enzymes. No significant change was found in plasma luteinizing hormone and follicular stimulating hormone levels during the entire experimental period. We suggest a possible direct effect of heat stress on ovarian function.

Fatty liver hemorrhagic syndrome (FLHS) is a metabolic condition of chicken and other birds caused by diverse nutritional, hormonal, environmental, and metabolic factors. Here we studied the effect of different diet composition on the induction of FLHS in single comb White Leghorn (WL) Hy-line laying hens. Seventy six (76) young WL (26 wks old) laying hens and 69 old hens (84 wks old) of the same breed were each divided into 4 treatment groups and provided 4 different diet treatments. The diet treatments included: control (C), 17.5% CP, 3.5% fat (F); normal protein, high fat (HF), 17.5% CP, 7% F; low protein, normal fat (LP), 13% CP, 3.5% F; and low protein, high fat (LPHF), 13% CP, 6.5% F. The diets containing high fat also had a higher ME of 3,000 kcal/kg of feed while the other 2 diets with normal fat had a regular lower amount of ME (2750 kcal/kg). Hen-day egg production (HDEP), ADFI, BW, egg weight, plasma enzymes indicating liver damage (alkaline phosphatase [ALP], aspartate aminotransferase [AST], gamma-glutamyl transferase [GGT]), liver and abdominal fat weight, liver color score (LCS), liver hemorrhagic score (LHS), liver fat content (LFC), liver histological examination, lipid peroxidation product in the liver, and genes indicating liver inflammation were evaluated. HDEP, ADFI, BW, and egg weight were significantly decreased in the LPHF diet group, while egg weight was also decreased in the LP diet group. In the young hens (LPHF group), ALP was found significantly higher at 30 d of diet treatment and was numerically higher throughout the experiment, while AST was significantly higher at 105 d of treatment. LCS, LHS, and LFC were significantly higher in young hens on the LPHF diet treatment. A liver histological examination shows more lipid vacuolization in the LPHF treatment diet. HF or LP alone had no significant effect on LFC, LHS, or LCS. We suggest that LP in the diet with higher ME from fat can be a possible natural cause for predisposing laying hens to FLHS.

In a natural mating system, the sexual behavior of birds has an important role in fertility success. Non-competitive mating system provides special conditions to study gander-goose interactions. Behavioral and physiological data from a domestic geese breeding flock was used to determine interrelationships between mating activity (MA) parameters, sperm penetration assay (SPA), plasma testosterone (T) concentration, and fertility (F). Moreover, variation in the frequency of gander-goose interactions during the intensive breeding period and the diurnal rhythm (short day - 10L:14D) were evaluated. The 2-/3-year-old ganders (n=15) and 1-/3-year-old geese (1♂:4♀) were observed. On the basis of successful copulation (SCop), groups of ganders with low (33.3%), intermediate (40%), and high (26.7%) frequency were categorized. Frequency of SCop was greater in the morning than in the afternoon. For the entire breeding period, SPA results obtained for the low frequency group were less than for the intermediate group. Fertility success for ganders from both low and intermediate groups was similar. There was a lack of association between MA, plasma T concentration, and SPA results. However, SCop was positively correlated with fertility. It is recommended that SCop be considered as a prognostic parameter for monitoring of ganders' reproductive potential. It is recommended that the sexual behavior of ganders be evaluated before the 1200h of the day. The SCop with an average frequency of 0.4-0.8 times during the day light hours appears to be associated with fertility results that are satisfactory for geese husbandry. Additionally, the SPA may be considered for identification of ganders with poor reproductive potential to facilitate the decision of changes in harem social structure during the first half of the breeding season.

Enhancing meat production by genetic selection for growth has already produced 5-week-old broilers weighing more than 2 kg. As growth performance characteristics continue to improve and the time it takes to achieve market size decreases, the period of embryonic development becomes a greater proportion of the bird's life. Therefore, in parallel to genetic selection, other approaches, such as environmental manipulations in the embryo or in the early days posthatch, are becoming more relevant for increasing muscle growth and meat production.Recently, we have shown that nutritional treatments, i.e., providing feed immediately posthatch, or environmental treatments, such as heat conditioning or monochromatic green-light illumination during the first days posthatch, increase muscle growth and breast muscle weight at marketing day. In all cases, the increase in muscle growth was due to changes at the cellular and molecular levels leading to increased satellite cell proliferation and differentiation.The significant effects on muscle growth resulting from the treatments in the first days posthatch raised the hypothesis that muscle growth could be affected during the embryonic development. In experiments in which eggs were illuminated under monochromatic green light from embryonic day 5 (E5), there was a positive effect on embryo development and posthatch muscle growth. Further studies revealed that this enhanced muscle weight was due to increased satellite cell number and fiber synchronization during early days posthatch. Thermal manipulation at 38.5°C from E16 to E18 for 3 h/day had a delayed effect on satellite-cell proliferation and differentiation, resulting in enhanced hypertrophy of myofibers at market age.

In birds, changes in hypothalamic gonadotropin-releasing hormone-I (GnRH-I) content and release are correlated with reproductive stages. This study examined the distribution and expression level of GnRH-I mRNA in anatomically discrete hypothalamic nuclei throughout the turkey reproductive cycle and following photostimulation. GnRH-I mRNA expression was determined using in situ hybridization in non-photostimulated (NPS), egg-laying (LAY), incubating (INC) and photorefractory (REF) hens. Overall, GnRH-I mRNA expression was greatest in the nucleus commissurae pallii (nCPa) and around the organum vasculosum lamina terminalis (OVLT), with less expression observed in the nucleus septalis lateralis (SL), cortico-habenula cortico-septum area, and within the nucleus preopticus medialis. GnRH-I mRNA expression was significantly increased in nCPa, OVLT, and SL after NPS hens (6L:18D) were exposed to a 30 or 90 min pulse of light beginning 14 h after first light (dawn). GnRH-I mRNA abundance within nCPa, OVLT and SL was greater in LAY than in NPS and INC hens, while mRNA expression was least in REF hens. These results indicate that GnRH-I mRNA expression in birds is sensitive to light stimulation during the photosensitive period and can be used to more precisely characterize their different reproductive stages.

In mammals, prolactin (PRL) secretion is regulated by vasoactive intestinal peptide (VIP) and peptide histidine isoleucine (PHI). In birds, however, VIP is considered a PRL-releasing factor (PRF), while the role of PHI is unknown. The purpose of this study was to compare the effects of turkey PHI (tPHI) and turkey VIP (tVIP) on PRL secretion in vitro, and to study their physiological significance in vivo through active immunization against tPHI and tVIP. In vitro studies were conducted using pituitary cell cultures from female turkeys. In the in vivo study, female turkeys were immunized with keyhole limpet hemocyanin (KLH; control), synthetic tPHI conjugate (KLH-tPHI), or synthetic tVIP conjugate (KLH-tVIP). Both tVIP and tPHI stimulated PRL secretion from anterior pituitary cells in a dose response manner. However, tPHI was 100-fold less potent than tVIP in stimulating maximum PRL secretion in vitro. In addition, the highest dose (10(-4) M) of tPHI inhibited its own PRL-releasing activity as well as that of VIP-stimulated PRL release. Whereas, circulating PRL levels and nesting activity remained low and unchanged during the photo-induced reproductive cycle (i.e., experimental period) in tVIP-immunized birds, control and tPHI-immunized turkeys showed a significant increase in plasma PRL levels and in the incidence of incubation behavior over time following photostimulation. These findings, taken together with earlier results, indicate that VIP is the sole physiological PRF in the turkey (avian species).

Although the actions of heroin on central nervous system (CNS) development are mediated through opioid receptors, the net effects converge on dysfunction of cholinergic systems. We explored the mechanisms underlying neurobehavioral deficits in mouse and avian (chick, Cayuga duck) models. In mice, prenatal heroin exposure (10 mg/kg on gestation days 9-18) elicited deficits in behaviors related to hippocampal cholinergic innervation, characterized by concomitant pre- and postsynaptic hyperactivity, but ending in a reduction of basal levels of protein kinase C (PKC) isoforms betaII and gamma and their desensitization to cholinergic receptor-induced activation. PKCalpha, which is not involved in the behaviors studied, was unaffected. Because mammalian models possess inherent confounding factors from maternal effects, we conducted parallel studies using avian embryos, evaluating hyperstriatal nucleus (intermedial part of the hyperstriatum ventrale, IMHV)-related, filial imprinting behavior. Heroin injection to the eggs (20 mg/kg) on incubation days 0 and 5 diminished the post-hatch imprinting ability and reduced PKCg and bII content in the IMHV membrane fraction. Two otherwise unrelated agents that converge on cholinergic systems, chlorpyrifos and nicotine, elicited the same spectrum of effects on PKC isoforms and imprinting but had more robust actions. Pharmacological characterization also excluded direct effects of opioid receptors on the expression of imprinting; instead, it indicated participation of serotonergic innervation. The avian models can provide rapid screening of neuroteratogens, exploration of common mechanisms of behavioral disruption, and the potential design of therapies to reverse neurobehavioral deficits.

Reproductive failure associated with heat stress is a wellknown phenomenon in avian species. Increased prolactin (PRL) levels in response to heat stress have been suggested as a mechanism involved in this reproductive malfunction. To test this hypothesis, laying female turkeys were subjected to 408C for 12 h during the photo-phase daily or maintained at 24‐268C. Birds in each group received oral treatment with parachlorophenyalanine (PCPA; 50 mg/kg BW/day for 3 days), an inhibitor of serotonin (5-HT) biosynthesis, or immunized against vasoactive intestinal peptide (VIP). Both treatments are known to reduce circulating PRL levels. Nontreated birds were included as controls. In the control group, high ambient temperature terminated egg laying, induced ovarian regression, reduced plasma luteinizing hormone (LH) and ovarian steroids (progesterone, testosterone, estradiol) levels, and increased plasma PRL levels and the incidence of incubation behavior. Pretreatment with PCPA reduced (P , 0.05) heat stress-induced decline in egg production, increase in PRL levels, and expression of incubation behavior. Plasma LH and ovarian steroid levels of heat stressed birds were restored to that of controls by PCPA treatment. As in PCPA-treated birds, VIP immunoneutralization of heat-stressed turkeys reduced (P , 0.05) circulating PRL levels and prevented the expression of incubation behavior. But it did not restore the decline in LH, ovarian steroids, and egg production (P . 0.05). The present findings indicate that the detrimental effect of high temperature on reproductive performance may not be related to the elevated PRL levels in heat-stressed birds but to mechanism(s) that involve 5-HT neurotransmission and the induction of hyperthermia. birds, egg production, heat stress, incubation behavior, luteinizing hormone, ovary, pituitary hormones, prolactin, reproduction, reproductive hormones, serotonin, stress, turkeys, vasoactive intestinal peptide

Photostimulation with green light accelerated BW and muscle development of broilers. In experiment 1, temperature sensors were inserted into 50 broiler eggs. The eggs were placed under 5 green light-emitting diode (LED) lamps at an intensity of 0.1 W/m2 at eggshell level for 5, 10, 15, 20, and 25 min (n = 10). Egg temperatures were recorded continuously. A high correlation was found between lighting period and egg temperature elevation, and an intermittent light regimen of 15 min on and 15 min off was found to eliminate light-induced egg overheating. In experiment 2, the effect of in ovo green light photostimulation on embryonic development was studied. Five hundred fertile eggs were divided into 2 groups: the first was photostimulated with green light from 5 d of incubation until hatch (0.1 W/m2 intensity) and the second was incubated in the dark. In ovo green light photostimulation caused a significant elevation in BW and breast muscle weight during embryo development and posthatch until 6 d of age. In experiment 3, 240 fertile broiler eggs were divided into 2 groups as described in experiment 2. At hatch, chicks from each in ovo light treatment were divided into 2 subgroups: the first was reared under green light and the second under white light. In ovo photostimulation with green light enhanced BW and breast muscle weight. However, rearing under green light did not have any synergistic effect on BW. Collectively, the results suggest that stimulation with green light enhances development and growth in chicks and that the best effect is achieved when this stimulus is provided during incubation.

Previous reports have suggested that green light enhances broiler growth at an early age, whereas blue light enhances growth at older ages. The aim of this study was to examine the effect of a switch in monochromatic light at 2 ages on growth and development of broilers. Male chicks (Anak, n = 640) were used. After hatch, chicks were weighed, wing-banded, and blocked into treatment groups. Chicks were grown in 1-m2 pens in 8 isolated light-proof rooms (20 birds/pen). The light treatments were (1) Control white (mini-incandescent lamps), 2) blue light-emitting diode (LED) lamps, 3) green LED lamps, 4) blue LED switching to green at 10 d of age, 5) blue LED switching to green at 20 d of age, 6) green LED switching to blue at 10 d of age, and 7) green LED switching to blue at 20 d of age. There were 8 pens for treatment 1, and 4 pens for each of the other treatments. The light schedule was 23L:1D, and intensity was 0.1 watts/m2. BW and feed consumption were recorded. Green light birds were significantly heavier at 4 d of age. Switching light at 10 d of age from green to blue caused a further increase in BW. This improved growth was maintained until the end of the experiment. Light switching from blue to green at 20 d of age also improved growth as compared with white light. Average feed efficiency and mortality rate did not differ between groups. No association was observed among light treatment, performance, and plasma triiodothyronine concentration. We suggest that green light stimulated growth of birds at early age, and shifting birds to a different light environment at 10 or 20 d of age might further stimulate growth.

Reduction in reproductive potential of ganders with progress in seasonal breeding is a known problem in commercial geese production. The role of changes in hypothalamic-pituitary-gondal axis and testis functions in this process is not clear. This article presents studies on the hypothalamic (GnRH-I, vasoactive intestinal peptide), pituitary (LHβ, prolactin [PRL], PRL receptor [PRLR]), testis (PRLR) axis messenger RNA (mRNA) expression during different stages of the breeding period and photoperiodic conditions. Testis mass; histologic and functional (testosterone [T]) parameters; and plasma concentrations of T, LH, and PRL were evaluated. We collected (six times) samples from 2-year-old ganders (n = 48) maintained in short day (10L:14D) during the period from November to July. Moreover, in the peak of sexual activity (March), an additional group was on exposure (6 weeks) to long day (LD; 16L:8D). During the first half of reproduction (January, March; photosensitive period), GnRH-I (1.9 vs. 0.3 relative quantity [RQ]) and LHβ (3.0 vs. 0.7 RQ) mRNA transcript expression and concentrations of T (1.9-2.9 vs. 0.3 ng/mL), LH (13.6-7.4 vs. 0.7 ng/mL) were found to be higher (P < 0.05) than at the end of breeding (July). With progress in breeding, marked elevation (P < 0.05) in PRL (22.0-387.1 ng/mL) concentration related to similar changes in vasoactive intestinal peptide (0.9-3.0 RQ) and PRL mRNA abundance (1.3-11.5 RQ; May, July) was observed. However, testis PRLR mRNA increased (P < 0.05) only at the end of reproduction (1.2 RQ) compared to the peak of sexual activity (0.4 RQ; March). Furthermore, changes in mRNA transcript expression of the lactotrophic axis were accompanied with reduction of testis weight (left: 11.1-5.8 g), spermatogenesis (spermatogenic index: 5.4-3.0), and steroidogenesis (T: 24.8-1.3 ng/g testis), which may suggest their pivotal inhibitory modulation role in the regression of seasonal reproductive activity in ganders. The LD conditions (similar to spring-summer) resulted in earlier peripheral changes in T (0.9 vs. 1.8 ng/mL), LH (1.1 vs. 3.8 ng/mL), and PRL (296.1 vs. 161.2 ng/mL) concentrations than in short day, and this may be related to the advance in the timing of the sexual activity failure observed under natural light regimes. The lack of differences in gonadal and lactotrophic axis mRNA expression after LD treatment suggested a regulation based on the posttranslational mechanisms or modification of transcript or protein.

Melatonin (MLT) has a significant role in mammalian reproduction, with little or no effect in birds. In the present study we studied the role of MLT in regulation of luteinizing hormone (LH) secretion in castrated White Leghorn (WL) roosters. In Experiment 1, castrated WL roosters (n = 30) were divided into three groups, and each group (n = 10) was subdivided into two subgroups (n = 5). Birds in one subgroup received an injection of MLT at 5 mg (MLT-5), 20 mg (MLT-20), or 80 mg (MLT-80)/kg BW. Birds in the second subgroup were vehicle-injected and served as controls. Each dose of MLT was administered on a separate day at 1100 h. Blood was sampled 30 min before and 10, 30, 60, and 120 min after MLT or vehicle administration. Ten minutes after MLT administration, a significant reduction in plasma LH was observed in the MLT-20 and -80 groups, i.e., 70.3 +/- 8.3% and 62.2 +/- 4.1% of control values, respectively. In the MLT-80 group, plasma LH further declined to 42.1 +/- 9.7% of control values 60 min after injection. In Experiment 2, 18 castrated WL roosters were divided into three groups of six birds each. Two groups were injected with 80 mg MLT/kg BW at the beginning of the experiment; the second group received an additional dose of 80 mg MLT/ kg BW 140 min after the first injection. The third group was injected twice (as in second group) with vehicle and served as control. Blood was sampled 30 min before and 30, 60, 120, 170, 200, and 240 min after injection. Repeated MLT injection maintained low levels of plasma LH level until the end of the experiment. In Experiment 3, 10 castrated WL roosters were divided into two groups (n = 5). The first group was injected daily, for 10 d, with 80 mg MLT/kg BW, the second group was vehicle-treated and served as a control. At Day 3, there was a significant reduction in plasma LH level in the MLT-treated group, which continued for 7 d. This study demonstrates that, in birds, MLT suppresses LH secretion in a dose- and a time-related manner.

In flocks of geese fertility decreases in the second half of the breeding season. The reasons for this reduction in reproduction ability are still unclear. This study measured changes in semen quality variables throughout the period of intensive breeding in relation to hormonal concentrations associated with the sexual activity of ganders. Semen was collected (2×/week) from 2-year-old ganders in the period February–June. Standard ejaculation parameters and spermatozoa (spz) membrane integrity after E/N and SYBR-14/PI staining were evaluated. The DNA Fragmentation Index was measured by flow cytometry and sperm quality factors (SQF). The plasma levels of T, E2, P4, LH, PRL, THs in relation to semen parameters were evaluated. In ejaculate collected at the onset of the second half of breeding (April – spring period), a reduction in sperm concentration and % of live E/N and live SYBR-14+/PI− spz was shown. At this time, decrease in concentrations of LH and T and increase in PRL were found as well as moderate changes in THs were observed. However, in May a second peak in T and sperm production occurred. The DFI-% was higher in the middle part of breeding. Gonadal steroids concentration were not good prognostic marker of the reproductive potential of ganders. We suggest that a marked decline in LH and T in the spring period indicated the onset of endocrine changes mediated by PRL and THs resulting in progressive regression of testis functions. The lowest SQF in the spring/summer period coincided with the highest PRL suggesting an anti-spermatogenic action of PRL in ganders.

The present studies employ multitudinous approaches in order to overcome the methodological obstacles in the understanding of the relationship between neurochemical alterations and behavioral deficits induced by heroin during prenatal development. Mice were exposed prenatally to heroin via daily subcutaneous injections of 10 mg/kg, on gestation days 9-18. At age 50 days, the heroin-exposed offspring displayed behavioral deficits as assessed in the eight-arm and Morris mazes, pointing to possible alteration in the septohippocampal cholinergic innervations. Biochemically there was increased presynaptic activity of these innervations as attested to by the increased [3H]hemicholinium-3 (HC-3) binding sites and by K+-stimulated inositol phosphate (IP) formation. Postsynaptically, there was global hyperactivation along the different components of the nerve conduction cascade, including an increase in M1 muscarinic receptor Bmax, a general increase in G-proteins (GP) including the most relevant, G subtype, and an increase in IP formation and in basal protein kinase C (PKC) activity. However, there was desensitization of PKC activity in response to cholinergic agonist in the heroin-exposed offspring. Transplantation of normal embryonic cholinergic cells to the impaired hippocampus reversed the behavioral deficits and both the pre- and postsynaptic hyperactivity and resensitized PKC activity. To support and further strengthen the findings of the neural grafting study, correlation of the heroin-induced behavioral deficits with the biochemical alterations, done within individuals, was applied. The results showed high r values for IP formation, basal PKC, and PKC desensitization. The r values for HC-3 binding were statistically significant but relatively low. Taken together, the findings of the neural grafting and correlation studies bring us closer to understanding the relationship between the prenatal heroin-induced biochemical and behavioral changes. However, mammalian models possess the inherent methodological hindrances, stemming from possible maternal effects. To provide a control for these confounding variables, a chick embryo model was applied in which filial imprinting, a behavior related to a specific hyperstriatal nucleus, served as an endpoint. Heroin was administered to developing chick embryos by injecting the eggs (20 mg/kg) on incubation days (ID) 0 or 5. Prehatch exposure to heroin markedly diminished the ability for filial imprinting in the hatched chicks.

1. The effect of vitamin A on the small intestine was examined in vitamin-A-deficient meat-type chickens. 2. Maturation and activity of the small intestinal cells were assayed by detection of proliferating cells with proliferating cells nuclear antigen, goblet cells with Alcian blue, mature cells with alkaline phosphatase and extent of RNA expression with dot blot analysis. 3. Vitamin A deficiency caused hyperproliferation of enterocytes, a decrease in the number of goblet cells, decreased alkaline phosphatase activity and decreased expression of 2 brush-border enzymes. 4. Our findings suggest that the absence of vitamin A interferes with the normal growth rate in chickens because it influences functionality of the small intestine by altering proliferation and maturation of cells in the small intestinal mucosa.

This study investigated the effect of feeding in hatching trays on incubator weight loss and posthatch growth of chicks and poults. After hatch body weight (BW) of poultry decreased linearly in the hatching trays between 0.14 and 0.17 g/hr. When feed was present, BW loss was initially linear and then plateaued. Chicks and poults with access to feed were 2 to 3.5g heavier, when removed from the incubator, than were held birds. Egg weight was correlated with hatch weight, but not with time of hatching. The enhanced BW at placement due to feed ingestion in the hatchery was maintained through 21 days, with early hatching chicks and poults showing the greatest hatchery decrease in BW (partially alleviated by feeding). Late hatching poultry were less affected by holding or feeding. Availability of feed in hatching trays decreased BW losses in the hatchery and enhanced posthatch growth.

Inherited overfeeding and fattiness reduce laying performance in broiler breeder pullets. Although feed restriction is used to compensate for overeating and weight gain, this management practice leads to increases in BW variation, labor cost, and bird stress. Dietary supplementation of anorectic agents, such as fenfluramine, may be an alternative. Anak female prebreeder hens (19 wk of age; n = 10 per group) were treated as follows: daily oral administration of 5, 10, 20, or 40 mg DL-fenfluramine/kg BW or saline with food provided for ad libitum intake or administration of saline and feed restriction. Daily feed intake (FI), laying rate, egg composition, and BW were measured. At 40 wk of age, adipose tissue and ovary weights were measured. Fenfluramine depressed (P0.05) BW and FI in a dose-dependent manner, but was less effective in reducing BW than feed restriction. Suppression of FI occurred in two phases: a dynamic phase, coinciding with the rapid growth phase, during which FI declined progressively and a static phase during which FI reached a plateau at a significantly low level until the end of the experimental period. Egg production peaked first in saline-treated hens fed for ad libitum intake, but soon after started to decline. In all fenfluramine-treated and feed-restricted hens, egg production peaked 3 to 4 wk later and remained high until the end of the experiment. There were no differences in egg and egg component weights among the experimental groups. Abdominal adipose tissue weight was reduced by fenfluramine in a dose-related manner, and its weight in the group treated with the highest dose was similar to that of feed-restricted hens. In these two groups, ovarian weight was significantly higher than in the saline-treated hens fed for ad libitum intake, and a small, nonsignificant increase in ovary size was observed in groups treated with the two median doses of fenfluramine. The effect of fenfluramine on egg production was similar to that of feed restriction, but it was not dose-dependent and, thus, not directly related to its leaning effect. In broiler breeder hens, oral fenfluramine may be used for chemical feed restriction and diminution of fattiness without reducing egg production relative to manually feed-restricted hens.

The relationships between testicular and plasma hormone levels and the decline in fertility in aging roosters were examined. Body mass, testicular mass, and fertility were measured in roosters from 20 to 72 weeks of age. Plasma was assayed for LH and testosterone, and estradiol and testicular extracts were assayed for testosterone and estradiol contents. Fertility increased rapidly in young roosters to a peak of 96.2 +/- 3.9% at 37 weeks of age. Thereafter, fertility declined and by 72 weeks of age was significantly lower than at 37 weeks. Plasma LH reached 16.8 +/- 2.5 ng/ml at 27 weeks and remained high until 60 weeks of age, when it decreased significantly. Plasma and testicular testosterone levels increased from low levels in young birds to a peak that coincided with highest fertility and declined thereafter. Plasma and testicular estradiol showed a striking inverse relationship with testosterone. Plasma estradiol was 29.4 +/- 4.0 pg/ml in 20-week-old birds, decreased rapidly as testosterone increased, and increased again in older birds as testosterone decreased. Thus, the decline in fertility in aging roosters was associated with a decrease in plasma LH and testosterone and an increase in plasma and testicular estradiol. It is suggested that plasma levels of LH and testosterone in roosters are regulated by a negative feedback mechanism involving estradiol that is produced not only by the aromatization of testosterone in the brain but also by peripheral estradiol originating in the testes and that estradiol has a major role in the decline in fertility in aging roosters.

Artificial illumination, including light quality, is crucial in modem broiler management. In the present study, a new, highly efficient, monochromatic light system has been developed for broilers. One hundred and eighty male broiler chicks (Anak) were divided into four light treatment groups (n = 45) in three replicates each. All birds were housed in a single room previously divided by wooden bars into 12 sealed cells of 1 m2. Feed and water were provided for ad libitum consumption. Light intensity was 0.1 W/m2 at the height of birds' heads and was scheduled for 23 h of light and 1 h of dark during the entire experimental period. Light treatments were: control white (mini-incandescent light bulbs), blue (480 nm), green (560 nm), and red (660 nm). Body weight was recorded periodically, feed consumption was measured daily, and feed efficiency was calculated. Blood samples were taken at 1, 9, and 32 d of age and plasma testosterone was determined. Two necropsies were conducted, at 23 and 35 d of age, and selected glands and organs were weighed. In the group reared under green light, a significant enhancement in weight gain was observed as early as 3 d of age; this gain was maintained during the entire experimental period. Broilers reared under blue light had a later onset of growth enhancement and were significantly heavier than those reared under white and red light at 20 d of age. Plasma testosterone levels were significantly higher in birds reared under blue light. Breast muscle weights were significantly higher in the birds reared under green light at 23 and 35 d of age. These results suggest that green and blue light stimulate growth.

Artificial illumination is an important factor in the management of layers. In this study, a new monochromatic light system was developed for egg layers. Prelaying pullets (Lohmann) were marked and housed in nine light and temperature control rooms (15 battery cages, 3 hens per cage; n = 45), divided into three light treatments: 0.1 and 0.01 W/m2 light intensity using light emitting diode (LED) lamps and 0.1 W/m2 using mini-fluorescent bulbs (PL) (control). In each of the LED rooms, three wavelengths were tested: 560 (n = 9), 660 (n = 9), 880 (n = 6), and 660 intermitted lighting (15 min light 45 min dark, 660IN) (n = 9). Birds were exposed to 12 h light and 12 h of darkness using PL lamps. At 21 wk of age, the light period was increased to 12.75 h by using 5.5 h of LED lamps and 7.25 of PL light source for Groups 1 and 2, the third group received 12.75 h of PL light. Until 28 wk of age, light hours increased by 0.5 h/w using LED light for Groups 1 and 2 and PL source for the third group, reaching 16 h of light at 28 wk of age. Egg production and feed consumption were recorded daily; egg components were recorded weekly for 10 mo. A significant reduction in egg production was observed in all 880nm groups; no differences in egg production and quality were found in the other groups. Feed consumption was significantly lower by 7% in all 0.01 W/m2 groups. We suggest that an important reduction in rearing costs of laying hens may be obtained by using this system.

The involvement of melatonin (Mel) in body temperature (Tb) regulation was studied in White Leghorn layers. In experiment 1, 35 hens were injected intraperitoneally with seven doses of Mel (0, 5, 10, 20, 40, 80, or 160 mg Mel/kg body wt) dissolved in ethanol. Within 1 h, Mel had caused a dose-dependent reduction in Tb. To eliminate a possible vehicle effect, 0, 80, and 160 mg/kg body wt Mel dissolved in N-methyl-2-pyrrolidone (NMP) was injected. NMP had no effect on Tb, with Mel again causing a dose-dependent hypothermia. In experiment 2 ( n= 30), Mel injected before exposure of layers to heat reduced Tb and prevented heat-induced hyperthermia. Injection after heat stress had begun did not prevent hyperthermia. Under cold stress, Mel induced hypothermia, which was not observed in controls. In experiment 3 ( n= 12), Mel injection reduced Tband increased metatarsal and comb temperatures (but not feathered-skin temperature), respiratory rate, and evaporative water loss. Heart rate rose and then declined, and blood pressure increased 1 h after Mel injection. Heat production rose slightly during the first hour, then decreased in parallel to the Tbdecline. We conclude that pharmacological doses of Mel induce hypothermia in hens by increasing nonevaporative skin heat losses and slightly increasing respiratory evaporation.