Validation and improvement of a multiplex PCR method to detect murine Helicobacter species in feces samples of mice.

BACKGROUND: Murine Helicobacter species have gained increasing awareness in mouse facilities over the last years. Infections with Helicobacter species may have an impact effect on the health of mice and might pose a zoonotic risk to researchers. To minimize the interference with experiments and hence contribute to the 3Rs, a reliable method of monitoring Helicobacter infections in animal facilities needs to be available. The aim of this study was to improve and validate the detection of the most common murine Helicobacter species. MATERIAL AND METHODS: A multiplex PCR assay was developed for identification of Helicobacter hepaticus, H. bilis, H. muridarum, H. rodentium, and H. typhlonius that could simultaneously detect these five strains in fecal samples. To ensure the quality of the results, the method was validated based on recommendations for in-house developed tests. RESULTS: The method established was highly sensitive and specific. All five strains were detectable with a detection limit of 102 bacteria. Eight different mouse facilities were tested with the validated assay, and the following prevalence were found: H. rodentium 57%, H. hepaticus 46%, H. typhlonius 17%, H. bilis 12%, and H. muridarum 0%. CONCLUSION: The multiplex PCR is a reliable, economic, and time-saving diagnostic tool for routine health monitoring. Further prevalence studies are needed to confirm the high prevalence and hence importance of H. rodentium, as until now this agent is not yet listed in FELASA recommendations.

New insights into the association between lameness, behavior, and performance in Simmental cows.

In a herd of 100 milking Simmental cows, data of performance and behavior parameters were collected automatically with different systems such as pedometers, an automatic milking system, and automatic weighing troughs for 1 yr. Performance measures were several milking-related parameters, live weight, as well as feed intake. Behavior-associated measures were feeding behavior (e.g. feeding duration, number of visits to the trough, and feeding pace) as well as activity such as lying duration, number of lying bouts, and overall activity. In the same time, lameness status of every cow was assessed with weekly locomotion scoring. According to the score animals were then classified lame (score 4 or 5) or nonlame (score 1, 2, or 3). From these data in total, 25 parameters summarized to daily values were evaluated for their ability to determine the lameness status of a cow. Data were analyzed with a regularized regression method called elastic net with the outcome lame or nonlame. The final model had a high prediction accuracy with an area under the curve of 0.91 [95% confidence interval (CI) = 0.88-0.94]. Specificity was 0.81 (95% CI = 0.73-0.85) and sensitivity was 0.94 (95% CI = 0.88-1.00). The most important factors associated with a cow being lame were number of meals, average feed intake per meal, and average duration of a meal. Lame cows fed in fewer and shorter meals with a decreased intake per meal. Milk yield and lying-behavior-associated parameters were relevant in the model, too, but only as parts of interaction terms demonstrating their strong dependence on other factors. A higher milk yield only resulted in higher risk of being lame if feed intake was decreased. The same accounts for lying duration: only if lying time was below the 50% quantile did an increased milk yield result in a higher risk of being lame. The association of lameness and daily lying duration was influenced by daily feeding duration and feeding duration at daytime. The results of the study give deeper insights on how the association between behavior and performance parameters and lameness is influenced by intrinsic factors in particular and that many of these have to be considered when trying to predict lameness based on such data. The findings lead to a better understanding why, for instance, lying duration or milk yield seem to be highly correlated with lameness in cows but still have not been overly useful as parameters in other lameness detection models.

Movements after Captive Bolt Stunning in Cattle and Possible Animal- and Process-Related Impact Factors-A Field Study.

Movements in cattle after captive bolt stunning cause problems in the slaughter process and lead to uncertainties in assessing stunning effectiveness. The objective of this study was to categorize and quantify these movements and determine animal- and process-related impact factors, as well as connections to stunning effectiveness and shooting position. In total 2911 cows, heifers, and bulls (dairy, beef, and crossbreeds) were examined (mean age 3.02 years). Movements from landing until at least four minutes after sticking were recorded by action cams (Apeman® A100). Nine movement categories were defined ("kicking hind limb", "twitching", "bending and stretching hind limb", "lifting and bending forelimb", "body arching laterally", "body arching ventrally", and "arching backwards"). According to the movement severity, a score was assigned to each category. The scores were summed, either for certain process intervals, e.g., LANDING (ejection from the stunning box), HOISTING, or STICKING, or for the total time between LANDING and end of the FOURTH MINUTE OF BLEEDING (sum score). Statistical analysis (ANOVA) was performed on the scores. Only 6.6% of cattle showed no movement. Most movements occurred during STICKING and FIRST MINUTE OF BLEEDING, occurring rarely up to 8 min after sticking. While cows moved most at LANDING, bulls and heifers moved more if all process intervals were considered. The sum score was highest in German Angus, Charolais, and Limousin and lowest in Brown Swiss and Simmental. The score at LANDING was highest in German Angus and Black Holstein. The use of pneumatic stunners and an increase in bolt-exit length significantly reduced movements. No impact of stunning effectiveness on movements was found, but only 19 cattle showed reduced effectiveness.

Behavioral and hormonal assessment of stress in foals (Equus caballus) throughout the weaning process.

This study had the aim to demonstrate the midterm effects (three weeks) of weaning on foals' welfare. For this purpose, foals' behavioral changes and fecal levels metabolites of cortisol were evaluated. The observations took place at the state stud farm of Baden-Wuerttemberg in Germany. Ten foals (six colts and four fillies) were observed from one day before weaning up until three weeks after weaning. Weaning was divided into three blocks, the first in September, the second in October, and the last in November. The behavioral observation was done during an eight-hour period between 7:00 a.m. and 5:00 p.m. The observer documented the exact behavior shown by the foal every five minutes during the eight hours. To scale the stress experienced by the foal, the glucocorticoid metabolite 11,17-dioxoandrostane was measured with the 11-oxoetiocholanolone enzyme immunoassay, which allows assessing the foal's plasma cortisol level changes throughout the trail through fecal samples. All foals displayed a distinct hormonal stress response to the weaning process through increased fecal cortisol metabolite levels. Their body posture distribution took a shift from mainly moving before weaning to mainly standing during the three weeks after weaning. Compared with the day before weaning, the foals showed less active behavior and significantly increased their resting behavior. Regarding the overall resting behavior, the weaned foals initially increased their time spent resting in a lying position during daytime and then started to decrease the time lying. After weaning, the foals showed a significant increase in resting while standing. In conclusion, the foals showed an expected behavioral development and an expected curve of cortisol metabolite values throughout the study. However, it seemed that the changes had not returned "back to normal" at three weeks after weaning. Therefore, we suggest that weaned foals need a minimum of three weeks to acclimate to the new situation.

Behavior of broilers and impacts occurring to them during mechanical loading under field conditions.

Mechanical loading has been used increasingly in recent years alongside the manual loading of broilers. The aim of this study was to analyze the influence of various factors on the behavior of broilers and the impacts occurring to broilers during loading with a loading machine in order to identify risk factors and thus improve animal welfare. During 32 loadings we assessed escape behavior, wing flapping, flip, bumping into an animal, and bumping against the machine or container by evaluation of video recordings. The parameters were analyzed for influences of rotation speed, container type (GP container vs. SmartStack container), husbandry system ("Indoor Plus" vs. "Outdoor Climate"), and season. In addition, the behavior and impact parameters were correlated with the loading-related injuries. The results showed that an increase in rotation and conveyor belt speed was associated with an elevated risk of all behaviors or impacts, except for a lower risk of escape behavior. The seasonal influence showed that the risk of wing flapping, bumping into an animal, and bumping against the machine or container was highest in fall. The comparison of the container types showed that the risk of escape behavior, wing flapping, and bumping into an animal was increased when loading with the SmartStack container, but the risk of bumping against the machine or container was reduced. For animals of the husbandry system "Outdoor Climate," the risk of bumping into an animal and bumping against the machine or container was reduced. In addition, we found an influence of the examined parameters on the loading-related injuries. Increased escape behavior reduced the risk of severe injuries (fractures, dislocations, epiphysiolyses). Wing flapping and bumping against the machine or container increased the risk of hematomas and abrasions. The risk of hematomas was increased if broilers bumped into conspecifics. In summary, our analysis revealed that the behavior and impacts on the animals during loading are influenced by each of the analyzed factors and that they in turn can affect loading-related injuries.

Behavior of Piglets in an Observation Arena before and after Surgical Castration with Local Anesthesia.

Surgical castration of piglets is generally recognized as a painful procedure, but there is currently no gold standard for the assessment of pain behavior in piglets. However, pain assessment is essential for evaluating the effectiveness of local anesthetics. In this study, we investigated the efficacy of four local anesthetics in terms of pain relief during and after surgical castration in three sequential study parts. To do so, we filmed 178 piglets before the applied procedures, after injection of the local anesthetic, and up to 24 h after castration (five observation times in total) in an observation arena and compared their behavior before and after castration and between treatments and control groups. The results showed significant differences in the behavior of the piglets before and after castration and between the sham-castrated control group and the control group castrated without anesthesia. The different local anesthesia treatment groups showed diverging differences to the control groups. The most frequently shown pain-associated behaviors of the piglets were changes in tail position and hunched back posture. We observed a reduction but no complete elimination of the expressed pain-associated behaviors after local anesthesia. Several behavioral changes-such as changes in tail position, hunched back posture or tail wagging-persisted until the day after castration. Owing to the limited duration of the effects of the local anesthetics, local anesthesia did not influence long-term pain.

Urine risk factors in children with calcium kidney stones and their siblings.

Calcium nephrolithiasis in children is increasing in prevalence and tends to be recurrent. Although children have a lower incidence of nephrolithiasis than adults, its etiology in children is less well understood; hence, treatments targeted for adults may not be optimal in children. To better understand metabolic abnormalities in stone-forming children, we compared chemical measurements and the crystallization properties of 24-h urine collections from 129 stone formers matched to 105 non-stone-forming siblings and 183 normal, healthy children with no family history of stones, all aged 6 to 17 years. The principal risk factor for calcium stone formation was hypercalciuria. Stone formers have strikingly higher calcium excretion along with high supersaturation for calcium oxalate and calcium phosphate, and a reduced distance between the upper limit of metastability and supersaturation for calcium phosphate, indicating increased risk of calcium phosphate crystallization. Other differences in urine chemistry that exist between adult stone formers and normal individuals such as hyperoxaluria, hypocitraturia, abnormal urine pH, and low urine volume were not found in these children. Hence, hypercalciuria and a reduction in the gap between calcium phosphate upper limit of metastability and supersaturation are crucial determinants of stone risk. This highlights the importance of managing hypercalciuria in children with calcium stones.

Zoonotic agents in small ruminants kept on city farms in southern Germany.

Sheep and goats are popular examples of livestock kept on city farms. In these settings, close contacts between humans and animals frequently occur. Although it is widely accepted that small ruminants can carry numerous zoonotic agents, it is unknown which of these agents actually occur in sheep and goats on city farms in Germany. We sampled feces and nasal liquid of 48 animals (28 goats, 20 sheep) distributed in 7 city farms and on one activity playground in southern Germany. We found that 100% of the sampled sheep and 89.3% of the goats carried Shiga toxin-producing Escherichia coli (STEC). The presence of Staphylococcus spp. in 75% of both sheep and goats could be demonstrated. Campylobacter spp. were detected in 25% and 14.3% of the sheep and goats, respectively. Neither Salmonella spp. nor Coxiella burnetii was found. On the basis of these data, we propose a reasonable hygiene scheme to prevent transmission of zoonotic agents during city farm visits.

Which animal-to-feeding-place ratio at time-controlled hay racks is animal appropriate? Preliminary analysis of stress responses of horses.

Introduction: Automated feeding systems offer numerous advantages for animals and humans, but the associated benefits and risks can often only be seen under practical conditions. The space allowance (~80 cm per horse) at time-controlled hay racks for horses in group housing, unlike at partitioned feeding stands or transponder-controlled feed stations, currently falls below the required individual distance between the horses, which can result in a high level of aggression between the horses due to the lack of partitions between them. Hence, a feed-through at a time-controlled hay rack cannot be equated with a feeding place. In this preliminary study, we therefore aimed to determine the minimum animal-to-feeding-place ratio (AFR) at time-controlled hay racks that would provide adequate individual distances between the horses. Methods: To do so, we assessed behavioral and physiological parameters (via video behavioral observations and salivary cortisol measurements) of up to 28 horses in a loose housing system. Over 2 observation days per treatment, four AFRs were investigated in a balanced sequence: 1:1.2, 1:2, 1:3, and C (single feeding in familiar surroundings as a control). Results: We found that the horses expressed less aggressive behavior, especially those behaviors with a high risk of injury such as biting and kicking, when there were three times as many openings as there were horses at time-controlled hay racks, as compared with only 20% more openings or twice as many openings as there were horses [lineal mixed model: F (3, 4) = 7.411; adjusted R 2 = 0.733; p (AFR_1:2) = 0.06, p (AFR_1:3) = 0.02, p (AFR_C) = 0.01]. The salivary cortisol levels during feeding decreased more strongly with more generous AFRs [p (AFR_metric) = 0.02]. The factors hierarchy and individual showed no influence. In contrast, the day of the experiment and the associated weather conditions, despite randomized selection, influenced both the behavioral and the physiological parameters. Discussion: The results of this preliminary study indicate that the investigated time-controlled hay racks must provide at least three times as many feeding places as there are horses to ensure that neighboring horses can keep their individual distance and stress-free feeding is possible. Further studies on more farms and different types as well as arrangement of hay racks are proposed.

Effects of Stocking Rate and Environmental Enrichment on the Ontogeny of Pecking Behavior of Laying Hen Pullets Confined in Aviary Compartments during the First 4 Weeks of Life.

The objective of this study was to describe the ontogeny of the severe feather pecking (SFP), gentle feather pecking (GFP), aggressive pecking (AP), and enrichment pecking (EP) of non-beak-trimmed Lohmann Brown (LB)-pullets during the first 4 weeks of life (observation on 1st, 8th, 15th, 22nd, and 29th days of life) while they were kept within the compartments of a commercial rearing aviary (without access to a litter area). All chicks were placed into compartments of the middle tier of the aviary on the 1st day of life. On the 10th day of life, half of the chicks of each compartment were moved into the compartments of the lower tier. The aviary floor was covered with chick paper before the placement of the chicks and fully or partially removed from the 15th day of life onwards. The pecking behavior patterns were observed in three experimental groups (EG): NE (not enriched): group size until/after 10th day of life: 230/115; 120.8/60.4 birds/m2, no enrichment; EL (enriched, low stocking rate): group size until/after 10th day of life: 203/101, 106.6/53.6 birds/m2; and EH (enriched, high stocking rate): group size until/after 10th day of life: 230/115;120.8/60.4 birds/m2, both pecking stones and blocks as enrichment) in two rearing periods. For each pecking behavior pattern, an independent regression model with the parameters EG, chick paper, observation day, and functional area was estimated. GFP, SFP, and EP increased with age during the observation. The AP rate was highest in all EGs on the first day of life and decreased during the observation period. A pairwise comparison of NE (high stocking rate without enrichment) with EH (high stocking rate with enrichment and with EL (low stocking rate with enrichment) showed a significant effect of the EG on pecking behavior, with more SFP, AP, and GFP in NE. There were no differences between EL and EH, indicating that the provision of pecking materials had more influence than the stocking rate. However, we presumed that the difference between the stocking rates were too small to observe an effect. AP, SFP, and GFP were significantly higher on wired slats, as compared to the perches and the vicinity of the enrichment materials. The enrichment materials were suitable and intensely used by the pullets. The provision of pecking blocks and pecking stones was recommended as a preventive measure from the first day of life onwards for pullets housed in commercial rearing aviaries. There was no effect of reduced stocking rate, most likely due to the low variation in stocking rates.

Evaluation of small mammal pet supplies offered in German retail under animal welfare aspects.

German retailers offer a large variety of accessories for pets. However, not all products are suitable for pet husbandry. Several articles can negatively influence the wellbeing of pets or cause injuries, but empirical studies that evaluate accessories for small pets under animal welfare aspects are rare. In the present study, we assessed articles manufactured or sold in Germany in the product categories pet cages, hay racks, running wheels, exercise balls, harnesses and leashes, tube systems, and hamster bedding. To do so, we searched 28 German websites, visited 50 pet shops and 13 home improvement and garden centers on site and afterwards examined the animal welfare compliance of the products according to various evaluation criteria. Most of the examined products were rated not suitable for pet husbandry and were animal-welfare-adverse. This result applies to 86.1% (n = 87) of the 101 assessed running wheel models, 82.7% (n = 172) of the 208 assessed pet cage models and 55.6% (n = 40) of the 72 assessed hay rack models. The articles in the product categories exercise balls, harnesses and leashes, tube systems, and hamster bedding were also found unsuitable due to animal welfare concerns. Furthermore, we found clear shortcomings regarding article declarations. In some cases, relevant product information (e.g., dimensions) were missing, or the presented information was too general (e.g., rodent cage). Improperly declared pet accessories make it difficult for pet owners to decide whether a product is suitable or unsuitable for the species they keep. A declaration duty for manufacturers of pet products could ensure that German retailers only offer properly declared pet accessories and facilitate the decision for pet owners to purchase products appropriate for the pets they keep. Furthermore, a voluntary product certification for manufacturers would allow retailers to check the animal welfare compliance of articles before including them in their assortment. If a product is unsuitable for pet husbandry because it does not meet the set requirements, it must be considered animal-welfare-adverse and removed from the assortment. As done for the Austrian "animal welfare label," an independent, qualified third party could do the certification.

A comparison of two manual catching methods of broiler considering injuries and behavior.

The aim of this study was to compare the 2 manual catching methods in terms of injuries and behavior. Throughout 12 loadings on practical farms with the same standard, 1 container each was caught using the one-legged (1LCM) and 1 using the two-legged catching method (2LCM). The animals were filmed during loading to evaluate their behavior and subsequently examined regarding injuries. Wing flapping was observed more frequently in broilers caught with the 1LCM than 2 LCM. Carrying animals with neighbors (1 neighbor: P < 0.001; 2 neighbors: P < 0.001) and a grasping position at or above the tarsal joint (P < 0.001; P < 0.054) reduced wing flapping in both methods. A short grasping duration (P = 0.004), settling the broilers into the crate (P = 0.005) and avoiding striking the broilers against the crate (P < 0.001) reduced the occurrence of wing flapping. About 1.1% of 1LCM and 0.43% of 2LCM broilers were diagnosed with an epiphysiolysis. Catching with the 1LCM (P = 0.042), loading in lower crates (low vs. middle: P = 0.005; low vs. high: P = 0.008), a longer catching duration (p = 0.025) and female broilers (P = 0.007) had a higher chance for epiphysiolysis. Broilers loaded in lower crates (P = 0.007) and ones which showed more wing flapping (P = 0.015) had a higher chance for hematomas. A higher loading duration led to a higher risk of hematomas (prevalence: 1.5%) and a prevalence of 1.0% of broilers with severe injury in 2LCM in a simultaneously performed study (mechanical loading vs. 2LCM), in which manual loadings of entire barns were evaluated. This may be caused by fatigue of the workers. In summary, the catching method or number of grasped legs is not the decisive factor, but the compliance and implementation of the identified risks and careful handling of the animals are determining factors to reduce wing injuries caused by loading and wing flapping.

Validation of an automatic scoring system for the assessment of hock burn in broiler.

This study aimed to develop and validate a camera vision score that could detect macroscopic alterations of the hock, to identify errors and to assess possible factors that could influence the assessment. Two hundred hocks in the first (calibration) phase and 500 hocks in the second (validation) phase were collected at slaughter, visually assessed, placed back into the evisceration line and assessed by a camera system with 2 software systems. The size of the alteration in percent (%) measured by the camera system was evaluated ("camera score", CS). Additionally, temperature, humidity, and light intensities were measured. In the calibration phase, threshold values of camera scores for respective macro scores were defined and performance measures evaluated. In the validation phase, the generated threshold values were validated, occurring errors, as well as possible impacts of climatic factors analyzed. The results showed that the generated thresholds predict the camera score values at which the respective macro score has the highest probability of appearance. Small hock burn lesions ≤0.5 cm have the highest probability at a camera score of ≥0.2 (original CS) or ≥0.1 (updated CS), and lesions >0.5 cm have the highest probability at a camera score of ≥0.7 (original CS) or ≥1.1 (updated CS). Large lesions (>0.5 cm) are more reliably identified by the system than small lesions. The risks of errors in assessing reference areas and lesions showed a correct identification of lesions to be the most probable result even if the reference area is not correctly identified. The probability of a correct identification of lesions by the camera system was slightly higher (not significant) with the updated software (risk = 0.66 [0.62-0.70]) than with the original software (risk = 0.63 [0.58-0.67]). Automatic assessment systems at slaughter could be adjusted to the presented threshold values to classify hock burn lesions. Software adaptations can improve the performance measures of diagnosis and reduce the probability of errors.

Individual plumage and integument scoring of laying hens on commercial farms: correlation with severe feather pecking and prognosis by visual scoring on flock level.

Various plumage and integument scoring methods are commonly used to deduce the occurrence of severe feather pecking and cannibalism in laying hens. The aim of our study was to provide evidence of correlations between the occurrence of severe feather pecking and our individual plumage scoring system used under practical conditions on commercial farms with non-beak-trimmed and beak-trimmed layers (study I). In second step, we aimed to verify whether the results of the elaborate individual scoring may be predicted with a visual scoring method based on the total body scores of groups of birds (study II). For study I we observed the pecking behavior and performed an individual plumage scoring at the beginning, in the middle, and at the end of a laying period on 8 commercial farms. For study II we performed both an individual and a visual plumage scoring on 49 flocks on 45 farms at the beginning of the laying period and on 43 flocks on 41 farms at the end of the laying period. Spearman's Rho revealed a correlation of the mean feather pecking rate with the total plumage score, the neck-back plumage score, and the total cannibalism score in all observation periods. A high feather pecking rate was correlated with severe plumage damage and the frequent occurrence of skin injuries. We conclude that both the total plumage score and the neck-back plumage score constitute a reliable indicator of the occurrence of severe feather pecking in the flocks assessed in this study. The results of study II suggest that the percental assessment of plumage damage on flock level in 3 categories ("visual score") leads to a good prognosis of the actual, individually assessed plumage score. Therefore, the application (and documentation) of the visual score on a regular basis can provide a good evaluation of the development of the plumage condition of the flock. The visual score presented in this study is suggested as a suitable instrument for self-evaluation programs on farms.

Evaluation of Two Injection Techniques in Combination with the Local Anesthetics Lidocaine and Mepivacaine for Piglets Undergoing Surgical Castration.

The present study evaluated the effects of two injection techniques in combination with lidocaine or mepivacaine for piglets undergoing castration. To improve ease of use, a cannula with side holes (one-step fenestrated (F)) was invented to simultaneously deliver a local anesthetic into the testis and scrotum and was compared to a two-step injection technique. The distribution of a lidocaine/contrast agent mixture using the two methods was examined using computed tomography. Piglets were randomly divided into treatment groups: handling, castration without pain relief and castration after lidocaine or mepivacaine injection using the one-step F or two-step method. Acute physiological responses to noxious stimuli were evaluated by measuring the mean arterial blood pressure (MAP), heart rate (HR) and nocifensive movements. Fos protein expression in the spinal dorsal cord was semi-quantitatively analyzed. Both injection techniques achieved similar distribution patterns. The one-step F method was faster and easier. Injection was not associated with significant changes in MAP or HR, but Mepi1 and NaCl elicited significantly increased nocifensive movements. Both techniques significantly reduced MAP and nocifensive movements when the spermatic cords were cut, regardless of the local anesthetic type. Compared to NaCl, only the lidocaine treatments significantly reduced HR during skin incision. Lido2 significantly reduced Fos protein expression.

Nociception-Induced Changes in Electroencephalographic Activity and FOS Protein Expression in Piglets Undergoing Castration under Isoflurane Anaesthesia.

The objective of this study was to investigate the electroencephalographic reaction pattern and FOS protein expression in male piglets undergoing surgical castration under light isoflurane anaesthesia with or without local anaesthesia. The experiment was conducted under isoflurane anaesthesia to exclude the effect of the affective components of pain on the measurements. Changes in the oscillatory activity of the cerebral cortex over a 90 s period after noxious stimulation or simulated interventions were analysed. FOS expression was determined postmortem by performing immunohistochemistry in the dorsal horn of the spinal cord. The analysis of the response to an interdigital pinch revealed a biphasic reaction pattern in the electroencephalogram (EEG) that similarly was observed for the surgical stimuli during the castration procedure in the group without analgesia. This EEG response was attenuated or altered by the application of local anaesthetics. Immunohistochemical staining for FOS indicated a lower expression in the handling and in three local anaesthetic groups than in the animals castrated without pain relief. The findings indicate that EEG and FOS expression may serve as indicators for nociception in piglets under light isoflurane anaesthesia. A lower activation of nociceptive pathways occurs during castration after the application of local anaesthetics. However, EEG and FOS analyses should be combined with additional parameters to assess nociception, e.g., haemodynamic monitoring.

Qualitative and quantitative analyses of polysomnographic measurements in foals.

Veterinary and human medicine are still seeking a conclusive explanation of the function of sleep, including the change in sleep behaviour over the course of an individual's lifetime. In human medicine, sleep disorders and abnormalities in the electroencephalogram are used for prognostic statements, therapeutic means and diagnoses. To facilitate such use in foal medicine, we monitored 10 foals polysomnographically for 48 h. Via 10 attached cup electrodes, brain waves were recorded by electroencephalography, eye movements by electrooculography and muscle activity by electromyography. Wireless polysomnographs allowed us to measure the foals in their home stables. In addition, each foal was simultaneously monitored with infrared video cameras. By combining the recorded data, we determined the time budgeting of the foals over 48 h, whereby the states of vigilance were divided into wakefulness, light sleep, slow-wave sleep and rapid-eye-movement sleep, and the body positions into standing, suckling, sternal recumbency and lateral recumbency. The results of the qualitative analyses showed that the brain waves of the foals differ in their morphology from those previously reported for adult horses. The quantitative data analyses revealed that foals suckle throughout all periods of the day, including night-time. The results of our combined measurements allow optimizing the daily schedule of the foals according to their sleep and activity times. We recommend that stall rest should begin no later than 9.00 p.m. and daily stable work should be done in the late afternoon.

Saliva and Blood Cortisol Measurement in Bottlenose Dolphins (Tursiops truncatus): Methodology, Application, and Limitations.

A central task of zoos and aquaria is the frequent and accurate assessment of their animals' welfare. Recently, important steps have been made, such as the introduction of animal welfare evaluation tools and welfare decision trees. To determine animal welfare, it is not only important to collect life history data, such as longevity and reproductive success, but also for experienced observers or caretakers to conduct behavioral observations on a regular basis to assess animals' emotional state. To physiologically validate welfare observations, glucocorticoid levels are usually assessed, as they are a common indicator of stress. While, for many animals, these levels can be easily determined via fecal or hair samples, for cetaceans, the levels are usually determined via blood samples. As blood samples cannot be taken very frequently and the process may cause stress to the animals (if the samples are not taken following medical training), other techniques, such as the measurement of health biomarkers (especially cortisol, which can be measured in saliva), have become the focus of cetacean stress research. However, there are two problems associated with saliva measurements in cetaceans: saliva might either be diluted with pool water or be contaminated by fodder fish, as frozen fish usually contains high levels of cortisol. In our study, we investigated how saliva cortisol levels are connected to blood cortisol levels and how saliva cortisol can be influenced by fodder fish. We examined saliva and blood samples in eleven bottlenose dolphins (Tursiops truncatus) kept in an outdoor and indoor facility in Germany. Furthermore, we assessed the cortisol levels of different kinds of fodder fish. Our data show that, although saliva cortisol values are elevated under stress and arousal, they seem not to be correlated with blood cortisol values. We also show that, after feeding, saliva cortisol values are increased up to 100-fold. Our results suggest that saliva cortisol measurements in dolphins have to be conducted and considered with care, as they can easily be contaminated. Moreover, it is important to use the right laboratory method in order to specifically detect cortisol; in our study, we conducted reliable tests, using LC-MS/MS.

Feather Pecking in Non-Beak-Trimmed and Beak-Trimmed Laying Hens on Commercial Farms with Aviaries.

Severe feather pecking (SFP) is a major animal welfare problem in layers. It results in pain and injuries in the affected animal. It was the aim of this study to gain insight into the actual pecking behavior of laying hens kept on commercial farms with flock sizes common in practice. We observed aggressive pecking and SFP in non-beak-trimmed and beak-trimmed flocks of laying hens and investigated possible influencing factors. The study took place on eight conventional farms in Germany with aviaries, including three farms with a free range and a winter garden, one with a free range and one with a winter garden. Pecking behavior was observed during three observational periods (OPs): OP 1, at the peak of the laying period between the 28th and 33rd week of life; OP 2, in the middle of the laying period between the 42nd and 48th week of life; and OP 3, at the end of the laying period between the 63rd and 68th week of life in one laying period. Videos were analyzed using behavior sampling and continuous recording. We found that SFP occurred in all flocks, but the pecking rate differed significantly between the flocks. SFP correlated positively with the number of hens per square meter of usable area, with statistical significance in the litter area (r = 0.564; p = 0.045). The multivariate analysis revealed that access to a winter garden or free range significantly reduced the SFP rate on perches (p = 0.001). The stocking density (number of birds per usable square meter) had a significant influence on the SPF rate in the nest-box area (p = 0.001). The hybrid line had a significant effect on the SFP rate on perches and in the nest-box area (p = 0.001 each). Lohmann Brown hens in mixed flocks had a higher SFP rate (significant in OP 2) than those in homogeneous flocks, indicating that mixed flocks may be a risk factor for SFP. Lohmann Brown hens pecked significantly less than Dekalb White hens in the litter area (p = 0.010) and in the nest-box area (p = 0.025) and less than Lohmann Selected Leghorn hens in the litter area (p = 0.010). Lohmann Brown and Lohmann Selected Leghorn hens showed increasing SFP rates during the laying period. All hybrid lines had significantly higher SFP rates in the litter area, followed by the nest-box area and perches. These findings emphasize the importance of providing enough litter, litter areas and environmental enrichment. We found a significant positive correlation between aggressive pecking and SFP-in OP 1: rho (Spearman) = 0.580, p < 0.001; OP 2: rho = 0.486, p = 0.002; and OP 3: rho = 0.482, p = 0.002 (n = 39) -indicating that SFP may lead to a higher stress level in the flock. Beak trimming reduced pecking rates but did not entirely prevent SFP. Instead of subjecting chicks to this potentially painful procedure, reasons for SFP should be addressed. In conclusion, our data suggest a positive influence of a lower stocking density and the provision of a winter garden or free range for additional space. The hybrid line had a significant influence on the feather-pecking rate on perches and the nest-box area. Aggressive pecking and severe feather pecking correlated positively. We assume that vigorous and painful AP were an additional stress factor, especially in non-beak-trimmed flocks, leading to more SFP in due course. Beak trimming had a reducing effect on SFP. However, our results showed that non-beak-trimmed flocks could be kept without major outbreaks of SFP.

Welfare and Performance of Three Turkey Breeds-Comparison between Infrared Beak Treatment and Natural Beak Abrasion by Pecking on a Screed Grinding Wheel.

Feather pecking and cannibalism are behavioral disorders that cause animal-welfare-relevant and economic problems. To mitigate these problems, the beaks of conventionally reared turkeys are usually already trimmed in the hatcheries. To find an alternative to beak trimming, we conducted this study with male turkeys of three breeds: B.U.T. 6, B.U.T. Premium and, Auburn (200 turkeys per breed). Half of the birds had infrared-trimmed beaks; the other half had intact beaks. For each treatment combination (breed, beak status), 25 turkeys were housed in one section. A screed grinding wheel was installed in each feed pan of the non-beak-trimmed turkeys as of week six to facilitate natural beak abrasion until slaughter. Eight randomly selected turkeys per section were regularly examined to record injuries, plumage condition, and beak dimensions. In addition, 96 beaks from randomly slaughtered birds were examined macroscopically and histologically. The results concerning injuries and plumage condition showed in most cases no differences between the beak-trimmed turkeys and the ones provided with the blunting disks. The histological examinations revealed alterations in only the beak-trimmed birds. We can conclude that the blunting method smoothens the beak during feeding and thus may be a possible alternative to beak trimming.