Alterations in gonadotropin, prolactin, androgen and estrogen receptor and steroidogenesis-associated gene expression in gander testes in relation to the annual period.

Physiological mechanisms of seasonal changes in testicular function in birds are not fully elucidated. The balance between androgens and estrogens and testis sensitivity for gonadotropin and gonadal steroids are still unclear. The aim of the study was to examine: (1) the changes in circulating and intra-testicular steroid hormone levels and their relationship; (2) the mRNA expression of testicular gonadotropin, prolactin (PRL), progesterone (P4), androgen, and estrogen receptors, and (3) key steroidogenesis processes-related genes with immunofluorescent localization of aromatase in gander testes during the annual period. Testes from ganders (n = 25) in the first reproduction season were obtained at five breeding stages, i.e., prebreeding (PrB), peak of reproduction (PR), postbreeding (PoB), nonbreeding (NB), and onset of reproduction (OR). Males were kept under breeding conditions. It was found that plasma P4 levels decreased at the PoB and NB stages, whereas intra-testicular P4 was the highest in the NB stage. Intra-testicular estradiol (E2) levels were higher at the PoB and NB stages than the other stages, whereas testosterone (T) levels showed a nearly opposite pattern. The plasma estradiol-to-testosterone ratios were higher at the PrB, PoB and NB stages compared to other stages. The transcript abundances for luteinizing hormone receptor (LHR), PRL receptor (PRLR), estrogen receptor alpha (ERα), and estrogen receptor beta (ERß) also change in testicular tissue during the annual period. Moreover, StAR mRNA expression was upregulated at the PoB and NB stages, and CYP11A1 transcript level was the highest at the PoB stage. Stage-dependent changes in the CYP19A1 mRNA and aromatase protein levels with higher abundances of transcript at PoB and NB stages and protein at the NB stage were observed. Localization and immunofluorescent signal intensity for aromatase also differed in relation to the examined stages. It may be suggested that differential E2 levels, as well as aromatase expression and localization across annual stages are responsible for the seasonal activation/inactivation stages of testis spermatogenesis in domestic ganders. These data strongly suggest a role of aromatase in the control of gander steroidogenesis as changes in this enzyme level are associated with alternation in gonadal steroid hormones. In addition, joint action with others hormones, like PRL and LH, seems to be important in the final effect of seasonal reproduction potential.

Parental Preconception and Pre-Hatch Exposure to a Developmental Insult Alters Offspring's Gene Expression and Epigenetic Regulations: An Avian Model.

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.

Annual changes in cell proliferation and apoptosis and expression of connexin 43 in the testes of domestic seasonal breeding ganders.

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 (H&E 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.

Beta-glucans induce cellular immune training and changes in intestinal morphology in poultry.

Introduction: Beta-glucans are known as biological response modifiers due to their ability to activate the immune system. This research aimed to determine the efficacy and safety of feeding beta-glucans from various sources on the immune status and intestinal morphology of chickens. Methods: To this end we used in vitro and in vivo set-ups. In the in vitro set-up the chicken macrophage cell line HD-11 was used to measure the response of the chicken immune cells to beta-glucans extracted from algae and mushrooms on immune-related gene expression and associated activities. Additionally, we conducted two in vivo experiments using either beta-glucans extracted from yeast or mix of yeast and mushrooms beta-glucans as part of the chicks feed in order to test their effects on the chick intestinal morphology. Results: In the in vitro set-up exposure of HD-11 cells to a concentration of 1 mg/ml of algae and mushroom beta-glucans resulted in significantly higher expression of 6 genes (TNFα, IL4, IL6, IL8, IL10, and iNOS2) compared to control. The release of nitrite oxide (NO) to the medium after exposure of HD-11 cells to mushrooms or algae beta-glucans was significantly increased compared to control. Additionally, significantly increased phagocytosis activity was found after exposure of the cells to algae and mushroom beta-glucans. In the in vivo set-up we observed that the length of the villi and the number of goblet cells in the ileum and the jejunum in the beta-glucan fed chicks were significantly augmented compared to control, when the chicks were fed with either yeast or yeast and mushroom beta-glucans mix. Discussion: In conclusion, dietary supplementation of poultry with beta-glucan exerts significant and positive effects on immune activity and the intestinal morphology in poultry.

Determination of Annual Plasma Hormone Levels Associated with Reproduction in Long-Day Breeding Domestic Geese.

This paper examines the dynamics of circulating hormone changes connected with reproduction in geese during the annual period related to gonad morphometry. One hundred geese were examined. The levels of prolactin (PRL), triiodothyronine (T3), thyroxine (T4), testosterone (T), progesterone (P4) and estradiol (E2) were estimated. In both sexes, PRL level patterns fit a quadratic trend with elevations in the post-breeding and the second half of the breeding-laying periods. During these periods, differences in the PRL level between sexes were noted. In ganders, increased PRL levels during the laying period occurred earlier compared to in female geese. Cubic trends for T and E2 in ganders and quadratic for T, P4, and E2 in female geese were observed. PRL was negatively correlated with T in both sexes and with P4 and E2 in female geese. A higher level of T3 and variation in T4 in ganders with a quartic trend in ganders vs. a quadratic in female geese were noted. Patterns of PRL, T, and E2 suggested that the breeding-laying period in ganders may be shorter than in female geese. These findings will be used to explore experimental manipulations of the endocrine axis to increase synchronisation of both sexes.

Targeted differential monochromatic lighting improves broiler breeder reproductive performance.

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.

The effect of parachlorophenylalanine treatment on the activity of gonadal and lactotrophic axes in native Polish crested chickens stimulated to broodiness.

Brooding behavior, a common characteristic of native breeds of the domestic chicken, is marked by elevated prolactin (PRL) levels, which is necessary for incubation and connected with changes in hypothalamic-pituitary-gonadal axis activity. Evidence indicates the serotoninergic system is a potent modulator of PRL secretion. The objective of this study is to investigate whether blocking serotonin synthesis with parachlorophenylalanine (PCPA) prevents incubation behavior in native Polish crested chickens. In addition, we examined the effect of PCPA on the gene expression of the gonadal and lactotrophic axes. Birds were stimulated to broodiness by artificial eggs in nests. At 34 wk of age (April: spring period), the hens were divided into 2 groups (14 hens in each group): control and PCPA-treated (50 mg/kg BW) group. After 5 wk of treatment, the artificial eggs were removed from the nests. Egg production, incubation activity, and levels of plasma ovarian steroids progesterone (P4), testosterone (T), estradiol (E2), and PRL were examined. At the end of the experiment (45 wk of age, June: summer period), ovarian characteristics and mRNA gene expression of gonadal (gonadotropin-releasing hormone [GnRH] I, luteinizing hormone [LH] ß, follicle-stimulating hormone [FSH] ß) and lactotrophic (vasoactive intestinal peptide [VIP], PRL) axes were measured by quantitative real-time PCR. Incubation activity was observed in the hens of both groups but with lower frequency in PCPA-treated birds. Moreover, the PCPA group had a higher cumulative egg production than the controls. During the first six and 8 wk of the experiment, levels of P4 and E2, respectively, were similar in both groups, but all concentrations increased in the PCPA-treated hens after this period. In addition, increased GnRH-I, LHß, and FSHß and decreased VIP mRNA expression was observed in the PCPA group compared with the controls. There were no differences in PRL mRNA expression, the PRL level, and ovarian morphometry between the 2 groups. These results indicate that blockage of serotonin synthesis by PCPA does not effectively prevent incubation in native Polish crested chickens. However, treatment with PCPA increased gonadal axis activity and improved reproductive performance.

Hormonal levels of estradiol, testosterone, and progesterone at entry into lay of year 1980 vs. 2000 broiler breeder females under fast and slow release from feed restriction.

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 effects of replacing eggs with chicks on mesotocin, dopamine, and prolactin in the native Thai hen.

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.

Distribution of hypothalamic vasoactive intestinal peptide immunoreactive neurons in the male native Thai chicken.

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.

Mating activity and sperm penetration assay in prediction of the reproduction potential of domestic goose ganders in a harem system.

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.

Effect of breeding stage and photoperiod on gonadal and serotonergic axes in domestic ganders.

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 (LHß, 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 LHß (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.

Breeding period-associated changes in semen quality, concentrations of LH, PRL, gonadal steroid and thyroid hormones in domestic goose ganders (Anser anser f. domesticus).

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 liveE/N and liveSYBR-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.

Dopamine and prolactin involvement in the maternal care of chicks in the native Thai hen (Gallus domesticus).

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<0.05). However, there was no significant change in the DAergic activity in the ML in either the R or NR groups throughout the 28-day rearing periods. These results suggest that the DA/PRL system is involved in early rearing behavior. The additional decline in DAergic activity and plasma PRL levels during the disruption of rearing behavior further supports their involvement in rearing behavior in this equatorial precocial species.

Mating activity of domestic geese ganders (Anser anser f. domesticus) during breeding period in relation to age, testosterone and thyroid hormones.

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.

Temporal activity and dietary selection in two coexisting desert snakes, the Saharan sand viper (Cerastes vipera) and the crowned leafnose (Lytorhynchus diadema).

The Saharan sand viper (Cerastes vipera) and the crowned leafnose (Lytorhynchus diadema) are two snake species well adapted to desert sand dunes and, in Israel, coexist in the western Negev Desert. C. vipera is a sit-and-wait ambusher while L. diadema is an active hunter. We studied the seasonal and diel activity patterns and dietary selection of these two species while free-living in the field. Both species were active from early spring until late fall but displayed (i) different seasonal activity patterns -C. vipera was bimodal with peaks in spring and autumn whereas L. diadema was basically unimodal with a peak in summer; (ii) different nocturnal above-ground activity patterns -C. vipera was active mainly during the first three hours of darkness while L. diadema was constantly active during the first seven hours of darkness; and (iii) different patterns of nocturnal behavior -C. vipera moved up to 50 m while L. diadema moved several hundred meters each night. Dietary selection differed between these snake species although lizards, mainly Nidua fringe-fingered lizards (Acanthodactylus scutellatus), were the main dietary item for both. We concluded that temporal partitioning in above-ground activity, different foraging strategies and differences in dietary selection may contribute towards the coexistence of the two snake species.

The effect of parachlorophenylalanine and active immunization against vasoactive intestinal peptide on reproductive activities of broiler breeder hens photostimulated with green light.

Photostimulation of retinal photoreceptors appears to inhibit reproductive activity in birds. In the present study, the involvement of serotonin and vasoactive intestinal peptide was investigated in relation to reproductive failure associated with retinal photostimulation. Hens at 23 wk of age were divided into six rooms equipped with individual cages. At 24 wk of age, three rooms were photostimulated (14L:10D) with white light (control). Three rooms had two parallel lighting systems, red (660 nm) and green (560 nm), which were both on during 6 h of the 14-h light period. Upon photostimulation, the red light was turned off after 6 h, and the green light was left on for a total of 14 h (Green). Five hens from each room served as controls, five hens were immunized against vasoactive intestinal peptide, and five hens received parachlorophenylalanine, an inhibitor of serotonin biosynthesis. Parachlorophenylalanine treatment increased reproductive performance and mRNA expression of GnRH-I, LH-beta and FSH-beta (P < 0.05) in the Green group to levels which did not differ from those of the White (control) group. Immunization against vasoactive intestinal peptide reduced plasma concentration and pituitary mRNA expression of prolactin but did not affect expression of gonadal axis genes. Collectively, the results suggest that retinal photostimulation inhibits the reproductive axis through serotonin and not through vasoactive intestinal peptide.

Neuroendocrine regulation of rearing behavior in the native Thai hen.

Vasoactive intestinal peptide (VIP) is the avian prolactin releasing factor and changes in the concentrations of plasma prolactin (PRL) are found during the avian reproductive cycle. This study investigated the changes in the VIP/PRL system of native Thai hens rearing their young as compared to hens deprived of rearing their chicks. The number of VIP-immunoreactive (VIP-ir) neurons in the Nucleus inferioris hypothalami (IH) and Nucleus infundibuli hypothalami (IN) of hens rearing chicks (R) were compared with those of non-rearing chicks (NR). Plasma PRL levels were determined by enzyme-linked immunosorbent assay. The localization and number of VIP-ir neurons were determined by immunohistochemistry. The numbers of VIP-ir neurons in the IH-IN areas were high in the R hens, whereas the number of VIP-ir neurons decreased in the NR hens as compared to their respective R hens. During the rearing period, changes in the VIP-ir neurons within the IH-IN were correlated with plasma PRL levels. The results of the present study indicate for the first time that the VIP/PRL system plays a role in neuroendocrine reorganization to establish maternal behavior in native Thai chickens. The VIP/PRL system functions not only as a well established key regulator of incubation behavior, but is also involved in the regulation of rearing behavior. It is possible that VIP and the decline in the number of VIP-ir neurons and in turn VIPergic activity and the decrease in PRL levels are related to their contribution to rearing behavior of this non-seasonal breeding, equatorial precocial species.

Reproductive cycle of free-living male Saharan sand vipers, Cerastes vipera (Viperidae) in the Negev desert, Israel.

The Saharan sand viper, Cerastes vipera (Linnaeus, 1758), is distributed in all Saharan countries, being confined to sand and dune systems. This relatively small snake, up to 35 cm, is nocturnal, is active from spring to autumn (April to October) and hibernates during the winter (November to March). We predicted that C. vipera would have peak plasma testosterone concentration at mating and that the vas deferens would contain abundant spermatozoa at that time. To test our predictions, we collected information on the time of mating and measured monthly testosterone concentration, testes size and testicular activity in free-living male C. vipera during its active period from April to October. Mating occurred only during spring. The pattern of plasma testosterone concentration, testes volume, seminiferous tubule diameter and spermatogenesis all followed the general pattern of high values in autumn and spring and low values in early summer. Our predictions were partially supported. There was a high plasma testosterone concentration at mating in spring and the vas deferens contained abundant spermatozoa, as predicted, but there was also a high plasma testosterone concentration in autumn without mating. We concluded that: (1) males are both aestival in that they produce spermatozoa in autumn, which they store over the winter hibernation period, and vernal in that they produce spermatozoa in spring prior to mating; (2) matings are associated with spermatogenesis; and (3) the high plasma testosterone concentration is concomitant with both matings and spermatogenesis in spring and with spermatogenesis in autumn. We propose that C. vipera has a single peak of testicular activity and plasma testosterone concentration which start in autumn and end in spring. We also propose that spermatogenesis is prior to spring mating and, consequently, is prenuptial.

Changes in vasoactive intestinal peptide and tyrosine hydroxylase immunoreactivity in the brain of nest-deprived native Thai hen.

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.