Comparison of physiological markers, behavior monitoring, and clinical illness scoring as indicators of an inflammatory response in beef cattle.

Clinical illness (CI) scoring using visual observation is the most widely applied method of detecting respiratory disease in cattle but has limited effectiveness in practice. In contrast, body-mounted sensor technology effectively facilitates disease detection. To evaluate whether a combination of movement behavior and CI scoring is effective for disease detection, cattle were vaccinated to induce a temporary inflammatory immune response. Cattle were evaluated before and after vaccination to identify the CI variables that are most indicative of sick cattle. Respiratory rate (H2 = 43.08, P < 0.0001), nasal discharge (H2 = 8.35, P = 0.015), and ocular discharge (H2 = 16.38, P = 0.0003) increased after vaccination, and rumen fill decreased (H2 = 20.10, P < 0.0001). Locomotor activity was measured via leg-mounted sensors for the four days preceding and seven days following vaccination. A statistical model that included temperature, steps, lying time, respiratory rate, rumen fill, head position, and excess saliva was developed to distinguish between scores from before and after vaccination with a sensitivity of 0.898 and specificity of 0.915. Several clinical illness signs were difficult to measure in practice. Binoculars were required for scoring respiratory rate and eye-related metrics, and cattle had to be fitted with colored collars for individual identification. Scoring each animal took up to three minutes in a small research pen; therefore, technologies that can automate both behavior monitoring and identification of clinical illness signs are key to improving capacity for BRD detection and treatment.

Dogs' ability to detect an inflammatory immune response in cattle via olfaction.

Introduction: Canine olfaction is a potential means for detection of respiratory disease in beef cattle. In a prior study, two dogs were trained to discriminate between nasal swabs from healthy cattle and cattle that developed Bovine Respiratory Disease. Dogs had some ability to identify samples from BRD-affected cattle, but results were ambiguous. The purpose of this study was to evaluate more dogs using better-controlled training and testing procedures. Methods: Nasal and saliva swabs were collected from 96 cattle before and after administering a vaccine to induce an inflammatory immune response. Samples were stored at -80°C for up to 11 months before use, and samples from animals with an elevated body temperature at baseline were omitted. An automated olfactometer apparatus was constructed to improve blinding procedures and reduce opportunities for odor contamination. Four dogs were trained to distinguish between swabs from healthy and sickness-model cattle, including the two dogs from the previous study ("Runnels" and "Cheaps") and two inexperienced dogs ("Molokai" and "Amy"). During a seven-month training period, dogs were exposed to samples from 28 animals. Dogs were tested on 59 sets of unfamiliar samples. Results: Performance varied among dogs (χ2 = 10.48, p = 0.02). Molokai's performance was above chance (0.73 ± 0.06, p = 0.0006), while Amy (0.44 ± 0.06, p = 0.43), Cheaps (0.53 ± 0.07, p = 0.79), and Runnels (0.56 ± 0.06, p = 0.43) did not respond correctly at a rate different from chance. Accuracy did not differ between nasal swabs (0.63 ± 0.08) and saliva swabs (0.53 ± 0.08, χ2 = 0.81, p = 0.37). Discussion: The results of this study indicate that canine olfaction may be an effective means of detecting illness in beef cattle. However, individual dogs' aptitude for this detection task varies.

Current challenges and future of agricultural genomes to phenomes in the USA.

Dramatic improvements in measuring genetic variation across agriculturally relevant populations (genomics) must be matched by improvements in identifying and measuring relevant trait variation in such populations across many environments (phenomics). Identifying the most critical opportunities and challenges in genome to phenome (G2P) research is the focus of this paper. Previously (Genome Biol, 23(1):1-11, 2022), we laid out how Agricultural Genome to Phenome Initiative (AG2PI) will coordinate activities with USA federal government agencies expand public-private partnerships, and engage with external stakeholders to achieve a shared vision of future the AG2PI. Acting on this latter step, AG2PI organized the "Thinking Big: Visualizing the Future of AG2PI" two-day workshop held September 9-10, 2022, in Ames, Iowa, co-hosted with the United State Department of Agriculture's National Institute of Food and Agriculture (USDA NIFA). During the meeting, attendees were asked to use their experience and curiosity to review the current status of agricultural genome to phenome (AG2P) work and envision the future of the AG2P field. The topic summaries composing this paper are distilled from two 1.5-h small group discussions. Challenges and solutions identified across multiple topics at the workshop were explored. We end our discussion with a vision for the future of agricultural progress, identifying two areas of innovation needed: (1) innovate in genetic improvement methods development and evaluation and (2) innovate in agricultural research processes to solve societal problems. To address these needs, we then provide six specific goals that we recommend be implemented immediately in support of advancing AG2P research.

Consider the Source: The Impact of Social Mixing on Drylot Housed Steer Behavior and Productivity.

Cattle are a social species in which social mixing can induce physical and psychosocial stress; however, the impact of social mixing on cattle welfare is unknown. Two different sources of genetically similar Angus crossbred steers were transported to the same feedlot and assigned to a pen where they were either socially mixed or housed with individuals from their source herds. Social mixing did not impact average daily gains in pens, feed intake, or feed efficiency; pens of socially mixed steers were more active. Sources differed in their responses to social mixing. One source was unaffected, whereas social mixing negatively impacted productivity for the other source. Irrespective of social mixing, the sources differed in the amount of time per day they spent ruminating and drinking. Group analyses indicated that socially mixing two sources of feedlot steers did not negatively impact group productivity, yet the impacts that were observed at the individual level suggest that prior experiences may influence their ability to cope with social stress, emphasizing the importance of early-life experiences to long-term welfare and productivity. Social mixing was not universally detrimental to cattle welfare, and the source of cattle may have the greatest affect on their performance regardless of whether a social mixing event has occurred.

Evaluating accurate and efficient sampling strategies designed to measure social behavior and brush use in drylot housed cattle.

Efficient sampling strategies expedite behavioral data collection. While multiple studies have evaluated sampling strategies for core behaviors in cattle, few have focused on social interactions. To identify sampling strategies that accurately captured cattle social behaviors and brush use feedlot steers (n = 3 pens; 9 steers/pen) were observed from 8:00 to 17:00. Average bout duration (sec), total duration per day (sec), and bout frequency were recorded for allogrooming, bar licking, tongue rolling, and brush utilization. Frequency was recorded for headbutting and mounting. Data was extracted from continuous observation datasets using eight different sampling strategies and the results subsequently compared. Differences among sampling strategies were evaluated using a non-parametric One-Way ANOVA Kruskal-Wallis Test. Pearson correlation evaluated the strength of association between a specific sampling strategy and continuous observations. Bout duration for allogrooming (P > 0.65), bar licking (P > 0.60), tongue rolling (P > 0.99), brush use (P > 0.99), and mounting frequency (P > 0.70) did not differ from continuous observations. Tongue rolling (r2 > 0.95, P <0.0001) and brush use (r2 > 0.70, P < 0.0003) were best captured when cattle were observed from 08:00 to 14:00. When cattle were continuously observed from 08:00 to 14:00 or for 15 minutes every 30 minutes, allogrooming (P > 0.2) (frequency, duration), bar licking (P > 0.95) (frequency, duration), brush use (P > 0.1) (frequency, duration), heat butt (P > 0.30) (frequency), or tongue rolling (P > 0.30) (frequency, duration) did not differ from continuous observations. Observing cattle for 15 minutes every 30 minutes yielded the highest accuracy for all behavioral metrics and was considered the most effective strategy for comprehensively evaluating cattle social behavior (r2 > 75; P < 0.05). These results provide insight into accurate and efficient sampling strategies that expedite social behavior data collection in cattle and will facilitate efficient generation of new knowledge regarding cattle social behaviors.

Effect of Dietary Corn Stalk Inclusion on the Performance of Non-Nutritive Oral Behaviors of Drylot-Housed Beef Steers.

Dietary forage levels contribute to the performance of non-nutritive oral behaviors (NNOB) in cattle, yet the impact of varying forage levels on these behaviors is unknown. To evaluate the impact of dietary corn stalk inclusion (CSI) levels on NNOB, rumination time, and activity, pre-dominantly British-continental crossbred drylot-housed steers (n=27) were blocked by weight and randomly assigned to one of three dietary treatments (5%, 10%, or 15%) of CSI on a DM basis. Animals were fitted with a rumination collar upon arrival that measured rumination time and activity and video recorded. Cattle that spent more time bar licking had greater DMI, tended to have greater ADG and be more active. CSI in this study did influence NNOB performance; however, the impacts observed were not as expected. Cattle fed the 10% CSI performed the most bar licking and tongue rolling. This pilot investigation suggest that these CSI were insufficient to have a meaningful impact on NNOBs. Cattle spending more time bar licking and bar licked more frequently may be more orally motivated as reflected in their increased DMI and activity levels.

Using Canine Olfaction to Detect Bovine Respiratory Disease: A Pilot Study.

Bovine respiratory disease (BRD) is the leading cause of morbidity and mortality in feedlot cattle and is a major welfare and economic concern. Identification of BRD-affected cattle using clinical illness scores is problematic, and speed and cost constraints limit the feasibility of many diagnostic approaches. Dogs can rapidly identify humans and animals affected by a variety of diseases based on scent. Canines' olfactory systems can distinguish between patterns of volatile organic compounds produced by diseased and healthy tissue. In this pilot study, two dogs ("Runnels" and "Cheaps") were trained for 7 months to discriminate between nasal swabs from cattle that developed signs of BRD within 20 days of feedlot arrival and swabs from cattle that did not develop BRD signs within 3 months at the feedlot. Nasal swabs were collected during cattle processing upon arrival to the feedlot and were stored at -80°C. Dogs were presented with sets of one positive and two negative samples and were trained using positive reinforcement to hold their noses over the positive sample. The dogs performed moderately well in the final stage of training, with accuracy for Runnels of 0.817 and Cheaps of 0.647, both greater than the 0.333 expected by chance. During a double-blind detection test, dogs evaluated 123 unique and unfamiliar samples that were presented as 41 sets (3 samples per set), with both the dog handler and data recorder blinded to the positive sample location. Each dog was tested twice on each set of samples. Detection test accuracy was slightly better than chance for Cheaps at 0.451 (95% CI: 0.344-0.559) and was no better than chance for Runnels at 0.390 (95% CI: 0.285-0.496. Overall accuracy was 0.421 (95% CI: 0.345-0.496). When dogs' consensus response on each sample set was considered, accuracy was 0.537 (95% CI: 0.384-0.689). Detection accuracy also varied by sample lot. While dogs showed some ability to discriminate between BRD-affected and healthy cattle using nasal swabs, the complexity of this task suggests that more testing is needed before determining whether dogs could be effective as a screening method for BRD.

Impacts of commingling cattle from different sources on their physiological, health, and performance responses during feedlot receiving.

This experiment compared physiological, health, and performance responses of beef heifers assigned to different commingling schemes (one, two, or four sources per pen) during a 56-d feedlot receiving period. Ninety-six recently weaned Angus-influenced heifers were obtained from an auction facility. Heifers originated from four cow-calf ranches, and were reared in the same herd within each ranch since birth. Heifers were loaded into two livestock trailers at the auction yard (two sources per trailer; d -2), arranged in two sections of each trailer according to source, and transported for 10 h to stimulate the stress of a long-haul. Heifers were not mixed with cohorts from other sources prior to and at the auction yard. Upon arrival (d -2), shrunk body weight (BW) was recorded and heifers were maintained in four paddocks by source with ad libitum access to a complete starter feed and water for 36 h. On d 0, heifers were ranked by source and shrunk BW and allocated to 1 of 24 drylot pens (four heifers per pen) containing: 1) heifers from a single source (1SRC, n = 8), 2) heifers from two sources (2SRC, n = 8), or 3) heifers from four sources (4SRC, n = 8). From d 0 to d 55, heifers had free-choice access to the complete starter feed and water. Heifers were assessed daily for symptoms of bovine respiratory disease (BRD), and feed intake was recorded from each pen daily. Blood samples were collected on d 0, d 6, d 13, d 27, d 41, and d 55, and shrunk BW (after 16 h of water and feed withdrawal) was recorded on d 56 for average daily gain (ADG). No treatment differences were noted (P ≥ 0.56) for heifer ADG (mean ± SE = 0.853 ± 0.043 kg/d), final shrunk BW, feed intake, and feed efficiency. No treatment differences were noted (P ≥ 0.27) for plasma concentrations of cortisol and haptoglobin, and serum concentrations of antibodies against BRD viruses and Mannheimia haemolytica. No treatment differences were noted (P ≥ 0.17) for incidence of BRD (mean ± SE = 59.3 ± 5.0%) or mortality. The proportion of heifers diagnosed with BRD that required three antimicrobial treatments to regain health increased linearly (P = 0.03) according to the number of sources (0.0, 12.3, and 20.8% of 1SRC, 2SRC, and 4SRC heifers, respectively; SEM = 7.0). Hence, commingling heifers from different sources did not impact performance, physiological responses, and BRD incidence during a 56-d receiving period, although recurrence of BRD after the second antimicrobial treatment increased according to commingling level.

Administering an appeasing substance to beef calves at weaning to optimize productive and health responses during a 42-d preconditioning program.

This experiment evaluated the impacts of administering a bovine appeasing substance (BAS) to beef calves at weaning on their performance, physiological responses, and behavior during a 42-d preconditioning program. Eighty calves (40 heifers and 40 steers; 90% British × 10% Nellore) were weaned at 233 ± 2 d of age (day 0); ranked by sex, weaning age, and body weight (BW); and assigned to receive BAS (IRSEA Group, Quartier Salignan, France; n = 40) or placebo (diethylene glycol monoethyl ether; CON; n = 40). Treatments (5 mL) were topically applied to the nuchal skin area of each animal following dam separation. Within treatment, calves were allocated to one of eight drylot pens (four pens per treatment; pen being the experimental unit) and received a free-choice total mixed ration (TMR) from day 0 to 42, intake of which was assessed daily. Live behavior observations were conducted on days 1, 2, 4, 8, 16, and 32. Temperament was assessed and blood samples were collected via jugular venipuncture on days -21, 0, 3, 7, 14, 28, and 42. Hair samples were collected from the tail switch on days 0, 14, 28, and 42. Calves were vaccinated against bovine respiratory disease viruses on days -21 and 0. Average daily gain from day 0 to 42 did not differ between treatments (P = 0.57) but was greater (P = 0.05) in BAS vs. CON calves from day 0 to 28. Intake of TMR was greater (P = 0.05) during the first week for BAS vs. CON calves (treatment × week; P = 0.08). The mean proportion of calves feeding simultaneously and performance of social and play behaviors were greater (P ≤ 0.05) for BAS vs. CON calves. Escape attempts were greater (P < 0.01) for BAS vs. CON calves on day 1 (treatment × day; P = 0.03). Exit velocity was greater (P = 0.04) for CON vs. BAS calves on day 14 and tended (P = 0.10) to be greater for CON vs. BAS calves on day 7 (treatment × day; P = 0.03). Mean plasma concentrations of haptoglobin were greater (P = 0.02) in CON vs. BAS calves. Hair cortisol concentrations were greater (P = 0.05) in CON vs. BAS calves on day 14 (treatment × day; P = 0.03). Mean serum concentrations of antibodies against bovine viral diarrhea virus were greater (P = 0.02) in BAS vs. CON calves. Collectively, BAS administration to beef calves at weaning alleviated stress-induced physiological reactions, improved temperament evaluated via chute exit velocity, enhanced humoral immunity acquired from vaccination, and appeared to have accelerated adaptation to novel management scheme and environment.

A Scoping Review: The Impact of Housing Systems and Environmental Features on Beef Cattle Welfare.

Housing systems and environmental features can influence beef cattle welfare. To date, little information has been synthesized on this topic. The aim of this scoping review was to examine the relationship between housing and welfare status, so that beef cattle producers and animal scientists can make informed decisions regarding how their housing choices could impact beef cattle welfare. Housing features were categorized by floor type, space allowance and shade availability, as well as the inclusion of enrichment devices or ventilation features. Evaluation of space allowances across feedlot environments determined behavioral and production benefits when cattle were housed between 2.5 m2 to 3.0 m2 per animal. Over 19 different flooring types were investigated and across flooring types; straw flooring was viewed most favorably from a behavioral, production and hygiene standpoint. Veal calves experience enhanced welfare (e.g., improved behavioral, physiological, and performance metrics) when group housed. There is evidence that the implementation of progressive housing modifications (e.g., shade, environmental enrichment) could promote the behavioral welfare of feedlot cattle. This review presents the advantages and disadvantages of specific housing features on the welfare of beef cattle.

Cattle adapted to tropical and subtropical environments: social, nutritional, and carcass quality considerations.

Beef production needs to increase from 60 million to 130 million tons by 2050 to feed a growing world population, and 70% of this production increase is expected from beef industries located in subtropical and tropical regions of the world. Bos indicus-influenced cattle predominate in these regions but are often managed using practices developed for Bos taurus breeds reared in temperate climates. Hence, a fundamental step to meet the increasing global demand for beef is to develop specific management for B. indicus-influenced cattle in tropical or subtropical environments. Bos taurus and B. indicus are different subspecies, and diverge in social and biological functions due to selection pressure caused by complex evolutionary and domestication processes. Bos indicus cattle display different social responses compared with B. taurus counterparts, which must be taken into account by management planning as these traits directly impact cattle performance and welfare. In tropical and subtropical regions, warm-season perennial C4 grasses are the dominant forages, and their availability has a significant influence on the productivity of beef cattle systems. The resilience of C4 grasses under adverse conditions is one of their most important characteristics, even though these forages have reduced nutritive value compared with forages from temperate climates. Accordingly, nutritional planning in tropical and subtropical conditions must include management to optimize the quantity and quality of C4 forages. Nutritional requirements of cattle raised within these conditions also require special attention, including inherent metabolic compromises to cope with environmental constraints and altered energy requirements due to body composition and heat tolerance. Nutritional interventions to enhance beef production need to be specifically tailored and validated in B. indicus-influenced cattle. As an example, supplementation programs during gestation or early life to elicit fetal programming or metabolic imprinting effects, respectively, yield discrepant outcomes between subspecies. Bos indicus-influenced cattle produce carcasses with less marbling than B. taurus cattle, despite recent genetic and management advances. This outcome is mostly related to reduced intramuscular adipocyte volume in B. indicus breeds, suggesting a lesser need for energy stored intramuscularly as a mechanism to improve thermotolerance in tropical and subtropical climates.

Evaluating equine feeding behavior utilizing GrowSafe Systems: a pilot study.

Equine research and management is limited to single-housing systems if individual animal intake is to be precisely recorded. Even then, dry forage intake is difficult to quantify accurately due to stomping or mixing hay with fecal matter and bedding. In cattle management, GrowSafe Systems (GrowSafe) is a commonly used tool to closely monitor individual animal feeding data using radio frequency identification (RFID) tag technology. Animals are equipped with a unique RFID tag that is read by the feed bunks each time the animal lowers its head into the bunk to consume feed. The objectives of this pilot study were 1) to test the feasibility of use of the GrowSafe system with horses by measuring intake of dry hay and 2) to characterize feeding behaviors of horses in an individually housed (without competition) or group-housed (with competition) setting. To test the hypothesis that horses would consume more hay when individually (NOCOMP) compared to group-housed (COMP), 10 mature Quarter Horses (14 ± 1.5 yr) were placed in one of four pens containing GrowSafe feed bunks in a 4-wk crossover design consisting of two 2-wk treatment periods. Pen 1 contained five horses with access to two GrowSafe bunks (Period 1: n = 4 mares, n = 1 gelding; Period 2: n = 5 geldings); pens 2, 3, and 4 contained one horse each with access to one bunk. Horses were individually fed 0.25% body weight (BW; dry matter [DM] basis) of a commercial concentrate once per day and were allowed Coastal bermudagrass hay in the GrowSafe bunks ad libitum. Although five horses were used in the group-housed (COMP) pen to more closely mimic a true group environment, only data from horses that experienced both housing systems (n = 3 mares and n = 3 geldings) were used for statistical analyses. Hourly (P = 0.008) and daily (P = 0.003) durations of hay feeding were higher for NOCOMP compared to COMP horses, and total daily intake (g DM/kg BW) of NOCOMP horses tended to be greater (P = 0.09) than COMP horses. Conversely, eating rate (g DM/kg BW/min) was greater (P = 0.04) for COMP compared to NOCOMP mares but was unaffected by housing in geldings. The GrowSafe system may provide an opportunity for efficient and effective monitoring of individual horse feed intake and feeding behavior in group-housing situations in horses.

Erratum: Evaluating equine feeding behavior utilizing GrowSafe Systems: a pilot study.

[This corrects the article DOI: 10.1093/tas/txz002.].

Assessing Activity and Location of Individual Laying Hens in Large Groups Using Modern Technology.

Tracking individual animals within large groups is increasingly possible, offering an exciting opportunity to researchers. Whereas previously only relatively indistinguishable groups of individual animals could be observed and combined into pen level data, we can now focus on individual actors within these large groups and track their activities across time and space with minimal intervention and disturbance. The development is particularly relevant to the poultry industry as, due to a shift away from battery cages, flock sizes are increasingly becoming larger and environments more complex. Many efforts have been made to track individual bird behavior and activity in large groups using a variety of methodologies with variable success. Of the technologies in use, each has associated benefits and detriments, which can make the approach more or less suitable for certain environments and experiments. Within this article, we have divided several tracking systems that are currently available into two major categories (radio frequency identification and radio signal strength) and review the strengths and weaknesses of each, as well as environments or conditions for which they may be most suitable. We also describe related topics including types of analysis for the data and concerns with selecting focal birds.

Individual Consistency of Feather Pecking Behavior in Laying Hens: Once a Feather Pecker Always a Feather Pecker?

The pecking behavior [severe feather, gentle feather, and aggressive pecks (AP)] of individual White Shaver non-cage laying hens (n = 300) was examined at 21, 24, 27, 32, and 37 weeks. Hens were housed in 30 groups of 10 hens each and on 3 cm litter with access to a feeder, perch, and two nest boxes. The number of severe feather pecks given (SFPG) and received (SFPR) was used to categorize hens as feather peckers (P), victims (V), neutrals (N), or feather pecker-victims (PV) at each age. Hens categorized as PV exhibited pecking behaviors similar to P and received pecks similar to V. SFP given were correlated with APs given, but not with gentle feather pecks (GFP) given throughout the study. State-transition plot maps illustrated that 22.5% of P remained P, while 44% of PV remained PV throughout the duration of the study. Lifetime behavioral categories identified hens as a consistent feather pecker (5%), consistent neutral (3.9%), consistent victim (7.9%), consistent feather pecker-victim (29.4%), or inconsistent (53.8%) in their behavioral patterns throughout their life. Consistent feather peckers performed more SFP than hens of other categories, and consistent neutral hens received fewer GFP than consistent feather PV. No differences in corticosterone or whole blood serotonin levels were observed among the categories. Approximately, half of the population was classified as a feather pecker at least once during the study, while the remainder was never categorized as a feather pecker. Therefore, even if the development and cause of feather pecking may be multifactorial, once the behavior has been developed, some hens may persist in feather pecking. However, as some hens were observed to never receive or perform SFP, emphasis should be made to select for these hens in future breeding practices.

Moving GIS research indoors: spatiotemporal analysis of agricultural animals.

A proof of concept applying wildlife ecology techniques to animal welfare science in intensive agricultural environments was conducted using non-cage laying hens. Studies of wildlife ecology regularly use Geographic Information Systems (GIS) to assess wild animal movement and behavior within environments with relatively unlimited space and finite resources. However, rather than depicting landscapes, a GIS could be developed in animal production environments to provide insight into animal behavior as an indicator of animal welfare. We developed a GIS-based approach for studying agricultural animal behavior in an environment with finite space and unlimited resources. Concurrent data from wireless body-worn location tracking sensor and video-recording systems, which depicted spatially-explicit behavior of hens (135 hens/room) in two identical indoor enclosures, were collected. The spatial configuration of specific hen behaviors, variation in home range patterns, and variation in home range overlap show that individual hens respond to the same environment differently. Such information could catalyze management practice adjustments (e.g., modifying feeder design and/or location). Genetically-similar hens exhibited diverse behavioral and spatial patterns via a proof of concept approach enabling detailed examinations of individual non-cage laying hen behavior and welfare.

When continuous observations just won't do: developing accurate and efficient sampling strategies for the laying hen.

Continuous observation is the most accurate way to determine animals' actual time budget and can provide a 'gold standard' representation of resource use, behavior frequency, and duration. Continuous observation is useful for capturing behaviors that are of short duration or occur infrequently. However, collecting continuous data is labor intensive and time consuming, making multiple individual or long-term data collection difficult. Six non-cage laying hens were video recorded for 15 h and behavioral data collected every 2 s were compared with data collected using scan sampling intervals of 5, 10, 15, 30, and 60 min and subsamples of 2 second observations performed for 10 min every 30 min, 15 min every 1 h, 30 min every 1.5 h, and 15 min every 2 h. Three statistical approaches were used to provide a comprehensive analysis to examine the quality of the data obtained via different sampling methods. General linear mixed models identified how the time budget from the sampling techniques differed from continuous observation. Correlation analysis identified how strongly results from the sampling techniques were associated with those from continuous observation. Regression analysis identified how well the results from the sampling techniques were associated with those from continuous observation, changes in magnitude, and whether a sampling technique had bias. Static behaviors were well represented with scan and time sampling techniques, while dynamic behaviors were best represented with time sampling techniques. Methods for identifying an appropriate sampling strategy based upon the type of behavior of interest are outlined and results for non-caged laying hens are presented.