On-farm detection of claw lesions in dairy cows based on acoustic analyses and machine learning.

Claw lesions are a serious problem on dairy farms, affecting both the health and welfare of the cow. Automated detection of lameness with a practical, on-farm application would support the early detection and treatment of lame cows, potentially reducing the number and severity of claw lesions. Therefore, in this study, a method was proposed for the detection of claw lesions based on the acoustic analysis of a cow's gait. A panel was constructed to measure the impact sound of animals walking over it. The recorded impact sound was edited, and 640 sound files from 64 cows were analyzed. The classification of animal-lameness status was performed using a machine-learning process with a random forest algorithm. The gold standard was a 2-point scale of hoof-trimming results (healthy vs. affected), and 38 properties of the recorded sound files were used as influencing factors. A prediction model for classifying the cow lameness was built using a random forest algorithm. This was validated by comparing the reference output from hoof-trimming with the model output concerning the impact sound. Altering the likelihood settings and changing the cutoff value to predict lame animals improved the prediction model. At a cutoff at 0.4, a decreased false-negative rate was generated, and the false-positive rate only increased slightly. This model obtained a sensitivity of 0.81 and a specificity of 0.97. With this procedure, Cohen's Kappa value of 0.80 showed good agreement between model classification and diagnoses from hoof-trimming. In summary, the prediction model enabled the detection of cows with claw lesions. This study shows that lameness can be detected by machine learning from the impact sound of hoofs in dairy cows.

Histologically validated scoring system for the assessment of hock burn in broilers.

The assessment of bird-based welfare indicators plays an important role in the evaluation of bird welfare. The aim of the study was to histologically validate a visual scoring system for hock burn in broilers and to detect threshold values of a visual score to define welfare-relevant alterations in terms of mild lesions or ulcers of the hock. We collected 200 hocks of 39- to 42-day-old Ross 308 broilers after the slaughter process. Each hock was scored visually ("macro scores" 0-4) and evaluated histologically ("micro scores" 0-3), with high scores representing more severe lesions. Although we found a tendency for higher micro scores with increasing macro scores, an exact allocation of macro to micro scores was not possible. For example, macro score 1 could represent micro scores 1, 2 and 3, whereas macro scores 3 and 4 always represented micro score 3 (ulcer). The conditional probability of certain micro scores for given macro scores was estimated using a multinomial logistic regression model. Ulcer showed the highest probability at macro score 1, whereas mild lesions were not found to have an estimated highest probability at any macro score. The depth of inflammation of hock burn lesions increased with increasing macro scores up to macro score 3 with an average depth of 1019 µm. Visually more severe and deeper lesions were also histologically rated with higher scores. Thus, considering limitations, the herein validated macroscopic assessment scheme for hock burn allows an estimation of histological alterations in hocks of broilers.RESEARCH HIGHLIGHTS Histological validation of a visual assessment scheme for hock burn in broilers.Tendency for higher micro scores with increasing macro scores.Estimation of histological score via macro score possible with limitations.Histological depth of inflammation increased with an increasing macro score.

Inhibition of heat stress-related apoptosis of chicken myocardial cells through inducing Hsp90 expression by aspirin administration in vivo.

1. This experiment investigated the anti-apoptosis effects and the mechanism of aspirin action in the heat shock response of chicken myocardial cells in vivo, via changes in the heat stress (HS) protein Hsp90 and the rate of apoptosis. Broiler chickens were administered aspirin (1 mg/kg body weight) 2 h before exposure to HS, and then exposed to 40 ± 1°C for 0, 1, 2, 3, 5, 7, 10, 15 and 24 h. 2. The induction and consumption of the HS factor heat shock factor (HSF)-1, and reductions of HSF-2 and HSF-3 induced by HS led to a delay in Hsp90 expression. HSF-1, 2 and 3 regulation of hsp90 expression in turn inhibited the synthesis and activation of protein kinase ß (Akt), which resulted in a significant increase in caspase-3 at 2 and 10 h, caspase-9 from 1 to 7 h (except at 5 h), and the heat-stressed apoptosis of the myocardial cells. 3. Administration of aspirin changed the expression patterns of HSF-1, 2 and 3 such that the expression of Hsp90 protein was significantly upregulated (by 2.3-4.1 times compared with that of the non-treated cells). The resultant increase in Akt expression and activation, compared with the HS group, inhibited caspase-3 and caspase-9 activities and reduced the myocardial cells apoptosis rate (by 2.14-2.56 times). 4. Aspirin administration could inhibit heat-stressed apoptosis of myocardial cells in vivo and may be closely associated with its promotion of HS response of chicken hearts, especially Hsp90 expression.

MicroRNA expression profiling of porcine mammary epithelial cells after challenge with Escherichia coli in vitro.

BACKGROUND: Coliform mastitis is a symptom of postpartum dysgalactia syndrome (PDS), a multifactorial infectious disease of sows. Our previous study showed gene expression profile change after bacterial challenge of porcine mammary epithelial cells (PMECs). These mRNA expression changes may be regulated through microRNAs (miRNAs) which play critical roles in biological processes. Therefore, miRNA expression profile was investigated in PMECs. RESULTS: PMECs were isolated from three lactating sows and challenged with heat-inactivated potential mastitis-causing pathogen Escherichia coli (E. coli) for 3 h and 24 h, in vitro. At 3 h post-challenge with E. coli, target gene prediction identified a critical role of miRNAs in regulation of host immune responses and homeostasis of PMECs mediated by affecting pathways including cytokine binding (miR-202, miR-3277, miR-4903); IL-10/PPAR signaling (miR-3277, miR-4317, miR-548); and NF-ĸB/TNFR2 signaling (miR-202, miR-2262, miR-885-3p). Target genes of miRNAs in PMECs at 24 h were significantly enriched in pathways associated with interferon signaling (miR-210, miR-23a, miR-1736) and protein ubiquitination (miR-125, miR-128, miR-1280). CONCLUSIONS: This study provides first large-scale miRNA expression profiles and their predicted target genes in PMECs after contact with a potential mastitis-causing E. coli strain. Both, highly conserved miRNAs known from other species as well as novel miRNAs were identified in PMECs, representing candidate predictive biomarkers for PDS. Time-dependent pathogen clearance suggests an important role of PMECs in inflammatory response of the first cellular barrier of the porcine mammary gland.

Determination of body width in brown and white layer pullets by image analyses.

1. Specific legal requirements for keeping pullets are not available in the European Union. However, two of the most important rearing factors for pullets are sufficient perching and feeder space. Both factors represent horizontal space dimensions which derive from the body width of the birds. 2. The body width of two strains of layer pullets (brown (BL) and white (WL) layer pullets) based on the measurement of distances in digital images was conducted on front-view digital photographs of BL and WL pullets taken at 8, 12 and 19 weeks of life. 3. Depending on live weight, age and body position, BL pullets measured an average body width between 10.70 ± 1.10 and 13.96 ± 1.11 cm. The width of WL pullets ranged from 10.30 ± 0.86 to 13.00 ± 1.14 cm. 4. Compared with WL, BL pullets occupied more horizontal space during rearing. Age influenced the body width of BL and WL pullets at the end of rearing. The tested body positions of the pullets did not affect the measured body width. 5. The biometric data obtained in this study are a useful basis for developing legal requirements for pullets, especially for defining minimum perch width and feeder space allowances.

Spatial requirements of poultry assessed by using a colour-contrast method (KobaPlan).

Sufficient floor space is a fundamental precondition for poultry to perform normal behavioural patterns. To calculate and determine stocking densities, it is essential to know the absolute minimum surface area required by any given animal (body space). Additional space is required for characteristic behaviours (behavioural space) and for adequate inter-individual distances, group sizes and room to perform social interactions have to be taken into account. To calculate body space, planimetric measurements were carried out by the colour contrast method "KobaPlan" in various poultry species in standing and sitting positions and at a number of different ages. They included laying hens (Lohmann brown (LB), Lohmann selected Leghorn (LSL)), broiler breeders (Ross, both genders), broiler chickens (Ross 308, both genders), turkeys (BUT 6, males), Peking ducks (Cherry Valley, both genders) and Muscovy ducks (Canedins R51, males). Depending on live weight, age, plumage condition and body position, LB hens occupied an average area between 401 cm(2) and 542 cm(2), LSL hens between 353 cm(2) and 445 cm(2), broiler breeder females between 440 cm(2) and 537 cm(2), broiler breeder males 623 cm(2) up to 945 cm(2), broiler chickens up to 434 cm(2), male fattening turkeys up to 1808 cm(2), Muscovy drakes up to 873 cm(2) and Peking ducks up to 627 cm(2). The values can be regarded as necessary minimum spatial requirements for the measured poultry species and genotype. The current method offers the potential to record the area occupied by animals exhibiting species-specific behavioural patterns.

HSP90 gene expression induced by aspirin is associated with damage remission in a chicken myocardial cell culture exposed to heat stress.

To understand the potential protection of heat shock protein 90 (HSP90) induced by aspirin against heat stress damage in chicken myocardial cells, enzyme activities related to stress damage, cytopathological changes, the expression and distribution of HSP90, and HSP90 mRNA levels in the myocardial cells exposed to heat stress (42°C) for different durations with or without aspirin administration (1 mg/ml, 2 h prior) in vitro were investigated. Significant increase of enzyme levels in the supernatant of heat-stressed myocardial cells and cellular lesions characterised by acute degeneration, karyopyknosis and karyorrhexis were observed, compared to non-treated cells. However, the lesions of cells treated with aspirin were milder, characterised by earlier recovery of enzyme levels to the control levels and no obvious heat stress-related cellular necrosis. Stronger positive signals in the cytoplasm and longer retention of HSP90 signal in nuclei were observed in aspirin-treated myocardial cells than those of only heat-stressed cells. HSP90 level in the aspirin-treated myocardial cells was 11.1-fold higher than that in non-treated cells, and remained at a high level at the early stage of heat stress, whereas it was just 4.1-fold higher in only heat-stressed cells and returned rapidly to a low level. Overexpression of HSP90 mRNA in aspirin-treated cells was observed throughout the experiment, whereas HSP90 mRNA decreased significantly only in heat-stressed cells. The early higher HSP90 expression induced by aspirin during heat stress was accompanied by decreased heat stress damage, suggesting that aspirin might play an important role in preventing myocardial cells from heat stress damage in vitro.

Association of heat shock protein 70 expression with rat myocardial cell damage during heat stress in vitro and in vivo.

To investigate the mechanism of sudden death as a result of stress-induced damage to heart tissue and myocardial cells and to investigate the cardioprotective role of Hsp70 during heat stress, the distribution and expression of Hsp70 was evaluated in the heart cells of heat-stressed rats in vivo and heat-stressed H9c2 cells in vitro. After exposure to heat stress at 42°C for different durations, we observed a significant induction of CK, CK-MB, and LDH as well as pathologic lesions characterized by acute degeneration, suggesting that cell damage occurs from the onset of heat stress. Immunocytochemistry showed that Hsp70 was distributed mainly in the cytoplasm of myocardial cells in vivo and in vitro. Hsp70-positive signals in the cytoplasm were more prominent in intact areas than in degenerated areas after 60 min of heat stress. Hsp70 protein levels in myocardial cells in vitro decreased from the beginning to the end of heat stress. Hsp70 protein levels in rat heart tissues in vivo decreased gradually with prolonged heat stress, with a slight increase at the beginning of heat stress. These results indicate that Hsp70 plays a role in the response of cardiac cells to heat stress and that decreased Hsp70 levels are associated with damage to rat myocardial cells in vitro and in vivo. Significant differences were found in hsp70 mRNA, which began to increase after 20 min of heat stress in vitro and after 40 min in vivo. This indicates that hysteresis is involved in mRNA expression after heat stress in vivo.

Evaluation of Hsp47 expression in heat-stressed rat myocardial cells in vitro and in vivo.

The aim of the present study was to identify the correlation between expression of heat shock protein 47 (Hsp47) and stress injury in heat-stressed myocardial cells and to compare variations in Hsp47 expression in rat myocardial cells exposed to different heat stress for varying periods in vitro and in vivo. Exposure to heat stress at 42°C resulted in similar induction patterns of the heart damage-related enzyme aspartate aminotransferase in the supernatants of H9c2 cells and in the serum of rats. Histological analysis revealed that both H9c2 cells and heart tissues displayed cellular degeneration in response to different periods of heat stress. Hsp47 was constitutively expressed in the cytoplasm of H9c2 cells at all time points during heat stress, which was consistent with observations in heart fibers in vivo. Immunoblotting analysis revealed no significant difference between the expression of Hsp47 in H9c2 cells and heart tissue. However, the expression of hsp47 mRNA in response to heat stress was significantly increased in H9c2 cells at 60 min (P < 0.01) and 100 min (P < 0.01), which was comparable to that at 100 min (P < 0.01) in the rat heart. Thus, Hsp47 was elevated significantly after hyperthermia at the mRNA level but not at the protein level both in vitro and in vivo. The results suggest that Hsp47 turnover may increase during heat stress or that Hsp47 consumption exceeds its production.

Association of Hsp60 expression with damage to rat myocardial cells exposed to heat stress in vivo and in vitro.

To investigate the protective role of Hsp60 against stress damage and its role in the sudden death of stressed animals, changes in the levels of Hsp60 protein and hsp60 mRNA of myocardial cells in vivo and in vitro were studied. In addition, the relationship between Hsp60 expression and heat-induced damage was also studied. Rats were exposed to a temperature of 42° ± 1°C for 0, 20, 40, 60, 80, or 100 min. More than 50% of the rats died suddenly within 100 min. With increasing heat stress duration, hsp60 mRNA levels significantly increased in both in vivo and in vitro rat myocardial cells; however, a similar trend was not observed for Hsp60 protein levels. Although the changes observed in Hsp60 expression in myocardial cells in vitro were inconsistent with those of rat heart tissues in vivo, Hsp60 expression levels were consistent with the histopathological damage observed in myocardial cells both in vivo and in vitro. Differences in Hsp60 expression may reflect the degree of injury sustained by myocardial cells in vivo and in vitro. As a mitochondrial protein, Hsp60 represents a potential biomarker of heat stress, and may protect against heat stress induced myocardial cellular damage both in vivo and in vitro.

Use of an elevated platform with perforated surface and manure belt by fast-growing broilers on commercial farms.

Like other members of the species Gallus gallus, fast-growing broilers are motivated to perch. However, broilers in the European Union are kept in unstructured barns, with no opportunity to sit elevated and rest undisturbed. A possible solution to this problem is elevated platforms. The aim of this study was to evaluate the use of an elevated platform with perforated surface and manure belt by fast-growing broilers. On 2 commercial farms, an elevated platform was installed in 1 barn per farm. Approximately 35,000 Ross 308 broilers were housed in each barn for 3 fattening periods. On 1 d per wk, the number of broilers per m² on the platform and the ramp was determined every 30 min from video recordings. Besides, focal animals were observed to analyze their behavior on the platform at different ages and during light and dark periods. Broilers used the elevated platform and the ramp from the first week until the end of the fattening period (platform: 9.92 broilers per m², ramp: 6.47 broilers per m²), with a peak in the fourth week of life (platform: 13.00 broilers per m²). In wk 2, 4, and 5, platform use was higher during the light period than during the dark period. Broilers stayed longer on the platform in the dark period (dark: 01:54:23 [hh:mm:ss], light: 00:19:54 [hh:mm:ss]). In every phase of the fattening period, broilers on the platform were inactive to a high proportion (on average 80.60%). This indicates that broilers used the platform also for resting behavior. Thus, the elevated platform with perforated surface and manure belt is a suitable option to structure broiler barns. It allows the broilers to sit elevated and provides additional space. Factors such as a shallow ramp incline of 20°, wide ramps, and appropriate material used for the surface and ramps may have contributed to its high use by broilers of all ages. Further research is needed to evaluate the design of platforms that allow broilers to rest undisturbed.

Exploring the impact of biosecurity measures on somatic cell score in mountain dairy farms considering the CLASSYFARM system.

Biosecurity plays a crucial role in preventing the introduction of infectious diseases to a herd as well as the spread of diseases within or between animals and herds. In particular, biosecurity measures are crucial for maintaining animal health and reducing the need for the application of antibiotic substances for fighting the rising antibiotic resistance. The object of this cross-sectional study was to investigate the presence of biosecurity measures and their association with milk quality parameters, with a special focus on somatic cell count (SCC) - an indicator for udder health -, in small-scale mountain dairy farms. Therefore, for the very first time, the CLASSYFARM system was considered, which is a computer platform integrated into the Italian national veterinary portal, that processes a significant amount of data from various sources collected in the field or from other information systems (e.g. animal welfare, health status, biosecurity, antimicrobial use, slaughterhouse information). A total of 169 dairy farms were included in the study. Biosecurity measures, based on 15 questions required in the CLASSYFARM welfare assessment protocol, as well as information about husbandry systems, milking systems and pasture practices were gathered and combined with milk yield data, provided by the South Tyrolean dairy association. Farms only scored 44.00 points on average in a scoring system from 0.00 to 100.00 points that was be able to summarize 15 different biosecurity measures in one index. Our results show a clear negative correlation (-0.713) between the biosecurity index and somatic cell score (SCS) indicating that a higher level of biosecurity, which reflects the presence of biosecurity measures within a farm, is associated with lower SCC levels. Furthermore, we found significant correlations between SCS and milk production (-0.629), confirming that udder health is linked to higher milk production. Fat, protein, and the fat-to-protein ratio showed a positive correlation with SCS (0.281, 0.146, 0.106), likely to be caused by a concentration shift effect (dilution effect). Husbandry system, breed, milking system, and pasture practices seem to have an impact as well, but the main factor was the biosecurity score. This study highlights the importance of implementing biosecurity measures for ensuring animal health and thus productivity and quality in milk production, even in small-scale farms, which are characterized by limited structure availability and smaller herds compared to big dairy enterprises in the lowlands.

Occurrence of ultrasonographic assessed placental abnormalities, treatments, pregnancy outcome, and subsequent fertility on a large warmblood stud farm: A retrospective field study.

Little is known about the incidence and outcome of high-risk pregnancies in equine practice and clinical studies on spontaneous occurring placentitis cases and treatments are missing. Therefore, the aims of this retrospective field study were to (1) describe the incidence and severity of ultrasonographic assessed placental abnormalities (UPA) in 4,192 pregnancies on a large commercial warmblood stud farm in 2017 - 2019 and (2) characterize these UPA cases and their pregnancy outcome. UPA severity (Placental abnormality score (PSc) 1-3; low to high), nine treatment regimens (TM1-9) used in UPA cases and treatment duration as well as subsequent fertility were analyzed in the group of UPA mares. The proportion of pregnancies affected by UPA was 4.2 % (n=177/4192). Placental abnormality severity was scored as PSc1 (51.4 %), PSc2 (32.8 %) and PSc3 (15.8 %). The generalized mixed model revealed PSc was affected by mare age and mare status (own pregnancy (OP) or embryo transfer recipient (ER)) (P=0.035) with ER mares having increased PSc compared with mares having their own pregnancy. Abortion occurred in 17/177 (9.6 %) UPA pregnancies. Overall, at the end of the next season, 61.1 % of UPA mares were pregnant, 32.0 % barren, and 6.9 % open (n=175). Pregnancy was established in 62/91(68.1 %) of mares with PSc1, 31/58 (53.4 %) with PSc2 and 14/26 (53.8 %) with PSc3. Most pregnancies were achieved in the first 81/107 (75.7 %) or second 18/107 (16.8 %) inseminated cycle. In conclusion, early detection and treatment of ultrasonographic assessed placental abnormalities can save high-risk pregnancies in > 90 % of cases with a satisfying subsequent fertility.

Field trial: disinfection of contaminated anesthetic masks for piglets.

This paper aimed to assess the success of cleaning and disinfection on microbiological contamination of anesthetic masks, which were used for automated isoflurane anesthesia for surgical castration of male piglets. Data collection took place on 11 farms in Southern Germany between September 2020 and June 2022. Each farm was visited three times (one farm having two different anesthesia devices was visited six times), and microbiological assessments took place at four sample points (SP): after unpacking the masks (SP0), after disinfection before anesthesia (SP1), after anesthesia of all piglets to be castrated in this run (SP2), and after disinfection after anesthesia (SP3). The microbiological assessment included the determination of total bacteria count, total count of hemolytic and non-hemolytic mesophilic aerotolerant bacteria and a qualitative detection of indicator bacteria Escherichia (E.) coli, extended-spectrum beta-lactamase-producing E. coli (ESBL) and methicillin-resistant Staphylococcus aureus (MRSA). For analysis, a generalized linear mixed model was applied using farms and farm visits as random effects and sampling points nested in farm visits as fixed effect. The fixed effect was highly significant for all three variables (total bacteria count, total count of hemolytic and non-hemolytic mesophilic aerotolerant bacteria) (p < 0.001). The bacterial counts at SP0 were about the same as at SP3. Concerning indicator bacteria, their presence was highest at SP2 and lowest at SP3. No indicator bacteria were present at SP1. It can be concluded that disinfection of anesthetic masks, especially before performing anesthesia, may effectively protect piglets of the following batch against unwanted transmission of pathogens. These findings will help farmers plan cleaning and disinfection activities.

Estimation of variance components for postpartum dysgalactia syndrome in sows.

The postpartum dysgalactia syndrome (PDS) represents one of the most important diseases after parturition in sows. The genetic background of the disease has been investigated some time ago and heritability estimates around 0.10 have been obtained. To compute current estimates, a dataset of 1680 sampled sows and their 2001 clinically examined litters was used for variance components estimation with a threshold liability model. Affected sows were defined through clinical examination 12-48 h after parturition. Posterior mean of additive genetic variance was 0.10 and estimated heritability for PDS averaged 0.0879 with a 95% confidence interval of 0.0876 and 0.0881. The results are in agreement with those of other studies and emphasize the importance of considering the genetic predisposition for susceptibility to PDS as well as of additional factors including hygiene and management conditions.

Genetic aspects for the behaviour of lactating sows towards humans.

With the keeping of lactating sows in loose housing systems, ensuring work safety for stockpersons is gaining importance. Aim of the present study was to develop tests characterising the behaviour of lactating sows in farrowing environments with more freedom to move. The behaviour towards humans in different management procedures was examined. Emphasis was given to integrate tests into daily routines. The study was conducted in a nucleus herd with 771 purebred Landrace sows. Data were collected from October 2016 until December 2018. Sows were kept in individual indoor pens with movable farrowing crates in which the animals were restrained from 7 days antepartum (ap) to an average of 7 days postpartum (pp). The Dummy Arm Test (DAT; 1444 observations) was used to assess the sows' reaction towards a stockperson handling the piglets around day 4 pp (closed crates). With the Towel Test (TT; 2846 observations), the reaction of sows to a novel object and an unexpected situation was assessed. The Trough Cleaning Test (TCT; 2805 observations) described the sows' response to common procedures such as trough cleaning. TT and TCT were conducted on days 3 pp (closed crates) and 10 pp (open crates). Variance components of behavioural traits were estimated univariately with a linear animal model, and genetic correlations between traits were derived using a multivariate animal model in ASreml 3.0. Most sows showed no or only a slight reaction to human interactions without attempting to attack them. However, a strong defensive reaction of sows was recorded in 4.0% (TCT), 4.5% (TT), and 10.7% (DAT) of observations. This behaviour of sows was observed more frequently in the open than in the closed pen system. Estimates of heritabilities (h2 ± SE) were h2 = 0.17 ± 0.05 for behaviour of sows towards humans (DAT), h2 = 0.19 ± 0.04 for response of sows towards unexpected situations (TT), and h2 = 0.13 ± 0.04 for reactions of animals to TCT. Genetic correlations (rg ± SE) ranged from rg = 0.59 ± 0.37 between TT and TCT to rg = 0.77 ± 0.30 between TT and DAT. Our results show that the developed tests are suitable for assessing the behaviour of sows towards humans. Behavioural traits derived from these tests could be used as new phenotypes for the genetic selection of gentle and easy-to-handle sows. The genetic correlations of all tests studied were positive indicating related reaction patterns.

Effect of pens with an elevated platform on space utilization, skin lesions and growth performance in nursery pigs.

Environmental enrichment, adequate space and the ability to separate a living area into suitable functional areas are key elements for pig welfare. In this study, a two-level pen for nursery pigs was explored with the aim of analysing the use of space and the impact on pig behaviour, health and performance. Therefore, per batch, three experimental groups in pens with elevated platforms (two-level pens) and one control group in a standard pen were formed after weaning at the age of 4 weeks. Thereafter, groups were studied for a period of 6 weeks. In a total of eight batches (n = 882 pigs), the occurrence of skin lesions was determined per individual using a lesion score, and in seven batches (n = 761 pigs) individual daily weight gain was analysed. In five batches (n = 450 pigs), the individual use of space and the behaviour of pigs were investigated by direct observation (4 h per day on 2 days in weeks 1, 3 and 6 of the housing period, respectively). Results revealed that 98.9% of pigs (n = 445) were observed on the elevated platform at least once. The probability that an individual pig used the platform was significantly higher than 0.95 (P < 0.0001, confidence interval = 0.977, 1). The use of the platform was not affected by sex (odds ratio (OR) = 1.013, P = 0.937). However, the probability of observing an individual on the elevated platform increased with increasing BW during the experimental period (OR = 1.043, P < 0.0001). The presence of a platform decreased the probability of seeing a pig fighting (P = 0.014) and increased the probability of observing locomotor behaviour as opposed to lying (P < 0.0001). In two-level pens, pigs sustained fewer skin injuries than in standard pens (day 41: OR = 0.731, P < 0.0001). Over 41 days of experiment, pigs in two-level pens had higher daily weight gains than animals in standard pens (416 versus 393 g/day, P = 0.006). We conclude that usable space for nursery pigs can be effectively extended by introducing elevated platforms into the pigs' pen. Furthermore, pigs may benefit from two-level systems by establishing spaces for activity as well as for retreat from other pen mates, thereby reducing aggressive behaviour and social stress. The installation of two-level pens is therefore encouraged, if they are intended to provide more space than legally stipulated.

Short communication: Evaluation of the sample size of individual indicators in gestating sows concerning the Welfare Quality® protocol applied to sows and piglets.

Sample sizes of welfare assessment protocols must warrant to reflect prevalences on-farm properly - regardless of farm size. Still, solely a fixed sample size was specified for the Welfare Quality® protocol for sows and piglets. The present study investigated whether animals may be assessed from only one body side as applied in the protocol and whether the pre-set sample size of 30 animals mirrors the prevalences of the animal-based indicators on-farm in the gestation unit considering different farm sizes. All indicators were assessed for both sides of an animal's body by one observer on 13 farms in Germany, which were visited five times within 10 months. The farm visits were treated as independent since different animals were housed in the gestation units. The number of sows in the gestation units varied between 18 and 549 animals. The comparison of sides was carried out calculating exact agreement between animals' sides and a Wilcoxon signed-rank test (W). The results signified that it is sufficient to assess the animal from one side (exact agreement: 88.3% to 99.5%, except for bursitis (70.0%); W: P-values 0.14 to 0.92). However, if side preferences existed in the indicator bursitis a potential bias must be considered. In the following, the sample size was evaluated by comparing samples' prevalences against true prevalence, that is, the prevalence of all observed animals in the gestation unit in each farm visit. Therefore, subsets of data were generated by applying simple random sampling without replacement. The samples randomly included the animals' right or left sides. Linear regression was rated as appropriate provided: coefficient of determination R2 ≥ 0.90, slope = 1 and intercept = 0 signifying exact agreement. The results revealed that the sample size required by the protocol and the application of calculation formulas are solely appropriate to mirror the prevalences of frequent indicators in the gestation unit, for example, bursitis (mean prevalence 34.4%). Using a proportion of animals, for example, a sample of 30% of all observed animals in a farm visit, pointed out that proportions must increase with indicators' underlying prevalence narrowing 0.00%. Local infections (mean prevalence 13.3%) needed samples including 60% of all observed animals in each farm visit, whereas vulva lesions (mean prevalence 7.28%) only reached accuracy with the inclusion of 70% of the animals. Indicators with a mean prevalence of <1% were not analysed but can most likely only be ascertained by the assessment of all animals.

Corticosterone in feathers of laying hens: an assay validation for evidence-based assessment of animal welfare.

Studies indicate that the evaluation of animal welfare in birds may be carried out with the measurement of the stress-related hormone corticosterone in feathers. However a standardized procedure for corticosterone measurements in feathers is lacking, a validation needs to be carried out for each new species before implementation. The aim of the present study was to establish a valid method to measure corticosterone concentrations in feathers of laying hens in a precise and repeatable manner using an established and commercially available ELISA. Validation was performed with feather pools of tail and interscapular feathers of commercial Lohmann Brown laying hens. Assessment groups, consisting of 5 replicates, were created. All replicates of an assessment group were processed at the same time. Each replicate was run in 4 repetitions by ELISA. Intra-assay and interassay CV was 7.5 and 6.4%, respectively. The serial dilution showed linearity and parallelism. Examining the hormone extraction efficiency by using different methanol volumes resulted in no statistical differences (P > 0.05). Pulverized feathers showed higher corticosterone values than minced feathers (P > 0.05). Differences were shown between 2 feather types (tail vs. interscapular feathers; P < 0.05), as well as between vane and rachis (P < 0.05). Performance of a freeze-thaw cycle led to a decrease of corticosterone concentrations in the samples. A possible effect of UV-A radiation on the stability of corticosterone in the feathers was not found (P > 0.05). With the present study, a valid protocol, feasible for analyzing feather pools of laying hens, was developed. It may provide fundamentals for further investigations on corticosterone in feathers as a noninvasive indicator to evaluate aspects of animal welfare.

Effects of a partially perforated flooring system on animal-based welfare indicators in broiler housing.

Alternative flooring designs in broiler housing have been the subject of intensive research. Research comparing different floor types with a focus on animal-based welfare indicators might be of special interest to meet the animal's needs. This case-control study investigated the effect of a partially perforated vs. a littered flooring system on health- and behavior-based welfare indicators of fast-growing Ross 308 broilers. Furthermore, production performance was assessed. The experimental barn was partially (50%) equipped with a perforated floor directly underneath the feeders and water lines accessible by perforated ramps. Conventional wood shavings were used in the control barn, as usual in practice. There were 4 fattening periods (repetitions) of 31 to 32 D performed with 500 animals per barn (final density of 39 kg m-2). Beside the flooring system, management conditions were identical. Health- and behavior-based welfare indicators were assessed weekly. Production performance indicators were measured continuously during animal control. During the avoidance distance test, animals were less fearful on day 21 (P = 0.010) and tended to be less fearful on day 28 (P = 0.083) in the barn with the partially perforated flooring system compared to the littered control barn. More animals around the novel object were also assessed in the barn with the partially perforated flooring system during the novel object test on day 1 (P < 0.001) and a tendency was found on day 28 (P = 0.064). Results showed that the partially perforated flooring system had a positive influence on foot pad dermatitis from day 14 (all P ≤ 0.007) and hock burn on day 28 (P < 0.001). With regard to the production performance, animals showed no differences in final body weight for both floor types. In this study, the partially perforated flooring system had a positive effect on animal health and behavior as indicated by welfare indicators without a reduction in production performance.