Genetic architecture and genomic selection of female reproduction traits in rainbow trout.

BACKGROUND: Rainbow trout is a significant fish farming species under temperate climates. Female reproduction traits play an important role in the economy of breeding companies with the sale of fertilized eggs. The objectives of this study are threefold: to estimate the genetic parameters of female reproduction traits, to determine the genetic architecture of these traits by the identification of quantitative trait loci (QTL), and to assess the expected efficiency of a pedigree-based selection (BLUP) or genomic selection for these traits. RESULTS: A pedigreed population of 1343 trout were genotyped for 57,000 SNP markers and phenotyped for seven traits at 2 years of age: spawning date, female body weight before and after spawning, the spawn weight and the egg number of the spawn, the egg average weight and average diameter. Genetic parameters were estimated in multi-trait linear animal models. Heritability estimates were moderate, varying from 0.27 to 0.44. The female body weight was not genetically correlated to any of the reproduction traits. Spawn weight showed strong and favourable genetic correlation with the number of eggs in the spawn and individual egg size traits, but the egg number was uncorrelated to the egg size traits. The genome-wide association studies showed that all traits were very polygenic since less than 10% of the genetic variance was explained by the cumulative effects of the QTLs: for any trait, only 2 to 4 QTLs were detected that explained in-between 1 and 3% of the genetic variance. Genomic selection based on a reference population of only one thousand individuals related to candidates would improve the efficiency of BLUP selection from 16 to 37% depending on traits. CONCLUSIONS: Our genetic parameter estimates made unlikely the hypothesis that selection for growth could induce any indirect improvement for female reproduction traits. It is thus important to consider direct selection for spawn weight for improving egg production traits in rainbow trout breeding programs. Due to the low proportion of genetic variance explained by the few QTLs detected for each reproduction traits, marker assisted selection cannot be effective. However genomic selection would allow significant gains of accuracy compared to pedigree-based selection.

Behavioral and adrenal responses to various stressors in mule ducks from different commercial genetic selection schemes and their respective parental genotypes.

The mule duck, a hybrid produced by crossing a Muscovy drake and a Pekin female, is reported to express inappropriate behavior such as collective avoidance of people, the resulting distress and physical consequences potentially compromising their welfare. The present study was carried out to characterize the responses of mule duck strains from different commercial selection schemes to various stressful conditions and to confirm previous data on the genetic cross effects observed in a specific genotype. Three independent experiments were conducted with ducks from 3 French breeding companies (A, B, and C). Each experiment compared 2 mule genotypes sharing one common parental origin (paternal for ducks from company A or maternal for ducks from companies B and C). Mule duck males from the 2 genotypes and their respective parental genotypes (Pekin and Muscovy) were subjected to a set of social and stressful physiological and behavioral tests. Previously reported differences in genetic cross effects on fear responses between the parental genotypes and the corresponding hybrid were confirmed in these commercial crosses. Both mule duck and Pekin genotypes showed more active physiological and behavioral responses to stress than Muscovy genotypes. The new finding of this study is that mule genotypes appear to be more sensitive to the social environment than both respective parental genotypes. Few differences were observed between the 2 mule genotypes from A and C. On the other hand, several traits of the 2 mule genotypes from B differed. In addition, A and C mule genotypes were characterized by the same adrenal and behavioral traits but contrasting responses. The B mule genotypes were characterized by a different set of behavioral traits, and only 1 of the 2 B mule ducks was characterized by a group of adrenal traits.

Circulating corticosterone reaction to restraint and adrenocorticotropin hormone administration in white leghorns selected for immune response traits.

Corticosterone plasma concentration was measured in a random-bred control line and in 3 White Leghorn chicken lines previously selected over 9 generations for 3 different in vivo immune responses: high antibody response to Newcastle disease virus vaccine 3 wk after vaccination (ND3), high cell-mediated immune response (response to phytohemagglutinin, PHA), and high phagocytic activity measured as carbon clearance (CC). The objective of the study was to estimate if selection on immune response had an effect on the response to stress assessed by measures of corticosterone concentration before and after physical stress or adrenocorticotropin hormone injection and if the effect was dependent on the immune response trait that had been selected for, by joint analyses of immune responses and concentrations. The mean values of plasma corticosterone measures did not differ between lines, indicating that selection for different high immune responses had little effect on response to stress. Within line, however, significant negative correlations (-0.46

Relationship between hypothalamic-pituitary-adrenal axis responsiveness and age, sexual maturity status, and sex in Japanese quail selected for long or short duration of tonic immobility.

The influences of age (4 to 12 wk), sexual maturity status, and sex on hypothalamic-pituitary-adrenal (HPA) axis responsiveness were investigated by measuring changes in peripheral basal levels of corticosterone (B) and responses to 10 min of physical restraint in a crush cage or injection of 1-24 adrenocorticotropic hormone (ACTH) in 2 genotypes of Japanese quail divergently selected for long (LTI) or short (STI) duration of tonic immobility (TI). Although gonad development was more advanced in male STI quail, most birds were still immature at 4 wk of age, but sexual maturity was fully acquired by 6 wk of age in both sexes and genotypes. This change was associated with increases in basal B levels in both genotypes and sexes. On the other hand, HPA axis responsiveness to restraint and adrenal responsiveness to 1-24 ACTH injection remained stable in STI quail, whatever the age. Conversely, significant responses to restraint compared with basal B levels were only observed at 4 and 6 wk of age, and adrenal responsiveness increased with age in LTI quail. Moreover, higher B levels were measured in response to restraint in STI than in LTI quail, whereas similar adrenal responses were measured at 9 and 12 wk of age. We concluded that an increase in basal B levels is associated with the stage of sexual maturity acquired, but it did not affect HPA axis responsiveness or adrenal B response capacity. On the other hand, age affected HPA axis responsiveness and adrenal B response capacity in LTI quail of both sexes but not in STI quail. It is hypothesized that functional HPA axis maturity occurs after 6 wk of age in the LTI genotype, but it is reached by 4 wk of age in the STI genotype. In conclusion, the divergent selection program for TI conducted on quail resulted in changes in HPA responsiveness that probably resulted from differences in development rate and function of the adrenal glands or other upstream structures of the HPA axis.

Daily and photoperiod variations of hypothalamic-pituitary-adrenal axis responsiveness in Japanese quail selected for short or long tonic immobility.

The aims of this study were to investigate the existence of a circadian rhythm of basal corticosterone (B) plasma concentrations in male and female Japanese quail lines divergently selected for long (LTI) or short (STI) duration of tonic immobility (TI) and the possible effects of photoperiod length on corticotropic axis reactivity. Significant peaks in B levels were observed throughout the day in 3 out of the 4 groups used in our experiments. However, B levels remained very low for all groups (< 5.0 ng/mL) and there was no consensus between groups. We therefore have no evidence from our results that basal B levels follow a circadian rhythm in adult STI and LTI quail held under a long photoperiod (16L:8D). We also showed that rearing under a long photoperiod (16L:8D) was associated with higher basal B levels and higher B adrenal response capacity to 1-24 adrenocorticotropic hormone (ACTH) injection in the STI and LTI lines compared with a shorter period (8L:16D). Higher hypothalamic-pituitary-adrenal (HPA) axis responsiveness to restraint in a crush cage was also measured in female quail reared under the long photoperiod, and similar responses were measured under both photoperiods in males. This result suggests that the effects of photoperiod length involve both local and more central mechanisms in the control of HPA axis responsiveness according to sex. On the other hand, we showed that the genetic selection program for TI responses induced greater increases in the B level following restraint in STI quail than in LTI quail of both sexes under both photoperiods, but the B adrenal response capacity was similar for both lines and sexes. Although further investigations on both lines regarding adrenal sensitivity are necessary before being able to conclude definitively, our findings strongly suggest that the differences observed in HPA axis responsiveness to restraint between lines are probably not due to differences in adrenal function itself but may involve upstream structures of the HPA axis.

In-vitro and in-vivo responses to chicken LHRH-I and chicken LHRH-II in male turkeys (Meleagris gallopavo).

Stimulation of male turkey hypophyses in vitro with chicken (c)LHRH-I, cLHRH-II or porcine (p)LHRH (0.1 mumol/l) using a perifusion technique caused an increase in the release of LH. In this system, cLHRH-II was approximately 2.5-fold more potent than cLHRH-I and pLHRH which were equipotent. The difference was due to a greater amplitude of the response but not to a prolonged action. Hypophyseal desensitization to a subsequent stimulation was induced when the interval between stimulations was 30 min, but did not occur when lengthened to 60 or 120 min. Injection of a single dose of cLHRH-I or -II in vivo at doses of 10 and 0.1 nmol/kg body weight stimulated increases in the plasma concentration of LH and testosterone initiated within 1 or 10 min after injection respectively. As in vitro, cLHRH-II induced greater responses, which were dose-related, than did cLHRH-I. However, this difference could be attributed to a greater potency of cLHRH-II and to a more prolonged action. At the 10 nmol/kg dose, the shape of the LH response to cLHRH-II changed; it consisted of an initial increase during 10 min after injection, followed by a more sustained phase during which LH levels were still increasing between 20 and 60 min after injection. In contrast, after an injection of cLHRH-I at doses of 10 or 0.1 nmol/kg or cLHRH-II at a dose of 0.1 nmol/kg, LH levels were at a peak within 5 min and thereafter declined gradually.(ABSTRACT TRUNCATED AT 250 WORDS)

Comparison of mammalian luteinizing hormone releasing hormone (LH-RH), and of an analog (ICI 118630), on luteinizing hormone and ovarian steroid (progesterone, oestradiol) secretions in laying hens. (Gallus domesticus).

This experiment was conducted to compare the luteinizing hormone (LH), progesterone (P4) and oestradiol (E2) release in response to injections of various doses of synthetic mammalian luteinizing hormone-releasing hormone (LH-RH) and of an LH-RH agonist, ICI 118630, administered to laying hens 4 to 9 hours after a mid-sequence ovulation. Plasma LH increased significantly within 10 minutes of injection of either compound whereas any increases in plasma steroid concentrations were discerned later, at approximately minutes post-injection. No dose-response relationship was found for either compound with respect to LH release, but ICI 118630 appeared more potent than LH-RH. This analog also produced a greater mean incremental rise in plasma progesterone, but not oestradiol, than LH-RH, and this was found in animals injected at a time when the largest ovarian follicle was not mature. These result suggest that ICI 118630 is a more potent releasing hormone in the hen at the level of the pituitary, and that it may have a stimulating effect on ovarian progesterone secretion.

Respective effects of chicks and nest on behavior and hormonal concentrations of incubating domestic hens.

Removal of incubating hens from their nestboxes or substitution of chicks for eggs disrupt incubation; prolactin levels decrease whereas luteinizing hormone and gonadal steroid concentrations tend to rise. The present experiment was undertaken to determine the relative influence of removing the nest (nest-deprived hens), adding chicks (maternal hens), or both (maternal nest-deprived hens), on the behavior and hormone concentrations of incubating hens. The results confirm that nest removal, as well as adding chicks, stops incubation. No differences were found between maternal and maternal nest-deprived hens' behavioral responses or hormonal concentrations. These results do not support the hypothesis that the drop of plasma prolactin observed in maternal hens is the consequence of the nest abandonment. In contrast, nest-deprived hens presented lower concentrations of prolactin and higher concentrations of estradiol than maternal and maternal nest-deprived hens. Moreover, the nest-deprived hens presented lower concentrations of prolactin and higher concentrations of LH and estradiol than the hens given chicks. We assume that physical contact with chicks, during brooding bouts, slows down the decrease of prolactin secretion and inhibits LH and estradiol release at the end of incubation.

LH responses to chicken luteinizing hormone-releasing hormone I and II in laying, incubating, and out of lay turkey hens.

An experiment was conducted to assess the relative in vivo and in vitro activities of chicken LH-RH-I and -II in laying, incubating and out-of-lay turkey hens. The highest plasma concentrations of LH were measured in laying turkey hens, whereas hypophyseal concentrations were highest in incubating hens (I) and lowest in the laying hens at the end of the laying period (EL). Hypophyseal and plasma concentrations of LH decreased with aging in laying hens (L) and the greater decrease occurred in the hypophyses. An in vitro hypophyseal acute challenge with 2-min pulses of cLHRH I or II (10(-7) M) using a perifusion technique resulted in an increase in the release of LH in out-of-lay (OL) and incubating (I) hens, but not in laying (L) hens. Although both peptides elicited comparable responses in I hens, cLHRH II was more effective in OL hens. This difference was attributable to a greater amplitude of the response, whose duration was unchanged. Hypophyseal desensitization to a subsequent stimulation was observed in OL hens when the interval between stimulations was 30 min, but this did not occur at 60- or 120-min intervals. In vivo, the injection of cLHRH I or II, at doses of 10(-8) and 10(-10) M/kg B.W. stimulated increases in the plasma concentrations of LH, which were initiated within 1 min of injection in OL and I hens but from 5 to 20 min postinjection in L hens. The responses were dose-related and greater immediate responses were measured with cLHRH I than with cLHRH II. Also, after the injection of cLHRH II at the 10(-8) M/kg B.W. dose, the shape of the LH response consisted of an initial increase, followed by a more sustained phase during which LH concentrations were either stable (I hens) or continued to increase (L and OL hens) from 20 to 60 min after injection. In contrast, the injection of cLHRH I at doses of 10(-8) or 10(-10) M/kg or cLHRH II at a dose of 10(-10) M/kg in I and OL hens, produced a peak of LH concentrations in plasma within 5 min and thereafter declined gradually. The difference in the in vivo responses to LHRH I and II could not be attributed to a greater potency of cLHRH II, but to a more prolonged action. In summary, the responses to both forms of chicken LH-RH varies markedly with the stage of the reproductive cycle (L, I, and OL) and differs between the in vivo and in vitro situations. Although cLHRH II may be more active than cLHRH I, controversy still surrounds its precise physiological role.

Changes in endocrinological parameters and production performances in Turkey hens (Meleagris gallopavo) submitted to different broody management programs.

Occurrence of broodiness, egg production, feed consumption index and changes in the LH and prolactin levels were compared for turkey hens raised under 3 broody management programs. All hens (n=60 per group) were regularly ejected from the nest (basic treatment). Moreover, one group of hens was also rotated weekly (partial treatment), while in the third group (full treatment) hens showing broody symptoms were also identified daily and isolated for 24 hours. The percentage of hens which were identified as broody at least once was similar under the 3 programs (30%), although the use of the full and partial programs were more effective in inducing the disruption and/or preventing further expression of broody behavior. Plasma LH concentrations decreased progressively throughout the reproductive cycle under the 3 treatments. Maximum plasma concentrations of prolactin were measured between the 5th and the 10th weeks of production; higher concentrations of prolactin were maintained for a longer period under the full treatment, while a lower amplitude rise in plasma prolactin concentrations was observed in the hens submitted to the partial treatment. The egg production and feed consumption indices were lower and higher, respectively, under the full treatment than the basic treatment and partial treatments. We conclude that management programs need to be carefully evaluated under commercial conditions not only with respect to broodiness expression but also to egg production.

Prevention of incubation behavior expression in turkey hens by active immunization against prolactin.

The consequences of active immunization against prolactin on expression of incubation, reproductive performance and hormonal profiles were evaluated in turkey hens. Hens were injected weekly for 4 wk starting 8 wk before being submitted to a stimulatory photoperiod and 3 times thereafter at intervals of 4 to 5 wk. The hens were injected i.d. with 0.5 mL of a mixture diluted half in Freund's adjuvant. The mixture was prediluted in .9% saline and contained 100 micrograms of a fusion protein (GST-tPRL), GST, oPRL or vehicle. The results indicate that active immunizations with GST-tPRL or oPRL both induce production of specific prolactin antibodies. The onset of egg production was unaffected but higher egg production was observed for the GST-tPRL immunized hens. No GST-tPRL immunized hens expressed incubation behavior, whereas 20 to 30% of hens in the other experimental groups did so. Apparent hyperprolactinemia was detected by RIA for the GST-tPRL immunized groups starting before photostimulation and lasting until Week 10 of egg production but not in other groups. No significant differences were observed in either plasma LH or estradiol concentrations of immunized and nonimmunized turkey hens. In conclusion, both GST-tPRL or oPRL induced the production of antibodies against prolactin in turkey hens. However, only active immunization using GST-tPRL induced higher antibody titers as well as full prevention of incubation behavior expression. Such a pharmacological approach is of great practical interest, although its uses need to be carefully evaluated under commercial conditions.

Effects of an LH-RH agonist on reproductive responses and endocrinological parameters in landais ganders.

Semen quantitative (sperm production) and qualitative parameters (percentage of live and normal spermatozoa, sperm motility, egg fertility and hatchability), as well as hormonal parameters (LH and testosterone plasma concentrations) were compared for landais ganders, which were treated or not, with an LH-RH agonist prior to being sexually active. Treatment with the LH-RH agonist at this physiological stage delayed the onset of sperm production in some of the treated males. Although, comparable data were obtained during the first half of the reproductive period, treatment with the LH-RH agonist maintained sperm output at higher levels during its second half. Although the percentage of normal and live spermatozoa, sperm motility and true hatchability did not differ, the LH-RH agonist treatment had a positive effect on gosling production because of the higher fertility of the treated birds during the second part of the reproductive period. Treatment induced a large short-term decrease in testosterone levels followed by a rebound, leading to higher levels during the second half of the reproductive period. We conclude that treatment of ganders with an LH-RH agonist partially prevented the naturally occurring decline in sperm production and induced an increase in the rate of fertility rates during the second half of the productive period.

Responses of luteinizing hormone and prolactin plasma concentrations to oral administrations of clomiphene citrate in broody turkey hens.

Plasma levels in luteinizing hormone (LH) and prolactin were measured after oral administration of clomiphene-citrate (CC) at doses of 6 and 12 mg/kg BW/day for 5 consecutive days to broody turkey hens. No significant changes in prolactin levels were measured following treatment. The LH concentrations were significantly decreased (P less than .05) following administration of the 12-mg/kg dose. These results were interpreted as an indication of oestrogenic-like, rather than antioestrogenic-like, activity of CC when administrated to turkey hens at this particular physiological stage.

Effects of environmental and social factors on incubation behavior, endocrinological parameters, and production traits in turkey hens (Meleagris gallopavo).

Hens raised in three different environments were assessed for changes in egg production performance, the rate of incubation behavior expression, and plasma levels of luteinizing hormone (LH) and prolactin for 20 wk following the laying of the first egg. The environments were individual (IFP) or collective floor pens (CFP) and individual battery cages (Cp and Cnp). The hens from three experimental groups (IFP, CFP, and Cp) were transferred from a short (6 h) to a long (14 h) photoperiod, whereas the ones from the remnant (Cnp) were left under a short one. Increase of the photoperiod induced significant increases (P < 0.05) in levels of prolactin and LH after 1 d, and resulted in the onset of egg laying in a delay of 14 d in all groups. However, the overall egg laying performance was highest for the IFP hens. The CFP and IFP hens laid 98 and 24% of their eggs inside the nest boxes, respectively. The hens raised in battery cages did not express incubation behavior, whereas 50 and 33%, respectively, of the CFP and IFP hens did. During the 1st wk of egg laying, levels of prolactin increased for all photostimulated hens but to a greater extent for CFP hens. Higher increases in levels of prolactin were associated with the expression of incubation behavior; however, prolactin levels of nonincubating laying hens were also higher under the CFP treatment. It appears that the rate of expression of incubation behavior, as well as changes in the plasma levels of prolactin and LH throughout an egg production period, are dependent upon rearing conditions in turkey hens.

Follicular control of oviposition in the hen.

Ligation of the largest preovulatory follicle (F1) or the second largest preovulatory follicle (F2) delayed terminal oviposition for at least 2 h. Ligation of the fourth or fifth largest preovulatory follicle had no effect on the time of terminal oviposition. The conclusion reached was that some factor produced by the most mature follicles in the ovarian hierarchy influenced the timing of oviposition. Since the terminal oviposition occurs independently of ovulation, the authors concluded that the influence of the F1 and F2 follicles was independent of any preovulatory endocrine changes.

Variation of chicken technological meat quality in relation to genotype and preslaughter stress conditions.

The present study was aimed at estimating the genetic variability between lines of breast and thigh meat quality (pH decline, color, drip loss, and curing-cooking yield) by comparing a slow-growing French label-type line (SGL) and a fast-growing standard line (FGL) of chickens exposed to different preslaughter stress conditions. The birds were slaughtered under optimal conditions or after exposure to 2 h of transport or acute-heat stress (2 h at 35 degrees C). Relationships between meat quality and stress sensitivity were investigated by measuring struggle during shackling and tonic immobility (TI) duration, 1 wk before slaughter, as an indicator of the basal level of fear of the birds. Although most of the meat quality indicators varied between the 2 lines, differences were muscle dependent. In concordance with a lower ultimate pH, curing-cooking yield of thigh meat was decreased for the FGL birds. In contrast, these birds had a higher curing-cooking yield and a lower drip loss of breast meat resulting from a less rapid pH decline in this muscle compared with SGL birds. Thigh meat characteristics were influenced by both preslaughter stresses, but no significant effects were detected for breast meat. The main effect of heat stress in thigh meat was a decrease of the ultimate pH and led to paler color and lower curing-cooking yield; opposite effects were obtained for transport. Breast meat was much more sensitive to physical activity of birds on the shackle line. Longer durations of wing flapping on the shackle line gave more rapid initial pH decline. Whatever the line, no relationship between TI duration and meat quality characteristics or activity was observed. The present study suggested that SGL birds could be at disadvantage due to more struggle during shackling and accelerated postmortem glycolysis, which is detrimental to the quality of breast meat.

Detection of QTL controlling metabolism, meat quality, and liver quality traits of the overfed interspecific hybrid mule duck.

The mule duck, an interspecific hybrid obtained by crossing common duck (Anas platyrhynchos) females with Muscovy (Cairina moschata) drakes, is widely used for fatty liver production. The purpose of the present study was to detect and map single and pleiotropic QTL that segregate in the common duck species, and influence the expression of traits in their overfed mule duck offspring. To this end, we generated a common duck backcross (BC) population by crossing Kaiya and heavy Pekin experimental lines, which differ notably in regard to the BW and overfeeding ability of their mule progeny. The BC females were mated to Muscovy drakes and, on average, 4 male mule ducks hatched per BC female (1600 in total) and were measured for growth, metabolism during growth and the overfeeding period, overfeeding ability, and the quality of their breast meat and fatty liver. The phenotypic value of BC females was estimated for each trait by assigning to each female the mean value of the phenotypes of her offspring. Estimations allowed for variance, which depended on the number of male offspring per BC and the heritability of the trait considered. The genetic map used for QTL detection consisted of 91 microsatellite markers aggregated into 16 linkage groups (LG) covering a total of 778 cM. Twenty-two QTL were found to be significant at the 1% chromosome-wide threshold level using the single-trait detection option of the QTLMap software. Most of the QTL detected were related to the quality of breast meat and fatty liver: QTL for meat pH 20 min post mortem were mapped to LG4 (at the 1% genome-wide significance level), and QTL for meat lipid content and cooking losses were mapped to LG2a. The QTL related to fatty liver weight and liver protein and lipid content were for the most part detected on LG2c and LG9. Multitrait analysis highlighted the pleiotropic effects of QTL in these chromosome regions. Apart from the strong QTL for plasma triglyceride content at the end of the overfeeding period mapped to chromosome Z using single-trait analysis, all metabolic trait QTL were detected with the multitrait approach: the QTL mapped to LG14 and LG21 affected the plasma cholesterol and triglyceride contents, whereas the QTL mapped to LG2a seemed to impact glycemia and the basal plasma corticosterone content. A greater density genetic map will be needed to further fine map the QTL.

Minimising pain in farm animals: the 3S approach - 'Suppress, Substitute, Soothe'.

Recently, the French National Institute for Agricultural Research appointed an expert committee to review the issue of pain in food-producing farm animals. To minimise pain, the authors developed a '3S' approach accounting for 'Suppress, Substitute and Soothe' by analogy with the '3Rs' approach of 'Reduction, Refinement and Replacement' applied in the context of animal experimentation. Thus, when addressing the matter of pain, the following steps and solutions could be assessed, in the light of their feasibility (technical constraints, logistics and regulations), acceptability (societal and financial aspects) and availability. The first solution is to suppress any source of pain that brings no obvious advantage to the animals or the producers, as well as sources of pain for which potential benefits are largely exceeded by the negative effects. For instance, tail docking of cattle has recently been eliminated. Genetic selection on the basis of resistance criteria (as e.g. for lameness in cattle and poultry) or reduction of undesirable traits (e.g. boar taint in pigs) may also reduce painful conditions or procedures. The second solution is to substitute a technique causing pain by another less-painful method. For example, if dehorning cattle is unavoidable, it is preferable to perform it at a very young age, cauterising the horn bud. Animal management and constraint systems should be designed to reduce the risk for injury and bruising. Lastly, in situations where pain is known to be present, because of animal management procedures such as dehorning or castration, or because of pathology, for example lameness, systemic or local pharmacological treatments should be used to soothe pain. These treatments should take into account the duration of pain, which, in the case of some management procedures or diseases, may persist for longer periods. The administration of pain medication may require the intervention of veterinarians, but exemptions exist where breeders are allowed to use local anaesthesia (e.g. castration and dehorning in Switzerland). Extension of such exemptions, national or European legislation on pain management, or the introduction of animal welfare codes by retailers into their meat products may help further developments. In addition, veterinarians and farmers should be given the necessary tools and information to take into account animal pain in their management decisions.

Effects of housing conditions during the rearing and laying period on adrenal reactivity, immune response and heterophil to lymphocyte (H/L) ratios in laying hens.

This study was conducted to evaluate the effect of early rearing conditions on physiological, haematological and immunological responses relevant to adaptation and long-term stress in white Leghorn hens with intact beaks housed in furnished cages (FC) or conventional cages (CC) during the laying period. Pullets were cage reared (CR) or litter floor reared (FR). From 16 to 76 weeks of age, hens were housed in FC (eight hens per cage) or in CC (three hens per cage). As measures of long-term stress at the end of the laying period, adrenal reactivity was quantified by assessing corticosterone responses to adrenocorticotropin challenge, and immune response was assessed by measuring antibody responses after immunization with sheep red blood cells (SRBC) and keyhole limpet haemocyanin (KLH). Heterophil to lymphocyte (H/L) ratio was employed as an indicator of stress. Rearing conditions significantly affected anti-SRBC titres (P < 0.0001) and tended to affect H/L ratios (P = 0.07), with the highest values found in FR hens. Layer housing affected H/L ratio (P < 0.01); the highest ratio was found in FR birds housed in FC during the laying period. This study shows that early rearing environment affects immunological indicators that are widely used to assess stress in laying hens. However, while results on H/L ratio indicated that FR birds experienced more stress particularly when they were housed in FC during the laying period, the immune responses to SRBC in FR hens was improved, indicating the opposite. This contradiction suggests that the effects on immune response may have been associated with pathogenic load due to environmental complexity in FR and FC hens rather than stress due to rearing system or housing system per se.

Intensity and duration of corticosterone response to stressful situations in Japanese quail divergently selected for tonic immobility.

Two genotypes of Japanese quail have been divergently selected since the 1980s for long (LTI) or short (STI) duration of tonic immobility [Mills, A.D., Faure, J.M., 1991. Divergent selection for duration of tonic immobility and social reinstatement behavior in Japanese quail (Coturnix coturnix japonica) chicks. J. Comp. Psychol. 105(1), 25-38.], an unlearnt catatonic state characteristic of a behavioral fear response ([Jones, R.B., 1986. The tonic immobility reaction of the domestic fowl: a review. World's Poult. Sci. J. 42(1), 82-97.]; [Mills, A.D., Faure, J.M., 1991. Divergent selection for duration of tonic immobility and social reinstatement behavior in Japanese quail (Coturnix coturnix japonica) chicks. J. Comp. Psychol. 105(1), 25-38.]). The results of several behavioral tests conducted in LTI and STI quail have led to the conclusion that LTI quail are more fearful than STI quail [Faure, J.M., Mills, A.D., 1998. Improving the adaptability of animals by selection. In: Grandin, T. (Eds.), Genetics and the behavior of domestic animals. Academic Press, San Diego, pp. 235-264.]). However, few studies to date have focused on the Hypothalamic-Pituitary-Adrenal (HPA) axis response to stressful situations in LTI and STI quail, although the HPA axis is involved in fear responses [Siegel, H.S., 1971. Adrenals, Stress and the Environment. World's Poult. Sci. J. 27, 327-349.]. The corticosterone (CORT) response to various putatively stressful situations was therefore assessed in LTI and STI genotypes of quail in order to investigate their HPA axis reactivity to stress. Repeated induction of TI or 1 min manual restraint induced significant and comparable increases in CORT levels in both genotypes as a TI response. On the other hand, higher CORT responses were found in STI than in LTI quail when the manual restraint period lasted for 2 min or after restraint in a crush cage. Maximum CORT responses and genotype differences were maintained throughout the latter test even when it lasted for 120 min. Investigation of the CORT response to a single TI episode showed that CORT levels at the end of TI were negatively correlated with TI duration. Other experimental contexts consisting of isolation in a familiar or novel environment or the presentation of a novel object induced slight but significant and comparable increases in CORT response in both genotypes, whereas change of cagemates did not. In conclusion, the present findings indicate that differences in HPA axis response are observed between LTI and STI genotypes when quail are submitted to intense stressors, resulting in a high and prolonged CORT response. By contrast, plasma corticosterone concentrations do not differ between STI and LTI quail in response to stressful situations of lower intensity, which evoke responses limited in amplitude and duration. Genetic selection for divergent duration of TI has thus affected the HPA axis response to stress, and LTI and STI quail constitute an interesting model to investigate genetic variability of HPA axis activity in birds. More specifically, these genotypes of quail could be used to investigate the occurrence of functional differences at different levels of the HPA axis in order to explain the present findings.