Effects of simple cage enrichment and its removal on the behavior and welfare of American mink (Neogale vison).

Environmental enrichment may reduce stereotypies in fur-farmed mink. North American mink standards require manipulable enrichment objects within cages. However, mink can rapidly destroy objects inhibiting continuous enrichment presence, which may have negative welfare impacts. This experimental study determined the effects of removing simple cage manipulable enrichments (plastic chains and dumbbells), either short-term or longer-term, on the behavioral expression of welfare in fur-farmed mink. Locomotor stereotypies, normal activity, sub-types of inactivity related to boredom, and tail fur-chewing were recorded across four treatment groups with either (1) no enrichment, (2) continuous enrichment, (3) short (temporary), or (4) long-term enrichment removal. Contrary to predictions, locomotor stereotypies, and scrabbling were not reduced by the enrichments, nor affected by the enrichment removal. Observations at the beginning of the trial showed the non-enriched mink spent the least amount of time lying with their eyes open (i.e., the least bored). The lack of enrichment may have increased fur-chewing on the tail, but larger sample sizes would be needed for statistical confirmation. This research contributes to the literature on evaluating simple, practical enrichments for improving fur-farmed mink welfare.

Loss of a grooming enrichment impacts physical, behavioural, and physiological measures of welfare in grazing beef cattle.

Pasture-based beef cattle are raised in a range of production environments. Some paddocks may contain trees and other objects that allow for grooming, hence being naturally enriching, whilst others may be barren without these opportunities. Additionally, it is not uncommon for cattle to move between these enriched and barren environments as part of routine management. While the benefits of enrichment are well studied, how this 'enrichment loss' impacts cattle welfare as access to stimuli is removed is unknown. This trial assessed the impacts of the loss of an enriching object (grooming brush) on grazing beef cattle welfare and production characteristics. When grooming brush access was blocked, cattle became dirtier, showed reduced average daily gain, and had elevated faecal cortisol metabolites, although this varied according to the degree of initial individual brush use. Additionally, allogrooming and grooming on other objects were reduced when access to the brush was returned, potentially indicating a rebound effect. These results demonstrate that the loss of adequate grooming objects can impair the overall welfare of grazing cattle; however, further work is needed to determine exactly which natural or artificial objects provide adequate grooming opportunities.

Sex impacts pain behaviour but not emotional reactivity of lambs following ring tail docking.

Studies in humans have shown sex differences in response to painful events, however, little is known in relation to sex differences in sheep. Understanding sex differences would enable improved experimental design and interpretation of studies of painful procedures in sheep. To examine sex differences in response to pain, 80 lambs were tested across five cohorts of 16. The lambs were penned in groups containing two male and two female lambs with their respective mothers. Lambs were randomly allocated from within each block to one of four treatment groups; FRing-Female lamb, ring tail docked without analgesia, MRing-Male lamb, ring tail docked without analgesia, FSham-Female lamb, tail manipulated and MSham-Male lamb, tail manipulated. Following treatment, lambs were returned to their pen and were video recorded for 45 mins for behavioural observations of acute pain and posture. An hour after treatment, lambs then underwent an emotional reactivity test that consisted of three phases: Isolation, Novelty and Startle. Following treatment, Ring lambs displayed more abnormal postures (mean = 2.5 ± 0.5) compared to Sham lambs (mean = 0.05 ± 0.4, P = 0.0001). There was an effect of sex on the display of acute pain-related behaviours in lambs that were tail docked (P < 0.001), with female lambs displaying more acute behaviours (mean count = +2.2). This difference in behaviour between sexes was not observed in Sham lambs. There was no effect of sex on display of postures related to pain (P = 0.99). During the Novelty and Startle phase of the emotional reactivity test, Ring lambs tended to (P = 0.084) or did (P = 0.018) show more fear related behaviours, respectively. However, no effect of sex was observed. The results of this study indicate that a pain state may alter the emotional response of lambs to novel objects and potential fearful situations. It was also demonstrated that female lambs display increased sensitivity to the acute pain caused by tail docking compared to males.

Minimal effects of ultraviolet light supplementation on egg production, egg and bone quality, and health during early lay of laying hens.

Chicken vision is sensitive to ultraviolet (UV) light containing the UVA spectrum, while UVB plays a key role in the endogenous production of vitamin D3. However, commercially available light sources are typically deficient in the UV spectrum and thus may not adequately fulfill the lighting requirements of indoor-housed laying hens. We hypothesized that supplementary UVB light may improve egg production and egg quality, and bone health during early lay relative to UVA supplementation or standard control lighting. To investigate the effects of UV light supplementation, an experiment was conducted on 252 ISA Brown hens during 16 to 27 weeks of age. Birds were housed in eighteen pens (14 hens/pen) under three different light treatment groups each with six replications: (i) UVO: standard control lighting with LED white light, (ii) UVA: control lighting plus supplemental daylight with an avian bulb, and (iii) UVA/B: control lighting plus a supplemental full spectrum reptile bulb containing both UVA and UVB wavelengths. Hen-day egg production and egg quality, blood parameters including plasma Ca and P, and serum 25(OH)D3, and hen body weight and external health scoring were measured at different age points; while bone quality was assessed at the end of the experiment at 27 weeks. Data were analyzed in JMP® 16.0 using general linear mixed models with α level set at 0.05. Results showed that UVA and UVA/B supplemented birds reached sexual maturity (50% production) 3 and 1 day earlier, respectively, than control birds. There was a trend for UV lights to increase hen-day egg production (P = 0.06). Among egg quality traits, only eggshell reflectivity and yolk index were affected by UV lights (P = 0.02 and 0.01, respectively); however, most of the egg quality traits changed over age (all P < 0.01). Post-hoc tests showed higher serum 25(OH)D3 in the UVA/B group relative to control hens (P < 0.05); but there was no treatment effect on plasma Ca and P or on bone quality parameters (all P > 0.05). A significant interaction was observed between light treatment and age for the number of comb wounds (P = 0.0004), with the UV supplemented hens showing more comb wounds after 24 weeks. These results demonstrated that supplemental UVA/B light had minimal effects on egg production and egg quality, whereas, UVA/B exposure may increase vitamin D3 synthesis during the early laying period. The optimum duration of exposure and level of intensity needs to be determined to ensure these benefits.

Shared stressful experiences affect social proximity in Merino sheep.

While it is well established that humans develop stronger relationship bonds when they share stressful experiences, there is little known on how shared stressful experiences may influence relationship bonding in animals. Here, we present a study looking at social proximity between individuals in small groups of Merino ewes following a shared stressful experience compared with control sheep that were not exposed to stress. Some sheep were familiar to each other. Analyses of social proximity using real-time-kinematic Global Navigation Satellite System (GNSS) on-animal devices showed sheep preferred to be closest to familiar individuals, but across the study duration they also developed a preference for the individuals they shared the stressful experience with, relative to their proximity to control individuals. These results contribute to limited research on what factors may instigate the development of bonds between unfamiliar sheep. Between-individual bonds may develop as a means of socially mediated stress buffering. Social bonding following a shared stressful experience aligns with human social relationships and increases our understanding of how animals perceive their conspecifics in relation to stressful environmental change.

Automated Virtual Fencing Can Effectively Contain Sheep: Field Trials and Prospects.

Virtual fencing technology uses on-animal devices to communicate boundaries via a warning audio tone and electrical pulse signals. There is currently limited validation work on sheep. This study used modified cattle eShepherd® virtual fencing neckbands on reduced-wool sheep with clipped necks to enable automated trials with small groups across both day and night. The first 5-day trial with six Dorper crossbred sheep was conducted in an experimental paddock setting, with a second 5-day trial conducted with 10 Ultra White sheep on a commercial farm. The animals across both trials were contained in the inclusion zone for 99.8% and 92.2% of the trial period, with a mean percentage (±SD) of total audio cues as audio only (i.e., not followed by an electrical pulse) being 74.9% ± 4.6 in the first trial, and 83.3% ± 20.6 for the second trial. In the second trial, sheep crossed over into the exclusion zone on the third night and remained there until they were walked out for their daily yard check in the morning. These preliminary trial results are promising for the use of automated technology on sheep, but suitable devices and algorithms still need to be designed specifically for sheep in the long term.

Beef Cattle Preference and Usage of Environmental Enrichments Provided Simultaneously in a Pasture-Based Environment.

Environmental enrichment can improve livestock welfare through increasing environmental complexity to promote a greater range of natural behaviours. However, there is limited understanding of the need for and impacts of enrichments for extensively managed beef cattle that can sometimes be kept in grassed paddocks devoid of additional natural and artificial features, i.e., 'barren pastures'. This trial assessed which enrichments beef cattle preferred and utilised in a barren paddock environment. Eight groups of seven Angus steers housed on pastured paddocks devoid of natural or artificial features were observed during daylight hours for two days a week over a period of three weeks, after being presented with four enrichments simultaneously: a cattle brush, a piece of hanging rope, a tree stump, and a woodchip pile. Although enrichment use generally decreased over time, the brush, stump, and woodchip maintained a higher level of use than the rope, based on the frequency of interactions and number of displacements around the enrichments (both p < 0.001). This suggests that the brush, stump, and woodchip pile were more valuable resources to the cattle, allowing for grooming and lying behaviours, although oral manipulations also occurred on the stump, woodchip, and rope. The inclusion of these enrichments can increase the complexity of barren pasture environments and allow for the increased expression of natural behaviours, potentially contributing to improved welfare.

Behavior and welfare impacts of water provision via misting in commercial Pekin ducks.

Ducks will access water to maintain feather condition and exhibit natural water-related behaviors such as wet preening. Providing water to ducks commercially is challenging as it may reduce litter and air quality leading to higher duck mortality or illness. This research aimed to measure the behavioral and welfare impacts of water provision via a misting system for commercial Pekin grower ducks in Victoria, Australia. Seven grower flocks were observed (four misted and three nonmisted in open-sided sheds) during May and November 2021. From 26 until 33 d of age, treatment ducks were provided 1 h of misting with shed curtains closed in both treatment and control sheds. At the start and end of the misting application period, external health and welfare measures were taken directly on the ducks via transect walks throughout each shed and catch-and-inspect observations on a sample of 150 ducks from each shed. Video recordings were also made of the misted and nonmisted ducks for 3 h representing time periods prior to, during, and after the 1-h misting across all sheds for all 8 d of the treatment period. Observations were made of all behavior that ducks exhibited at 10-min scan sample intervals across four cameras per shed, totaling 4,198 scans across the seven sheds. General linear mixed models showed the misting application predominantly had impacts on the patterns of behavioral change across the treatment time periods between the misted and nonmisted ducks rather than increasing or decreasing the overall expression of specific behaviors (interaction terms all P ≤ 0.003). The misted ducks increased drinking, tail wagging, and walking, and reduced preening, rooting litter, sitting, and stretching during misting relative to what they showed prior. The nonmisted ducks showed less sitting and more panting during misting relative to prior. Pearson's Chi-square tests showed some differences between the treatment groups in feather cleanliness on the back and wings (both P < 0.0001), likely resulting from pre-existing differences between sheds in blood from pin feathers. Most welfare indicators showed no positive or negative effect of the misting treatment. These results indicate overhead misting does affect duck behavior to some degree without compromising their welfare, but further research with larger water droplet sizes resulting in greater accumulation of surface water or extended durations of misting may lead to greater effects.

Ducks use bathing water for wet preening and feather maintenance. Commercially, it is challenging to provide clean bathing water without compromising litter quality and duck health. Overhead misting may be a mode of water delivery that will wet the ducks' bodies with continuously clean water. This study compared seven grower flocks of Pekin ducks (four misted and three nonmisted treatment flocks) in open-sided sheds during May and November 2021 in Australia. From 26 until 33 d of age, treatment ducks were provided 1 h of misting with shed curtains closed in both treatment and control sheds. External welfare measures were taken directly on the ducks at 26 and 33 d of age. Daily video recordings were made to observe if behaviors differed before, during, or after the 1 h of misting in both treatment and control sheds. Results showed the misting application predominantly affected the way behaviors changed across time between the misted and nonmisted ducks rather than increasing or decreasing the overall expression of specific behaviors. The differences may have in part been related to the curtain closure. Most welfare indicators showed no positive or negative effect of the misting treatment. Larger water droplet sizes may have greater effects on duck behavior.

Relationship between sunlight and range use of commercial free-range hens in Australia.

In Australia, summer brings intense, bright sunlight with high ultraviolet (UV) radiation and hot temperatures, which might impact free-range hens' ranging outside. To determine how range use was correlated with different sunlight variables and weather factors, a study was carried out on three commercial free-range layer farms during the summer/autumn period (December-May) across diverse regions of Australia in Tasmania (TAS), Queensland (QLD), and Western Australia (WA). Hens' range distribution was determined by counting the number of hens in the direct sunlight ('sun') or 'cloud' and shaded areas ('sun-shade' or 'cloud-shade', in sunny or cloudy conditions, respectively) using image snapshots taken at 30 min intervals from video recordings of a portion of one shed comprising 20,000-30,000 hens on each farm during the production phase of the laying cycle. The solar radiation spectrum [UV radiation (UVAB) (288-432 nm), photosynthetically active radiation (PAR; visible light) (400-700 nm), and total solar radiation (TSR) (285 nm-3000 nm)] and weather data (ambient temperature and relative humidity) were recorded through an on-site weather station. Data were analysed separately due to discrepancies between the farms' layouts. The effects of time of day and months on range use were analysed using General Linear Models in JMP® 16.0 and the relationship of sunlight and weather variables with hens' distribution in 'sun'/'sun-shade' and 'cloud'/'cloud-shade' in sunny and cloudy conditions respectively was determined by fitting linear ridge regression models using the 'lmridge' package in R. Overall, the time of day and month had significant effects on hens' distribution on the range (all p < 0.0001). Hens' range occupancy in the 'sun' decreased during the midday period with gradual increases in the late afternoon to evening, and the opposite pattern in the 'sun-shade'. A linear increase in the number of hens on the range over the months indicated the seasonal effects on hen ranging patterns. Temperature, UVAB and PAR were the most important factors for discouraging hens' range use in the 'sun' suggesting free-range systems in Australia should be designed to account for the extreme sunlight using adequate shade for optimum ranging across summer.

Impacts of Rearing Enrichments on Pullets' and Free-Range Hens' Positive Behaviors across the Flock Cycle.

Enrichment during the indoor rearing of pullets destined for free-range systems may optimize pullet development including increasing motivated natural behaviors (termed 'positive behaviors') including foraging, dust bathing and chick play. Hy-Line Brown® chicks (n = 1700) were floor-reared indoors across 16 weeks with three enrichment treatments (n = 3 pens/treatment): (1) standard control, (2) weekly novel objects-'novelty', (3) perching/navigation structures-'structural'. At 16 weeks, pullets (n = 1386) were transferred to nine identical pens within rearing treatments with outdoor range access from 25 to 65 weeks. Video cameras recorded the pullet pens, adult indoor pens, and outside range. During rearing, observations of play behavior (running, frolicking, wing-flapping, sparring) in chicks at 2, 4 and 6 weeks (total of 432 thirty-second scans: 16 observations × 3 days × 9 pens) showed no overall effect of rearing treatment (p = 0.16). At 11 and 14 weeks only the 'novelty' hens were observed to increase their foraging across age (p = 0.009; dust bathing: p = 0.40) (total of 612 thirty-second scans per behavior: 17 observations × 2 days × 2 age points × 9 pens). Observations of adult hens at 26, 31, 41, 50, 60 and 64 weeks showed that the structural hens exhibited overall more dust bathing and foraging than the control hens (both p < 0.04) but both novelty and/or structural hens showed small increases depending on the behavior and location (total of 4104 scans per behavior: 17 observations × 2 days × 6 age points × 9 pens × 2 locations = 3672 + an additional 432 observations following daylight saving). Across age, adult hens differed in the degree of dust bathing performed inside or outside (both p ≤ 0.001) and foraging outside (p < 0.001) but not inside (p = 0.15). For litter-reared pullets, additional enrichments may result in some long-term increases in positive behaviors.

Commercial Free-Range Laying Hens' Preferences for Shelters with Different Sunlight Filtering Percentages.

Extreme sunlight might be aversive to free-range laying hens, discouraging them from going outside. Range enrichment with artificial shelters may protect hens from sunlight and increase range use. The preferences of 34-40-week-old Hy-Line Brown laying hens for artificial shelters were assessed by counting the number of hens under three densities of individual shelters (three replicates/density) from video recordings for 14 to 17 days for two flocks. The artificial shelters used shade cloth marketed as blocking 50%, 70%, and 90% of ultraviolet light, although other sunlight wavelengths were also reduced. Different sunlight spectral irradiances (ultraviolet radiation (UVAB) (288-432 nm), photosynthetically active radiation (PAR) (400-700 nm), and total solar radiation (TSR) (285 nm-3000 nm), ambient temperature, and relative humidity were recorded with an on-site weather station. There was a significant interaction between sunlight-filtering shelter and time of day (both Flocks, p < 0.0001), i.e., hens preferred shelters with the highest amount of sunlight-filtering at most time points. Regression models showed that the most variance in shelter use throughout the day resulted from the ambient temperature in both flocks, while sunlight parameters had different degrees of effect depending on the shelter type and flock. However, fewer hens under the shelters during the midday period suggest that during periods of intense sunlight, hens prefer to remain indoors, and artificial structures might not be sufficient to attract more hens outside.

A Perspective on Strategic Enrichment for Brain Development: Is This the Key to Animal Happiness?

Livestock animals are sentient beings with cognitive and emotional capacities and their brain development, similar to humans and other animal species, is affected by their surrounding environmental conditions. Current intensive production systems, through the restrictions of safely managing large numbers of animals, may not facilitate optimal neurological development which can contribute to negative affective states, abnormal behaviors, and reduce experiences of positive welfare states. Enrichment provision is likely necessary to enable animals to reach toward their neurological potential, optimizing their cognitive capacity and emotional intelligence, improving their ability to cope with stressors as well as experience positive affect. However, greater understanding of the neurological impacts of specific types of enrichment strategies is needed to ensure enrichment programs are effectively improving the individual's welfare. Enrichment programs during animal development that target key neurological pathways that may be most utilized by the individual within specific types of housing or management situations is proposed to result in the greatest positive impacts on animal welfare. Research within livestock animals is needed in this regard to ensure future deployment of enrichment for livestock animals is widespread and effective in enhancing their neurological capacities.

Preference testing for UV light spectrum and intensity in laying hens.

Sunlight intensity and UV radiation may affect free-range hens' use of the outside range, particularly when sunlight is intense with a high UV index. However, it is uncertain what aspect of sunlight (brightness or UV) may be most aversive to hens to discourage them from leaving standard indoor lighting conditions to venture outdoors. A controlled indoor-based choice study was conducted to determine whether hens showed preferences for different light wavelengths and intensities that may affect outdoor range usage. Cage-reared ISA Brown laying hens (n = 84) at 44 wk of age in 3 groups (28 hens/group) were tested for preferences of indoor standard light emitting diode (LED) white light (control) vs. one of three different treatment lights: 1) visible spectrum plus infrared wavelengths (VIS); 2) visible spectrum plus UVA wavelengths (UVA); and (iii) visible spectrum plus UVA and UVB wavelengths (UVA/B) presented successively at low, medium, or high levels of intensity. Hens within each group were individually tested for 2 h in an apparatus with 2 compartments (control vs. treatment) connected by a tunnel on both sides. Videos of hens' time spent in each compartment and behaviors were decoded and analyzed using GLMM. Hens spent more time under the low intensity of the UVA/B light treatment (62%), the low intensity of VIS light (61%), medium intensities of both UVA/B light (60%), and UVA light (59%), and the high intensity of the VIS light (58%) when compared with control light (all P ≤ 0.05). Hens spent less time feeding under all intensities of UVA light (all P ≤ 0.03) and showed more foraging, ground pecking, and preening at lower levels of UVA/B light (P < 0.05). The study suggests that UVA/B light (sunlight) may have positive effects for hen range use, but during peak sun intensities, hens may need additional measures (e.g., shelter) to protect themselves. Confirmation of these findings in a free-range setting is needed.

A Multi-Disciplinary Approach to Assess the Welfare Impacts of a New Virtual Fencing Technology.

Virtual fencing involving the application of audio cues and electrical stimuli is being commercially developed for cattle. Virtual fencing has the potential to improve productivity through optimized pasture management and utilization by grazing animals. The application of virtual fencing initiates public concern for the potential welfare impacts on animals due the aversive nature of using an electrical stimulus. It is therefore important to provide welfare assurance of the impacts of virtual fencing on livestock. In this paper, we provide an overview of the welfare assessment and validation stages for virtual fencing which could be applied to other new technologies utilizing novel systems. An understanding of stress measures and their suitability for use in specific contexts is discussed, including the use of glucocorticoids to measure both acute and chronic stress, and behavioral responses and patterns to indicate welfare states. The importance of individual differences in relation to learning and cognition are also highlighted. Together, this multi-disciplinary approach to welfare assessment provides a tool kit that may be applied for welfare assurance of some new technologies and systems for farm animals.

Validation of Real-Time Kinematic (RTK) Devices on Sheep to Detect Grazing Movement Leaders and Social Networks in Merino Ewes.

Understanding social behaviour in livestock groups requires accurate geo-spatial localisation data over time which is difficult to obtain in the field. Automated on-animal devices may provide a solution. This study introduced an Real-Time-Kinematic Global Navigation Satellite System (RTK-GNSS) localisation device (RTK rover) based on an RTK module manufactured by the company u-blox (Thalwil, Switzerland) that was assembled in a box and harnessed to sheep backs. Testing with 7 sheep across 4 days confirmed RTK rover tracking of sheep movement continuously with accuracy of approximately 20 cm. Individual sheep geo-spatial data were used to observe the sheep that first moved during a grazing period (movement leaders) in the one-hectare test paddock as well as construct social networks. Analysis of the optimum location update rate, with a threshold distance of 20 cm or 30 cm, showed that location sampling at a rate of 1 sample per second for 1 min followed by no samples for 4 min or 9 min, detected social networks as accurately as continuous location measurements at 1 sample every 5 s. The RTK rover acquired precise data on social networks in one sheep flock in an outdoor field environment with sampling strategies identified to extend battery life.

Relationship between Range Use and Fearfulness in Free-Range Hens from Different Rearing Enrichments.

Inconsistency between the environments of indoor pullet rearing and adult outdoor housing may increase the fearfulness in free-range hens. Rearing enrichments and/or range use may reduce adult fearfulness. Hy-Line Brown® chicks (n = 1700) were reared inside across 16 weeks with three enrichment treatments: weekly changing novel objects, custom-designed perching/navigation structures, or no additional enrichments. Pullets were transferred to a free-range system at 16 weeks of age, with range access provided from 25 weeks. At 62 weeks, 135 hens were selected from the three rearing treatments and two ranging groups (indoor: no ranging and outdoor: daily ranging) based on individual radio-frequency identification tracking. Individual behavioural tests of tonic immobility, emergence, open field, and novel object (pen level) were carried out on hens. Spectrograms of vocalisations were analysed for the open field test, as well as computer vision tracking of hen locomotion. The results showed few effects of rearing treatments, with outdoor rangers less fearful than indoor hens. The latency to step in the open field test negatively correlated with hen feather coverage. These results show that individual variation in ranging behaviours is present even following rearing enrichment treatments, and subsequent range use might be an indicator of bird fearfulness.

Cecal Microbiota of Free-Range Hens Varied With Different Rearing Enrichments and Ranging Patterns.

Free-range pullets are reared indoors but the adult hens can go outside which is a mismatch that may reduce adaptation in the laying environment. Rearing enrichments might enhance pullet development and adaptations to subsequent free-range housing with impact on behavior and health measures including gut microbiota. Adult free-range hens vary in range use which may also be associated with microbiota composition. A total of 1,700 Hy-Line Brown® chicks were reared indoors across 16 weeks with three enrichment treatment groups: "control" with standard litter housing, "novelty" with weekly changed novel objects, and "structural" with custom-designed perching structures in the pens. At 15 weeks, 45 pullet cecal contents were sampled before moving 1,386 pullets to the free-range housing system. At 25 weeks, range access commenced, and movements were tracked via radio-frequency identification technology. At 65 weeks, 91 hens were selected based on range use patterns ("indoor": no ranging; "high outdoor": daily ranging) across all rearing enrichment groups and cecal contents were collected for microbiota analysis via 16S rRNA amplicon sequencing at V3-V4 regions. The most common bacteria in pullets were unclassified Barnesiellaceae, Prevotella, Blautia and Clostridium and in hens Unclassified, Ruminococcus, unclassified Lachnospiraceae, unclassified Bacteroidales, unclassified Paraprevotellaceae YRC22, and Blautia. The microbial alpha diversity was not significant within the enrichment/ranging groups (pullets: P ≥ 0.17, hen rearing enrichment groups: P ≥ 0.06, hen ranging groups: P ≥ 0.54), but beta diversity significantly varied between these groups (pullets: P ≤ 0.002, hen rearing enrichment groups: P ≤ 0.001, hen ranging groups: P ≤ 0.008). Among the short-chain fatty acids (SCFAs), the propionic acid content was higher (P = 0.03) in the novelty group of pullets than the control group. There were no other significant differences in the SCFA contents between the rearing enrichment groups (all P ≥ 0.10), and the ranging groups (all P ≥ 0.17). Most of the genera identified were more abundant in the indoor than high outdoor hens. Overall, rearing enrichments affected the cecal microbiota diversity of both pullets and adult hens and was able to distinguish hens that remained inside compared with hens that ranging daily for several hours.