R. L. Dennis

Serotonergic Mediation of Aggression in High and Low Aggressive Chicken Strains

Serotonin (5-HT) regulates aggressive behavior via binding to its receptors, such as 5-HT1A and 1B, in humans and rodents. Here we investigate the heritable components of 5-HT regulation of aggressiveness in chickens, utilizing 3 distinct genetic strains. In this study, we used 2 divergently selected strains (high and low group productivity and survivability, respectively; HGPS and LGPS) and a third strain, Dekalb XL (DXL), an aggressive out-group. Hens were paired within the same strain. At 24 wk of age, the subordinate of each pair received a daily i.p. injection of NAN-190 (0.5 mg/kg, a 5-HT1A antagonist), GR-127935 (0.5 mg/kg, a 5-HT1B antagonist), or saline (control) for 5 consecutive days. Frequency of aggressive behaviors was increased in the hens of DXL and LGPS treated with 5-HT1A antagonist and in the HGPS hens treated with 5-HT1B antagonist. The 5-HT1B antagonist-treated HGPS hens and 5-HT1A antagonist-treated LGPS hens also displayed increased feather pecking, but neither antagonist had an effect on feather pecking of DXL hens. This may suggest that multiple mediating factors alter feather pecking behaviors. Among the controls, LGPS hens have higher epinephrine levels than HGPS or DXL hens, indicative of the inferior stress-coping ability of LGPS hens. Treatment with 5-HT1B antagonist reduced epinephrine in LGPS hens but not in DXL or HGPS hens, suggesting a role of 5-HT1B in stress regulation in LGPS hens. The results provide evidence for different heritable serotonergic mediation of aggressive behaviors and stress coping in chickens.

Infrared beak treatment method compared with conventional hot-blade trimming in laying hens

Infrared lasers have been widely used for noninvasive surgical applications in human medicine, and their results are reliable, predictable, and reproducible. Infrared lasers have recently been designed for the express purpose of providing a less painful, more precise beak-trimming method compared with conventional beak trimming. This study was designed to examine the potential of the infrared (IR) beak treatment to provide a welfare-friendly alternative to the conventional hot-blade method for chickens. The birds were beak trimmed by IR at the hatchery or by hot blade at 7 to 10 d of age in a commercial production setting, in accordance with standard procedures. The beak morphology and associated physiological characteristics, including production and aggressive behavior of the birds, were analyzed at 30 wk of age. There was no difference in egg production or bird BW between the 2 beak-trimming treatments. Birds also exhibited no difference in stress physiology measured in the study, such as fluctuating asymmetry and heterophil and lymphocyte profiles. However, birds receiving the IR treatment showed a superior feather condition and reduced aggressiveness under high light intensity, even though they had longer beak stumps. The results may indicate that the IR beak treatment may reduce the damage done by aggressive pecking and feather pecking. Indeed, IR trimming may provide a more welfare-friendly alternative to conventional beak trimming without compromising productivity.

Appearance Matters: Artificial Marking Alters Aggression and Stress

Artificial marking of animals for identification is frequently employed by researchers in the behavioral, biomedical, agricultural, and environmental sciences. The impact of artificial marking on experimental results is rarely explicitly considered despite evidence demonstrating that changes in phenotypic appearance can modify animal behavior and reproductive success. Here we present evidence that artificial marking of individuals within a social group has frequency-dependent effects on the behavior and physiology of domestic fowl (Gallus gallus domesticus). We demonstrate that when only 20 or 50% of individuals within a group were artificially marked, the marked birds received more aggression and had lesser body mass than the unmarked individuals within the same group. Furthermore, in groups in which only a small proportion of the individuals were marked, we report altered plasma epinephrine and dopamine levels in marked individuals. These effects of marking were imperceptible when all birds in a group were marked. This finding has important implications for animal research because, when only a subset of group members is artificially marked and used for data collection, the results obtained may not be representative of the population.

Effects of raclopride on aggression and stress in diversely selected chicken lines.

Genetic selection for chickens of high (HGPS) and low (LGPS) group productivity and survivability, resulted in two distinct genetic lines characterized by differences in cannibalism, flightiness, and immunocompetence. Additionally, birds exhibited differences in behaviour and social stress coping strategy. HGPS birds have a superior stress coping strategy compared with birds of LGPS or Dekalb XL (DXL), a commercial strain. Line differences in stress response and behaviour could be due to selection-induced differences in expression of the dopaminergic system. The dopamine (D2) receptor, an integral part of the dopaminergic system, was hypothesized to be a key contributory factor of the stress response. We tested this hypothesis by injecting either a D2 antagonist (raclopride) or saline in the dominant individual in pair-housed birds for 10 days and examining stress coping ability. Results showed that dominant birds of all strains showed a reduced frequency of aggressive pecks on subordinates following raclopride injection. In contrast, subordinates paired with raclopride-injected birds increased pecking frequency. Two days after stopping injections, LGPS and DXL birds returned to pre-injection levels of aggressive threats, while HGPS birds maintained depressed frequency of threats. Strain differences in aggressive responsiveness coincided with increased epinephrine levels in raclopride treated LGPS birds relative to control LGPS birds, but not by HGPS and DXL birds. Our findings suggest a functional linkage between the genetic basis of stress coping ability and the dopamine regulation of aggressive responsiveness. The data further indicate that the sympathetic-adreno-medullary axis is directly involved in regulating both stress coping strategy and aggressiveness.

Alterations to Embryonic Serotonin Change Aggression and Fearfulness

Prenatal stress can alter the serotonin (5-HT) system in the developing and adult brain and lead to mood and behavioral disorders in children and adults. The chicken provides a unique animal model to study the effects of embryonic stressors on childhood and adolescent behavior. Manipulations to the egg can be made in the absence of confounding maternal effects from treatment. Eggs were injected with 50 μL of excess 5-HT (10 μg/egg), 8-OH-DPAT (a 5-HT1A receptor agonist; 16 μg/egg), or saline on day 0 prior to the 21 days incubation. Injections were performed at 0.5 cm below the shell. Behavior was analyzed at 9 weeks of age and again at the onset of sexual maturity (18 weeks). Hens treated with excess embryonic 5-HT exhibited significantly less aggressive behaviors at 9 weeks of age compared to both 5-HT1A agonist treated and saline hens (P < 0.05), and at 18 weeks of age compared to saline control birds only (P < 0.05). Excess embryonic 5-HT also increased fearfulness response (P < 0.05) as tested by duration of tonic immobility. Increased degree of fluctuating asymmetry at 18 weeks in 5-HT-treated birds (P < 0.05) suggests that excess 5-HT in early embryonic stages may create a developmental instability, causing phenotypic variations. These results showed that modification of the serotonergic system during early embryonic development alters its functions in mediating aggressive and fearful or anxious behaviors. Prenatal modification of the serotonergic system has long lived implications on both physiology and behavior, especially aggressive and fearful behaviors.

The dopaminergic system and aggression in laying hens

The dopaminergic system is involved in the regulation of aggression in many species, especially via dopamine (DA) D1 and D2 receptor pathways. To investigate heritable differences in this regulation, 2 high aggressive strains [Dekalb XL (DXL) and low group egg productivity and survivability (LGPS)] and one low aggressive strain (low group egg productivity and survivability; HGPS) of laying hens were used in the study. The HGPS and LGPS lines were diversely selected using group selection for high and low group production and survivability. The DXL line is a commercial line selected through individual selection based on egg production. Heritable differences in aggressive propensity between the strains have been previously assessed. The birds were pair housed within the same strain and labeled as dominant or subordinate based on behavioral observation. For both experiments 1 and 2, behavioral analysis was performed on all 3 strains whereas neurotransmitter analysis was performed only on the most aggressive (DXL) and least aggressive (HGPS) strains. In experiment 1, the subordinate birds were treated with D1 agonist, D2 agonist, or saline controls (n = 12). In experiment 2, the dominant birds from a separate flock were treated with D1 antagonist, D2 antagonist, or saline controls (n = 12). Treatment-associated changes in aggressive behaviors and central neurotransmitters were measured. Aggression was increased in all strains in response to D1 agonism but increased only in the less aggressive HGPS birds with D2 agonism. Aggression was decreased and hypothalamic serotonin and epinephrine were increased in birds from all strains treated with D2 receptor antagonist. The D1 receptor antagonism elicited different behavioral and neurotransmitter responses based on the aggressive phenotype of the genetic strains. Aggressive strains DXL and LGPS but not the HGPS strain decreased aggressiveness following antagonism of the D1 receptor. The data show evidence for distinct neurotransmitter regulation of aggression in high and low aggressive strains of hens through different receptor systems. These chicken lines could provide new animal models for the biomedical investigation of the genetic basis of aggression.

Effects of different infrared beak treatment protocols on chicken welfare and physiology

Infrared beak trimming provides an alternative to conventional trimming, purporting to provide a welfare-friendly means of trimming. The infrared system can be adjusted to use multiple plate and power settings. In the present study, we used 2 different plate sizes (27/23C, less severe; 25/23C, more severe) with each of 3 power settings: high (52), moderate (48), and low (44). These birds, along with conventionally (hot blade; HB) trimmed birds were maintained in an industry egg-laying facility. Physiological and behavioral measures were taken at 5, 10, 20, and 30 wk. All birds followed a similar growth curve; birds from the 27/23C (48) protocol were the heaviest across all ages and 25/23C (44) birds were the lightest. Upper and lower beak growth curves showed birds trimmed with 25/23C protocols had shorter upper and lower beaks compared with 27/23C protocols or HB. Birds trimmed using 27/23 (44) and (48) had consistently longer upper and lower mandibles. Amount of feed wasted was greatest in HB and 27/23C birds and tended to be reduced in 27/23 (48) and 25/23 (48) and (52) birds (P < 0.10). Beak-related behaviors (eating, drinking, and pecking) were measured to observe the effects of trimming protocol on beak usage, which could indicate beak pain or morphological changes that inhibit normal behaviors. Walking behavior was also measured to assess overall activity. Behavior analysis revealed that compared with HB-trimmed birds, those of 27/23C protocols walked and drank more at a young age. At 5 and 10 wk of age, a test feather was attached to the cage and pecking at as well as the damage score of the feather were determined. Birds from 27/23C (44) and (48) protocols pecked significantly more at the feather than HB, whereas HB and 25/23C (52) birds had the highest damage score. The results from the study suggest that infrared protocols can be optimized for superior productivity as well as animal well-being.

Effect of perches on liver health of hens

Fatty liver is a common energy metabolic disorder in caged laying hens. Considering that the egg industry is shifting from conventional cages to alternative housing systems such as enriched cages, the objective of this study was to determine the effects of perches on fat deposition and liver health in laying hens. Three hundred twenty-four 17-wk-old White Leghorn hens were housed in 1 of 4 treatments with 9 hens per cage. Treatment 1 hens never had access to perches during their life cycle. Treatment 2 hens had access to perches during the pullet phase only. Treatment 3 hens had access to perches during the laying phase only. Treatment 4 hens always had access to perches. Liver weight, abdominal fat pad weight, BW, liver fat, and circulating alanine transaminase, aspartate transaminase, and adiponectin were determined. Provision of perches during either the rearing or laying phase did not affect liver health in 71-wk-old hens. However, perch access compared with no perch access during the egg laying phase reduced relative fat pad weight. These results suggest that providing perches as a means of stimulating activity reduced abdominal fat deposition in caged hens during the laying period. However, perch access in caged hens was ineffective in reducing fat deposition in the liver and altering enzyme activities related to improved liver function.

Effects of selective serotonin antagonism on central neurotransmission

Aggression and cannibalism in laying hens can differ in intensity and degree due to many factors, including genetics. Previous behavioral analysis of 2 strains of White Leghorns, DeKalb XL (DXL) and HGPS (a group-selected line for high group productivity and survivability), revealed high and low aggressive phenotypes, respectively. However, the exact genetic mechanisms mediating aggressiveness are currently unknown. Analysis of serotonin (5-HT) mediation of aggression in subordinate hens of these strains revealed increases in aggression in DXL hens following antagonism of the 5-HT1A receptor and in HGPS hens following antagonism of the 5-HT1B receptor. Here, we investigate the different neurotransmitter response in the hypothalamus and raphe nucleus mediating these aggressive responses to receptor antagonism. Elevated aggressive response to 5-HT1B antagonism by HGPS hens was also accompanied by a decrease in raphe nucleus dopamine (DA) and an increase in DA turnover. Increased aggressiveness in DXL hens did not coincide with a reduction in raphe nucleus 5-HT or turnover (as indicated by 5-hydroxyindoleacetic acid levels) following 5-HT1A antagonism. A reduction in 5-hydroxyindoleacetic acid (but not 5-HT) was seen in HGPS hens treated with 5-HT1A antagonist; however, these hens exhibited no change in aggressive behaviors. Our data show evidence of different heritable mechanisms of neurotransmitter regulation of aggressive response, specifically heritable differences in the interaction between 5-HT and catecholamines in regulating aggression.

Serotonin’s role in piglet mortality and thriftiness

Improving piglet survivability rates is of high priority for swine production as well as for piglet well-being. Dysfunction in the serotonin (5-HT) system has been associated with growth deficiencies, infant mortalities, or failure to thrive in human infants. The aim of this research was to determine if a relationship exists between infant mortality and failure to thrive (or unthriftiness), and umbilical 5-HT concentration in piglets. Umbilical blood was collected from a total of 60 piglets from 15 litters for analysis of 5-HT and tryptophan (Trp; the AA precursor to 5-HT) concentrations. Behavior was scan sampled for the first 2 days after birth. Brain samples were also taken at 8 h after birth from healthy and unthrifty piglets (n = 4/group). The raphe nucleus was dissected out and analyzed for 5-HT and dopamine concentrations as well as their major metabolites 5-hydroxyindoleacetic acid (5-HIAA) and homovanillic acid (HVA), respectively. Data were analyzed by ANOVA. Piglets that died within 48 h of birth (n = 14) had significantly lower umbilical blood 5-HT concentrations at the time of their birth compared to their healthy counterparts (n = 46, P = 0.003). However, no difference in Trp was detected (P 0.38). Time spent under the heat lamp and sleeping were positively correlated with umbilical 5-HT levels (P = 0.004 and P = 0.02, respectively), while inactivity had a negative correlation with 5-HT levels (P = 0.04). In the raphe nucleus, the center for brain 5-HT biosynthesis, unthrifty piglets had a greater concentration of 5-HIAA (P = 0.02) and a trend for higher concentrations of 5-HT (P = 0.07) compared with healthy piglets. Dopamine levels did not differ between thrifty and unthrifty piglets (P = 0.45); however, its metabolite HVA tended to be greater in unthrifty piglets (P = 0.05). Our results show evidence of serotonergic dysfunction, at both the central and peripheral levels, accompanying early piglet mortalities. These data suggest a possible route for intervention, via the 5-HT system, to improve piglet survivability. However, further research is required to validate this hypothesis.