Dataset Information

An attention bias test to assess anxiety states in laying hens.

ABSTRACT: Fear is a response to a known threat, anxiety is a response to a perceived threat. Both of these affective states can be detrimental to animal welfare in modern housing environments. In comparison to the well-validated tests for assessing fear in laying hens, tests for measuring anxiety are less developed. Perception of a threat can result in an attention bias that may indicate anxious affective states in individual hens following playback of an alarm call. In Experiment 1, an attention bias test was applied to hens that differed in their range access to show that hens that never ranged were more vigilant (stretching of the neck and looking around: P < 0.001) and slower to feed following the second alarm call playback (P = 0.01) compared with hens that ranged daily. All hens showed a reduction in comb temperature following the first alarm call (P < 0.001). In Experiment 2, an open field test was used to determine an effective dose of 2 mg/kg for the anxiogenic drug meta-Chlorophenylpiperazine (m-CPP) in adult laying hens. Hens dosed with 2 mg/kg showed reduced locomotion compared with a saline solution (P < 0.05). In Experiment 3, 2 mg/kg m-CPP or saline was administered to adult hens previously habituated to the open field arena to pharmacologically validate an attention bias test as a measure of anxiety. Hens dosed with m-CPP were slower to feed (P = 0.02) and faster to vocalize following a second alarm call playback (P = 0.03) but these hens did not exhibit the same vigilance behavior as documented in Experiment 1. The m-CPP hens also spent more time stepping and vocalizing (both P < 0.001) than the saline hens. An attention bias test could be used to assess anxiety. However, behavioral responses of hens may vary depending on their age or test environment familiarity, thus further refinement of the test is required. In these tests, 2 mg/kg of m-CPP resulted in motionless behavior when the environment was novel, but more movement and vocalizing when the environment was familiar. The extreme behavioral phenotypes exhibited by individually-tested birds may both be indicators of negative states.

SUBMITTER: Campbell DLM

PROVIDER: S-EPMC6626526 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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An attention bias test to assess anxiety states in laying hens.

Campbell Dana L M DLM Taylor Peta S PS Hernandez Carlos E CE Stewart Mairi M Belson Sue S Lee Caroline C

PeerJ 20190710

Fear is a response to a known threat, anxiety is a response to a perceived threat. Both of these affective states can be detrimental to animal welfare in modern housing environments. In comparison to the well-validated tests for assessing fear in laying hens, tests for measuring anxiety are less developed. Perception of a threat can result in an attention bias that may indicate anxious affective states in individual hens following playback of an alarm call. In Experiment 1, an attention bias tes ...[more]

PMID: 31333910

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Project description:Negative affective states such as anxiety and depression pose a risk to animal welfare, however, practical tests for assessing these states in animals are limited. In humans, anxious individuals are shown to pay more attention toward threatening information than non-anxious individuals, known as an attention bias. Previously, an attention bias test was developed and validated as a measure of anxious states in sheep, where more anxious sheep showed increased attention toward a threat (dog) and were more vigilant than Control animals. Studies in humans suggest that attention biases also occur in depressed individuals, with observations of attention biases toward threats, as well as biases away from positive stimuli. Given these findings, we hypothesized that an attention bias test for sheep could also be used to assess states of depression. We predicted that Merino ewes in pharmacologically induced Depressed (para-chlorophenylalanine) and Anxious (m-chlorophenylpiperazine) states would show greater attention toward a threat than Control animals (saline), but that the Depressed sheep would show relatively less interest in a positive stimulus (photograph of a conspecific). During testing, Depressed sheep paid more attention toward the threat and less toward the photograph than Control animals as predicted (Analyses of Variance, P < 0.05, n = 16 per treatment). Interestingly, Anxious sheep showed an attention bias in the opposite direction, paying more attention toward the photograph and less toward the threat than Control animals (P < 0.05). Both Anxious and Depressed sheep were more vigilant than Control animals (P = 0.002). These results suggest the attention bias test can be used to measure and differentiate states of depression and anxiety in livestock. The bidirectional nature of the attention bias identified between treatments highlights the importance of measuring multiple behaviors in the test and considering the context in which the test is applied. This will enable a clearer characterization of the affective state of an animal, as an aspect of its welfare.

Project description:Maternal stress can affect the offspring of birds, possibly due to hormone deposition in the egg. Additionally, phenotypic diversity resulting from domestication and selection for productivity has created a variety of poultry lines that may cope with stress differently. In this study, we investigated the effects of maternal stress on the behavior of different strains of laying hens and the role of corticosterone as its mediator. For this, fertilized eggs of five genetic lines-two brown (Brown 1 and 2), two white (White 1 and 2), and one pure line White Leghorn-were reared identically as four flocks of 27 birds (24F: 3M) per strain. Each strain was equally separated into two groups: Maternal Stress ("MS"), where hens were subjected to a series of daily acute psychological stressors for 8 days before egg collection, and "Control," which received routine husbandry. Fertile eggs from both treatments were collected at three different ages forming different offspring groups that were treated as replicates; additional eggs from Control were injected either with corticosterone diluted in a vehicle solution ("CORT") or just "Vehicle." Eggs from each replicate were incubated and hatched, and offspring (N = 1,919) were brooded under identical conditions. To measure the effects of maternal stress on anxiety and fear-like behavior, offspring were subjected to a social isolation test (SI) between 5 and 10 days of age and a tonic immobility test (TI) at 9 weeks of age. Compared to Control, MS decreased the number of distress vocalizations emitted by White 2 in SI. No effects of MS were observed in TI, and no effects of CORT were observed in any tests. Overall, brown lines vocalized more in SI and remained in TI for a longer duration than white strains, suggesting genetic differences in fear behavior. Females vocalized more than males in TI and showed a trend toward significance for the same trait in SI. Overall, results suggest that the effects of maternal stress on fearfulness are not directly mediated by corticosterone. Moreover, it highlights behavioral differences across various strains of laying hens, suggesting that fear responses are highly dependent on genotype.

Project description:BackgroundSoya saponin (SS), an active compound in soybean meals, has been widely studied in the medical field. However, it was considered as an anti-nutritional factor in poultry diets. The objective of this experiment was to measure the effects of dietary SS using three dietary treatments on egg-laying performance and immune function of laying hens. Birds were fed a low soybean meal basal diet (CON), a low-SS diet (50 SS) containing 50 mg/kg SS, or a high-SS diet (500 SS) containing 500 mg/kg SS for 10 weeks. At the end of the 5th and 10th week of the trial, samples were collected for analysis.ResultsResults showed that with 50 mg/kg SS supplementation, the egg production rate, feed conversion ratio (FCR), and eggshell quality tended to be improved. Serum follicle stimulating hormone (FSH) and Interleukin-4 (IL-4) levels were also elevated as well as the peripheral blood LPS stimulation index, the proportion of B lymphocytes, and antibody titer of bovine serum albumin (BSA). We also found that mRNA levels of follicle stimulating hormone receptor (FSHR) in ovarian, nuclear transcription factor kappa B (NF-κB), Transforming growth factor (TGF-β) and interferon γ (IFN-γ) in spleen were up-regulated at the end of the trial. Additionally, dietary 50 mg/kg SS improved the ileal flora via up-regulating the relative abundance of Lactobacillus, Romboutsia and Lactobacillus delbrueckii. Although the immune related indicators were improved with 500 mg/kg SS supplemented, it seemed to have a negative influence on the laying-performance. Specifically, serum alanine aminotransferase (ALT), alkaline phosphatase (ALP), and the ratio of IFN-γ to IL-4 were increased in the 500 SS group at the end of the trial. The mRNA levels of gonadotropin releasing hormone 1 (GnRH1) in Hypothalamus, the estrogen related receptor (ERR) in ovaries were downregulated as well as the egg production rate during the trial with 500 mg/kg SS supplemented.ConclusionsThe egg production performance was improved by dietary supplemented with 50 mg/kg SS via increasing ovarian FSHR transcription level and serum estrogen level. A beneficial shift in intestinal microflora was recorded, and the immune function of laying hens was also improved with 50 mg/kg SS supplementation. Surprisingly, the long-term supplementation of 500 mg/kg SS exerted a negative impact on the laying performance and physiological functions of the liver of laying hens.

Project description:Purpose: With the advent of Next-generation sequencing (NGS), several novel genes/proteins and cellular pathways in wide variety of tissues has been discovered. The aim of this study are to perform uterine transcriptome profiling (RNA-seq) to determine differently expressed genes in laying and non-laying hens and to further validate the expression of candidate genes using real-time quantitative reverse transcription polymerase chain reaction (qRT–PCR) in laying, non-laying and molting hens. Methods: Uterine mRNA profiles of 35-60 weeks-old laying and non-laying hens, three each, were generated with NextSeq 500 sequencer in single-end mode with a read length of 1x76 bp. Raw sequencing reads were cleaned and trimmmed with Prinseq tool and good reads were aligned against the chicken reference gemone (Galgal 5.0) in Array Studio. Differential gene expression analysis was performed by the DESeq2 algorithm as implemented in Array Studio. The genes with at least two-fold change (FC) and Benjamini and Hochberg q-value < 0.05 were called differentially expressed. Results: Using an optimized data analysis workflow, we mapped about 32 million reads from layers and 28 million reads from non-layers to the chicken genome. A total of 19,152 gene transcripts were annotated from Ensembl alignment which represents 50.24% of the chicken genome assembly. Differential gene expression analysis showed 616 were differentially expressed between layer and non-layer hens. 229 DEGs were significantly up-regulated and 286 were significantly down-regulated in the laying hens when compared to the non-laying hens. Twelve candiate genes, linked to calcium remodeling, were identified by gene function analysis and validated using qPCR. MEPE, CALCB, OTOP2, STC2 and ATP2C2 were confirmed to be highly expressed in laying hens as compared to molting and non-laying hens. RNA-seq and qPCR data for relative gene expression were highly correlated (R2 =0.99). Conclusions: Our study reports the expression of four novel genes that are speculated to transport calcium ions across the uterine epithellium for eggshell mineralization. These genes can be used as quantitative basis of selecting hens with an improved eggshell quality.

Dataset Information

An attention bias test to assess anxiety states in laying hens.

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An attention bias test to assess anxiety states in laying hens.

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