Brain gain-Is the cognitive performance of domestic hens affected by a functional polymorphism in the serotonin transporter gene?

The serotonin transporter (5-HTT) plays an important role in regulating serotonergic transmission via removal of serotonin (5-HT) from synaptic clefts. Alterations in 5-HTT expression and subsequent 5-HT transmission have been found to be associated with changes in behaviour, such as fearfulness or activity, in humans and other vertebrates. In humans, alterations in 5-HTT expression have been suggested to be able to lead to better learning performance, with more fearful persons being better at learning. Similar effects of the variation in the 5-HTT on fearfulness have been found in chickens, and in this study, we investigated effects on learning. Therefore, we tested 52 adult laying hens, differing in their functional 5-HTT genotype (W/W, W/D and D/D) in an operant learning paradigm in three different phases (initial learning, reversal learning and extinction) and in a tonic immobility test for fearfulness. We found that the 5-HTT polymorphism affects the initial learning performance of laying hens, with homogeneous wild-type (W/W) hens being the slowest learners, and the most fearful birds. W/W hens, showed significantly more choices to solve the initial learning task (LME, p = 0.031) and had the highest latencies in a tonic immobility test (p = 0.039), indicating the highest fearfulness. Our results provide interesting first insights into the role of 5-HTT in chickens and its sensitive interaction with the environment. We further suggest that the 5-HTT gene can be an interesting target gene for future breeding strategies as well as for further experimental studies.

Productive performance, perching behavior, keel bone and other health aspects in dual-purpose compared to conventional laying hens.

Several alternatives to avoid killing male day-old chicks are available. One of these alternatives is to keep dual-purpose chicken strains. The aim of this study was to compare dual-purpose hens (Lohmann Dual, LD) with conventional laying hens (Lohmann Tradition, LT) in terms of performance, animal welfare parameters such as keel bone state and foot pad dermatitis, and perching behavior. We expected a generally equal or even better performance of the dual-purpose hens except for laying performance. Four hundred female day-old chicks were housed in 6 pens (3 pens per strain) and reared until 54 wk of age. Each pen offered a littered area, elevated slatted manure pit, elevated wooden frame with perches or grids and nest boxes on the manure pit. The wooden frame was alternately equipped with perches or grids. The elevated manure pit as well as the elevated structure was accessible via ramp. Productive performance parameters like mortality, total number of eggs and body weight were assessed periodically. In week 49, 132 hens (66 hens per strain) were randomly selected for radiography of the keel bone and assessment of plumage and foot pad state. Perching behavior was analyzed via scan sampling during rearing and laying period, respectively. Statistical analyzes were done with Linear Mixed Effect Model and General Linear Mixed Model. LD had a higher radiographic density than LT hens (P = 0.0016), other keel bone parameters (fracture score, P = 0.36; deformation, P = 0.83) showed no differences. The vast majority of fractures occurring in both strains were located in the caudal part of the keel bone. During the laying period, usage of elevated structures was higher with grids compared to perches (P < 0.001) and in LD compared to LT (P = 0.01). Some animal welfare problems were less frequent in LD compared to LT hens while other problems did not differ between the 2 strains or were even more frequent in LD hens. Grids may be more suitable as resting area than perches and may possibly help to decrease the prevalence of keel bone damage.

Estradiol-17ß Is Influenced by Age, Housing System, and Laying Performance in Genetically Divergent Laying Hens (Gallus gallus f.d.).

The estrogen estradiol-17ß is known as one of the major gonadal steroid hormones with different functions in reproduction. In this study we analyzed estradiol-17ß concentration in laying hens of four pure bred chicken laying lines at four different time intervals of the laying period (17th-19th week of age, 33rd-35th week of age, 49th-51st week of age, and 72nd week of age). The high performing white egg (WLA) and brown egg (BLA) layer lines as well as the low performing white (R11) and brown (L68) layer lines were kept in both single cages and a floor housing system. We investigated whether there were differences in estradiol -17ß concentrations between lines at different ages that could be related to selection for high egg production or phylogenetic origin of the animals, and whether there was an influence of housing conditions on estradiol-17ß. Estradiol-17ß concentrations differed between high and low performing layer lines at all time intervals studied. High performing hens showed higher estradiol-17ß concentrations compared to low performing hens. In all lines, highest estradiol-17ß concentration was measured at their 49th to their 51st week of age, whereas the peak of laying intensity was observed at their 33rd to their 35th week of age. Additionally, hens with fewer opportunities for activity housed in cages showed higher estradiol-17ß concentrations than hens kept in a floor housing system with more movement possibilities. We could show that laying performance is strongly linked with estradiol -17ß concentration. This concentration changes during laying period and is also influenced by the housing system.

Learned control of urinary reflexes in cattle to help reduce greenhouse gas emissions.

Indiscriminate voiding of excreta by cattle contributes to greenhouse gas (GHG) emissions and soil and water contamination1,2. Emissions are higher in animal-friendly husbandry offering cattle more space2 - a trade-off we call the 'climate killer conundrum'. Voiding in a specific location (latrine) would help resolve this dilemma by allowing ready capture and treatment of excreta under more spacious farming conditions. For urination, toileting requires self-control and coordination of a complex chain of behaviors including awareness of bladder fullness, overriding of excretory reflexes, selection of a latrine and intentional relaxation of the external urethral sphincter3. Attempts to train toileting in cattle have so far been only partly successful4-6, even though their excretion and associated neurophysiological control are similar to those in species capable of toileting3. Similarly, very young infants have been considered incapable of self-initiated voiding, but they can be taught with extensive training7. Using a backward chaining, reward-based training procedure, we here show that cattle can control their micturition reflex and use a latrine for urination. Such self-control provides evidence that animals can learn to respond to and reveal internal experiences via appropriately trained operant behaviors, thereby providing another way to explore their subjective states.

Effects of an Elevated Platform on Welfare Aspects in Male Conventional Broilers and Dual-Purpose Chickens.

To avoid killing day-old male chicks, one possibility is to keep dual-purpose chicken strains. Here, the hens were kept for egg production, and the roosters were kept for meat production. Both sexes had moderate performances compared to the respective hybrid chicken strains. However, until now, little has been known about whether male dual-purpose chickens may profit from enrichment in the environment in which broiler chickens are raised under conventional conditions. This study aims to further investigate the suitability of elevated structures for dual-purpose chickens (Lohmann Dual) with moderate growth and for fast-growing male broiler chickens (Ross 308). In two consecutive trials, we kept 686 Ross and 672 Dual chickens in 24 compartments (2 trials × 2 strains × 6 compartments). Half of the compartments were equipped with elevated grid platforms at a height of 50 cm (enriched group). In the other half of the compartments, no platforms were installed (control group). We analyzed the usage of the elevated platforms by scan sampling and assessed animal-based (walking ability, plumage cleanliness, and foot health) and management-based (litter quality) indicators. Both strains showed increasing use of the elevated platforms from the first week of life onwards. However, the fast-growing chickens used the elevated platform less than the slow-growing chickens. At the end of the fattening period, the birds used the elevated grids more at night than during the daytime. Slow-growing chickens kept in enriched compartments showed a better walking ability. In general, slow-growing chickens had better plumage conditions and foot health compared to fast-growing chickens. Our results show that natural behaviors such as perching can be supported by offering elevated platforms and that animal-based indicators such as walking ability can be improved, at least in slow-growing chickens. Moreover, the use of an alternative chicken strain avoids killing day-old male chicks, and in addition, these chickens show fewer animal welfare problems than a conventional fattening strain. Thus, the use of male chickens of a dual-purpose strain can substantially contribute to improving animal welfare in broiler meat production.

Tasty straw pellets - Exploration of flavoured rooting material by pigs.

Tail biting is a common problem in pigs kept in conventional fully slatted pens. Suitable enrichment materials can help to prevent the occurrence of this behavioural disorder by encouraging pigs to increase exploration behaviour. We investigated whether additional flavours can increase exploration behaviour in undocked pigs. Therefore, we offered straw pellets flavoured with either fried onion (FO), strawberry (SB), ginger, almond (AL), vanilla or without flavour (control) during rearing (eight groups in total) and fattening (16 groups in total). Flavoured pellets were offered in an altering order during intervals of 1 week in material dispensers. Exploration duration at the material dispensers was continuously recorded via an ultra-high-frequency radio-frequency identification system. Pigs were weighed weekly and their tail lengths and tail injuries were scored in four categories. For analysis, changes in tail length scores compared to the previous week were calculated as Δ-tail length. The different flavours affected pigs' exploration durations in both rearing (factor flavour, P < 0.0001) and fattening (factor flavour, P < 0.0001). Highest exploration durations during rearing were recorded when straw pellets were flavoured with FO and AL compared to all others. During fattening, exploration duration was highest towards controls without significant difference to SB. Exploration durations additionally were affected by temporal effects, i.e. week and day during rearing (both factors: P < 0.0001) and fattening (both factors: P < 0.0001). During rearing, highest exploration durations were recorded in the first week and on the first day within week after changing the flavour of the straw pellets. During fattening, exploration durations varied between weeks and within weeks. The highest durations were recorded at the end of weeks, i.e. on the fifth and seventh days after material change. During rearing, tail injuries were affected by week (P < 0.0001). From the fourth week of rearing, the prevalence of tail injuries significantly increased. During the fattening period, tail injuries were affected by flavour (P < 0.05). Fattening pigs had fewest tail injuries after straw pellets flavoured with AL were offered. Altogether, based on the exploration durations, rearing pigs showed different preferences for the flavoured straw pellets but highly explored flavours differed between rearing and fattening pigs. Despite a weekly change of the flavour of straw pellets, exploration durations decreased during rearing but increased again in the transition between rearing and fattening. Thus, straw pellets with alternating flavours are a suitable possibility to provide environmental enrichment to pigs but will not prevent tail biting.

How can Cattle be Toilet Trained? Incorporating Reflexive Behaviours into a Behavioural Chain.

Untrained cattle do not defecate or urinate in defined locations. The toilet training of cattle would allow urine and faeces to be separated and stored, reducing climate-damaging emissions and improving animal health. In a proof-of-concept study, we evaluated a novel protocol for toilet training in cattle. Five heifer calves (and yoked controls) were trained in the voluntary (operant) behaviours of a toileting chain. Then, reflexive urinating responses were incorporated into the chain, with toileting signalled by a tactile (vibratory) stimulus. On 95% of occasions, the calves inhibited/interrupted urination when receiving the stimulus, and on 65% of these occasions, reinitiated urination in the latrine. Furthermore, during 63% of urinations in the latrine, the calves oriented to the reward location before any food was delivered, providing additional evidence that calves can be successfully toilet trained with food rewards. Yoked controls failed to learn most of the operant elements and all the reflexive responses of toilet training. The results show that reflexive behaviours can be incorporated into voluntary toileting sequences with cattle and extend the range of species that can be toilet trained. Future refinement of the protocol to allow training under practical farm conditions offers the potential to mitigate climate damage and improve animal health.

Conditionability of 'voluntary' and 'reflexive-like' behaviors, with special reference to elimination behavior in cattle.

Typically, cattle urinate and defecate with little or no control over time and place. The resulting excreta contributes to a range of adverse effects on the environment and the animals themselves. These adverse effects could be substantially ameliorated if livestock could be toilet trained. Toilet training requires an animal to suppress impending voiding (a reflexive-like behavior), move to a latrine (voluntary behavior) and reinitiate voiding. Here, we review the neurophysiological processes and learning mechanisms regulating toileting. The suppression and initiation of voiding occur primarily via the coordinated activity of smooth and striated anal and urinary sphincter muscles. The autonomic and somatic nervous systems, along with central processes, regulate these muscles. In several mammalian species, voluntary control of the sphincters has been demonstrated using classical and/or operant conditioning. In this review, we demonstrate that the neurophysiological and behavioral regulation of voiding in cattle is likely to be similarly conditionable. The management of excreta deposition in cattle could have major benefits for reducing livestock greenhouse gas emissions and improving animal health/welfare.

The Role of Egg Production in the Etiology of Keel Bone Damage in Laying Hens.

Keel bone fractures and deviations belong to the most severe animal welfare problems in laying hens and are influenced by several factors such as husbandry system and genetic background. It is likely that egg production also influences keel bone health due to the high demand of calcium for the eggshell, which is, in part, taken from the skeleton. The high estrogen plasma concentration, which is linked to the high laying performance, may also affect the keel bone as sexual steroids have been shown to influence bone health. The aim of this study was to investigate the relationship between egg production, genetically determined high laying performance, estradiol-17ß concentration, and keel bone characteristics. Two hundred hens of two layer lines differing in laying performance (WLA: high performing; G11: low performing) were divided into four treatment groups: Group S received an implant containing a GnRH agonist that suppressed egg production, group E received an implant containing the sexual steroid estradiol-17ß, group SE received both implants, and group C were kept as control hens. Between the 12th and the 62nd weeks of age, the keel bone of all hens was radiographed and estradiol-17ß plasma concentration was assessed at regular intervals. Non-egg laying hens showed a lower risk of keel bone fracture and a higher radiographic density compared to egg laying hens. Exogenous estradiol-17ß was associated with a moderately higher risk of fracture within egg laying but with a lower risk of fracture and a higher radiographic density within non-egg laying hens. The high performing layer line WLA showed a significantly higher fracture risk but also a higher radiographic density compared to the low performing layer line G11. In contrast, neither the risk nor the severity of deviations were unambiguously influenced by egg production or layer line. We assume that within a layer line, there is a strong association between egg production and keel bone fractures, and, possibly, bone mineral density, but not between egg production and deviations. Moreover, our results confirm that genetic background influences fracture prevalence and indicate that the selection for high laying performance may negatively influence keel bone health.

Comparison of cortisol concentrations in plasma and saliva in dairy cattle following ACTH stimulation.

The experiments described in this research communication compared cortisol concentrations in plasma and saliva samples collected from dairy cattle before and after an adrenocorticotropic hormone (ACTH) application. For that purpose, blood and saliva samples were collected from five dairy cows at determined time intervals before (490 min and down to 0 min) and after (10 min and up to 500 min) an ACTH application. Mean baseline cortisol concentrations were greater in plasma compared to saliva. The relative increases and decreases in plasma and salivary cortisol concentrations following ACTH were similar. After ACTH, we observed an increase in cortisol concentrations in plasma after 10 min and in saliva after 20 min. The time of peak concentrations after ACTH were reached at 70 and 80 min for plasma and saliva, respectively. After peak concentrations, values steadily declined and returned to baseline values at 169 ± 15 min in plasma and 170 ± 14 min in saliva. Ratios between salivary and plasma cortisol concentrations were on average 0.09 and did not change substantially during the ACTH challenge. There was a strong positive relationship between salivary and plasma cortisol concentrations. These results indicate that salivary cortisol concentrations can be a good indicator of ACTH-induced plasma cortisol concentrations in dairy cattle.

Behavioural consequences of divergent selection on general locomotor activity in chickens.

General locomotor activity is a highly variable phenotypic trait of animals. In domestic chickens it is different within and between breeds. The general locomotor activity is a substantially heritable trait and has been shown to be correlated with several other behavioural traits, such as for example feather pecking and anxiety in chickens. However, whether there is a relation between different levels of general locomotor activity and behavioural changes remained unclear. Therefore, a selection line model system has been established, where hens from the same founder population were selected over eight generations for either high or low general locomotor activity. The selection led to significant increases, respectively decreases in general locomotor activity and differences in growth. We here tested 128 hens of the 8th generation in three behavioural tests. We assumed fearfulness to be affected from selection on general locomotor activity, which we tested in a tonic immobility test. Socio-positive and socio-negative behaviours were tested in respective test paradigms. Fearfulness was higher in hens selected for high general locomotor activity. Social behavioural traits and feather pecking were not affected by selection for general locomotor activity. Evolutionary mechanisms that link fear and general locomotor activity are discussed and also why social behaviours and feather pecking seems not to be affected from selection on general locomotor activity. Our results provide interesting new insights on how selection on one trait, general locomotor activity, affects the behavioural phenotype in other dimensions too.

Effects of Elevated Grids on Growing Male Chickens Differing in Growth Performance.

Pullets, i. e., chickens of layer lines are often raised in housings equipped with perches. In contrast, broiler chickens most often are raised in a barren environment that lacks any three-dimensional structures, even though broilers also are motivated to use elevated structures. In addition, environmental enrichment may improve welfare problems in broiler chickens, such as skeletal disorders or contact dermatitis. Due to ethical reasons, currently there are attempts to fatten the male chickens of layer strains or to use dual purpose strains. However, there is only limited knowledge on the behavior of these chickens until now. The aim of this study was to test the use of elevated grids and their effect on animal-based indicators (e.g., physical condition). In two successive trials, we kept a total of 1,217 male chickens from three strains (Lohmann Dual, Lohmann Brown Plus, Ross 308) that show differences in growth performance in 24 pens (two trials × three strains × eight pens). In half of the pens, grids were offered at three different heights (enriched groups); in the other half of the pens, no elevated structures were installed (control groups). We recorded the number of birds using the grids at the different heights as well as locomotor activity, walking ability, plumage cleanliness, and the footpad health of chickens. Chickens with low and medium growth performance preferred the highest grids during both the light and dark periods. In contrast, fast-growing chickens used the lowest grid more frequently. Fast-growing chickens kept in the enriched pens tended to have a higher level of locomotor activity and reduced chest cleanliness. Chickens from the medium growth performance strain showed better walking ability when kept in the enriched pens. Enrichment did not affect any of the welfare measures in the slow-growing chickens. These findings suggest that elevated structures may improve chicken welfare, particularly for medium growing chickens. For fast-growing chickens we found evidence for an improvement of animal-based indicators although they used the elevated structures less. However, regardless of growth performance, elevated grids offer the birds an opportunity to rest in a species-specific manner.

Suggestions to Derive Maximum Stocking Densities for Layer Pullets.

Stocking densities for domestic chickens (Gallus gallus domesticus) are regulated by the Council Directives of the European Union for both laying hens and broiler chickens. For layer pullets no regulation of stocking density has been established yet. Based on the existing Council Directives for laying hens (1999/74/EC), broiler chickens (2007/43/EC) and calculations of the floor space that is required for the respective chicken's body, we exemplarily calculated maximum stocking densities for layer pullets. Based on the calculations we obtained absolute additional spaces for birds of different live body mass classes, i.e., useable floor space that the birds have additionally available to the space covered by their body. This allowed us to calculate the relative additional space per individual. We suggest the relative additional space to be a key parameter to derive requirements for a maximum stocking density in layer pullets. We analysed several scenarios for pullets under consideration of the Council Directives for laying hens and for broiler chickens, coming to the conclusion that layer pullets at the end of their rearing period should be provided ideally with a relative additional space of about 40-60%.

Fear but not social behaviour is affected by a polymorphism in the 5'-flanking region of the serotonin transporter (5-HTT) gene in adult hens.

The serotonin transporter gene (5-HTT) is involved in the regulation of the neural serotonin. Polymorphisms in the 5-HTT gene have been described in many species to be involved in physiological processes and emotions. A functional polymorphism in the 5´-flanking region of the 5-HTT gene is known from chickens, with a deletion-allele (D), which is associated with an increased 5-HTT expression, in comparison to the wild-type-allele (W). In domestic populations, the majority of hens carry the W-allele. The regulatory changes of the 5-HTT are accompanied in chickens, as in humans, by modulations of fear. Beside these effects on fear, the understanding of potential functional consequences on the social behaviour in the gregarious chicken is lacking. Thus, we here investigated whether the 5-HTT polymorphism with three genotypes (WW, WD, DD), is not only linked to fear-related behaviour, but affects also socio-positive and -negative behaviours of adult hens. Our data confirmed the effects on fear-related behaviour. WW hens showed highest levels of fear. Interestingly, no differences in the social behaviours were present between the hens of the different 5-HTT genotypes. We further discuss implications for potential evolutionary pathways via natural selection and / or artificial selection through domestication of the 5-HTT polymorphism, which might have enabled a stable social lifestyle in the wild ancestors of modern chickens.

More Than Eggs - Relationship Between Productivity and Learning in Laying Hens.

The intense selection of chickens for production traits, such as egg laying, is thought to cause undesirable side effects and changes in behavior. Trade-offs resulting from energy expenditure in productivity may influence other traits: in order to sustain energetic costs for high egg production, energy expenditure may be redirected away from specific behavioral traits. For example, such energetic trade-offs may change the hens' cognitive abilities. Therefore, we hypothesized highly productive laying hens to show reduced learning performance in comparison to moderate productive lines. We examined the learning ability of four chicken lines that differed in laying performance (200 versus 300 eggs/year) and phylogenetic origin (brown/white layer; respectively, within performance). In total 61 hens were tested in semi-automated Skinner boxes in a three-phase learning paradigm (initial learning, reversal learning, extinction). To measure the hens' learning performance within each phase, we compared the number of active decisions needed to fulfill a learning criteria (80% correct choices for learning, 70% no responses at extinction) using linear models. Differences between the proportions of hens per line that reached criterion on each phase of the learning tasks were analyzed by using a Kaplan-Meier (KM) survival analysis. A greater proportion of high productive hens achieved the learning criteria on each phase compared to less productive hens (Chi2 3 = 8.25, p = 0.041). Furthermore, high productive hens accomplished the learning criteria after fewer active decisions in the initial phase (p = 0.012) and in extinction (p = 0.004) compared to the less selected lines. Phylogenetic origin was associated with differences in learning in extinction. Our results contradict our hypothesis and indicate that the selection for productivity traits has led to changes in learning behavior and the high productive laying hens possessed a better learning strategy compared to moderate productive hens in a feeding-rewarding context. This better performance may be a response to constraints resulting from high selection as it may enable these hens to efficiently acquire additional energy resources. Underlying mechanisms for this may be directly related to differences in neuronal structure or indirectly to foraging strategies and changes in personality traits such as fearfulness and sociality.

Radiographic examination of keel bone damage in living laying hens of different strains kept in two housing systems.

A high prevalence of deviations and fractures of the keel bone is a widespread welfare problem in laying hens. The aim of this study was to experimentally investigate this multifactorial problem throughout the laying period and to compare the prevalence and severity in different layer lines and different housing systems. High performing white (WLA) and brown (BLA) pure bred layer lines and low performing white (R11, G11) and brown layer lines (L68) were kept in both single cages and a floor housing system. A total of 97 hens (19 or 20 from each line, respectively) were repeatedly radiographed in the 35th, 51st and 72nd week of age. Fracture prevalence increased with age (p<0.001). The proportion of deviated keel bone area increased only for caged BLA, WLA and R11 hens (p<0.05) and was significantly higher for caged WLA and R11 hens compared to floor-housed WLA and R11 hens in the 72nd week of age (p<0.05). In the 72nd week of age hens in the floor housing system showed significantly more fractures than hens kept in cages (p<0.05). Prevalence of keel bone deviations was significantly higher in the white layer line R11 but significantly lower in the white layer line G11 compared to both brown layer lines and WLA (p<0.05). Brown layers showed significantly more fractures than white layers (p<0.05) in the 51st and 72nd week of age. Within the brown layers there was a significantly lower prevalence of deviations (p<0.05) and fractures (p<0.05) in the low performing (L68) compared to the high performing line (BLA). Our results show a different development of keel bone damage in caged compared to floor-housed hens under experimental conditions. Additionally, they indicate genetic effects on keel bone damage.

Influence of a Sustained Release Deslorelin Acetate Implant on Reproductive Physiology and Associated Traits in Laying Hens.

The aim of this study was to develop an animal model with non-laying hens which would allow for investigation of the relationship between egg production and common diseases in hens. A total of 40 Lohmann Selected Leghorn hens were kept for 20 weeks in a floor housing system in two groups: group "Adult" (21 weeks of age) and group "Juvenile" (14 weeks of age). In each group, 10 hens were administered a 4.7 mg sustained release deslorelin acetate implant subcutaneously; in group "Adult" after, in group "Juvenile" before the onset of lay. In both groups, the remaining hens served as control hens. An examination of each hen was performed weekly, including ultrasonography to check for ovarian follicles, analysis of estradiol-17ß plasma concentration, and assessment of comb size. Digital radiographs of the keel bone were taken in experimental weeks 7 and 15. No follicles were detected on the ovary of treated hens for a certain time period which varied between individuals (between 8 weeks and until the end of the experiment). Estradiol-17ß concentrations were significantly higher in control hens. The comb was significantly smaller in treated hens. A lower prevalence of keel bone damage (group "Adult") and foot pad dermatitis (FPD) (both groups) was found in treated compared to control hens. These results show that a model with laying and non-laying hens can be achieved by administering a deslorelin acetate implant. Furthermore, they indicate a relationship between egg production and keel bone damage as well as FPD.

Non-specific activation of CD8α-characterised γδ T cells in PBL cultures of different chicken lines.

Avian γδ T lymphocytes still represent an enigmatic cell population not only regarding their functions but also their activation requirements and subset differentiation. To find a suitable method for non-specific stimulation and multiplication of CD8α-characterised γδ T-cell subsets in peripheral blood lymphocyte (PBL) cultures, PBLs of four different chicken lines (WLA, BLA, R11, L68) were treated with a range of commonly used non-specific reagents (PMA, PHA, ConA) and interleukins (IL-2, IL-12, IL-15), and the CD8α(-), CD8α(hi)ß(+) and CD8αα(hi+) γδ T-cell subsets examined in relation to activation (CD25 expression) and proliferation by flow cytometry. The culture of avian PBLs with PMA led to an increase of CD25-expression intensity, whereas IL-2 induced proliferation of γδ T-cells. The combinational application of IL-2 plus PMA functioned synergistically and resulted in significantly enhanced numbers of CD25(+) cells, with simultaneous significant increase of the CD25-expression intensity and proliferation in all γδ T-cell subsets of all chicken lines. The four chicken lines revealed only sporadically differences with frequently highest proliferation rates in PBLs of WLA. In conclusion, PMA in combination with IL-2 is the best suitable additive for avian PBL cultures in order to multiply, activate and maintain CD8α-characterised γδ T-cell subsets of different chicken lines.

Effects of Long-term Graded L-arginine Supply on Growth Development, Egg Laying and Egg Quality in Four Genetically Diverse Purebred Layer Lines.

The present study was conducted to examine effects of long-term graded L-arginine supply on growth development, egg laying and egg quality in four genetically diverse purebred layer lines. The study comprised a rearing trial from hatch to week 16 and a following laying performance trial from week 17 to 41. After hatch 150 one-day-old female chicks of each genotype were distributed to three diets. The experimental diets were equivalent to 70, 100 and 200% L-arginine of age-specific recommended level (National Research Council, 1994) and were offered ad libitum to chicks (hatch to week 7), pullets (week 8 to 16) and hens (week 17 to 41). However, hens' diets were quite low in crude protein. After a pre-laying period from week 17 to 21 thirty-six pullets of each group were used further in the laying performance trial. Independent of chicken's genetic background, insufficient L-arginine supply caused lower body weight, daily weight gain and daily feed intake during the rearing (p<0.001) and induced lower laying intensity and daily egg mass production in the laying period (p<0.05). Parameters fitted to Gompertz function suggested higher adult body weight in L-arginine supplemented birds compared to insufficient supplied ones (p<0.01). Groups fed with insufficient L-arginine reached age of maximum daily weight gain later and showed lowest maximum daily weight gain (p<0.001). As a consequence of limitations in dietary L-arginine and crude protein, high performing genotypes decreased strongly in body weight, daily feed intake and performance compared to the low performing genotypes. In conclusion, L-arginine modified the amount of weight gain and feed intake, especially in growing chicks and pullets independent of genetic background. The high performing hens were more nutritionally stressed than the low performing ones, because concentrations of dietary crude protein were relatively low.

Effects of Graded Dietary L-arginine Supply on Organ Growth in Four Genetically Diverse Layer Lines during Rearing Period.

Little information has been available about the influence of genetic background and dietary L-arginine (Arg) supply on organ growth of chickens. Therefore, the present study examined the effects of a graded ad libitum Arg supply providing 70, 100 and 200% of recommended Arg concentration on organ growth of female chickens from hatch to 18 weeks of age. The chickens derived from four layer lines of different phylogeny (white vs. brown) and laying performance (high vs. low). Based on residual feed and absolute body and organ weights recorded in six-week-intervals, feed consumption, changes of relative organ weights and allometric organ growth were compared between experimental groups. Surplus Arg caused higher feed intake than insufficient Arg (p<0.01) that induced growth depression in turn (p <0.05). During the entire trial chicken's heart, gizzard and liver decreased relatively to their body growth (p<0.001) and showed strong positive correlations among each other. On the contrary, proportions of pancreas and lymphoid organs increased until week 12 (p<0.001) and correlated positively among each other. Due to their opposite growth behaviour (p<0.001), internal organs were assigned to two separate groups. Furthermore, insufficient Arg induced larger proportions of bursa, gizzard and liver compared with a higher Arg supply (p<0.05). In contrast to less Arg containing diets, surplus Arg decreased relative spleen weights (p<0.01). The overall allometric evaluation of data indicated a precocious development of heart, liver, gizzard, pancreas and bursa independent of chicken's genetic and nutritional background. However, insufficient Arg retarded the maturation of spleen and thymus compared with an adequate Arg supply. In conclusion, the present results emphasised the essential function of Arg in layer performance, and indicated different sensitivities of internal organs rather to chicken's dietary Arg supply than to their genetic background.