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Feline infectious peritonitis (FIP) is a fatal disease in cats caused by infection with feline coronavirus (FCoV). Despite severe inflammatory changes, defense mechanisms fail to achieve virus clearance. Some studies focused on various immune evasion mechanisms, but none of these studies elucidated the inefficacy of the complement system, which is one major player in FIP-associated immune pathogenesis. This study aimed to investigate the involvement of complement-regulating factors (CRFs). CRFs help modulate the immune response and prevent host tissue damage. Archived tissue samples from 31 deceased, FIP-affected cats were evaluated using multiplex immunohistochemistry for the spatial expression of the complement-regulating factors CD46 and CD59 in association with FIP lesions and their colocalization with complement-activating factor C1q and membrane attack complex C9 in relation to the presence and proximity of FCoV-infected cells. The FIP lesions of all 31 cats exhibited marked expression of both complement-regulating factors in proximity to FCoV-infected macrophages. Moreover, their expression in all 31 animals was significantly lower than the expression of the complement-activating factors C1q and C9 compared to areas farther distal to FCoV-infected cells. In conclusion, FCoV-infected macrophages in cats with FIP appear to use autocrine and paracrine expression of complement-regulating factors in their immediate environment to shield themselves from destruction by the complement system.

A 24/7 AI sound-based coughing monitoring system was applied in combination with oral fluids (OFs) and bioaerosol (AS)-based screening for respiratory pathogens in a conventional pig nursery. The objective was to assess the additional value of the AI to identify disease patterns in association with molecular diagnostics to gain information on the etiology of respiratory distress in a multimicrobially infected pig population. Respiratory distress was measured 24/7 by the AI and compared to human observations. Screening for swine influenza A virus (swIAV), porcine reproductive and respiratory disease virus (PRRSV), Mycoplasma (M.) hyopneumoniae, Actinobacillus (A.) pleuropneumoniae, and porcine circovirus 2 (PCV2) was conducted using qPCR. Except for M. hyopneumoniae, all of the investigated pathogens were detected within the study period. High swIAV-RNA loads in OFs and AS were significantly associated with a decrease in respiratory health, expressed by a respiratory health score calculated by the AI The odds of detecting PRRSV or A. pleuropneumoniae were significantly higher for OFs compared to AS. qPCR examinations of OFs revealed significantly lower Ct-values for swIAV and A. pleuropneumoniae compared to AS. In addition to acting as an early warning system, AI gained respiratory health data combined with laboratory diagnostics, can indicate the etiology of respiratory distress.

This study aimed to assess the efficacy of a single intra-articular injection of autologous stromal vascular fraction (SVF) in dogs with chronic lameness due to advanced elbow osteoarthritis (OA) that were unresponsive to conventional drug therapy. In this clinical, prospective, non-blinded, single-center study, twenty-three dogs received autologous SVF derived from falciform adipose tissue. Primary outcome measures over the six-month study period included clinical-orthopedic and radiographic examinations, objective gait analysis and validated owner questionnaires. In 19 of 23 joints, no progression of OA was visible radiographically. Peak vertical force improved significantly at three months and vertical impulse at six months after the injection compared to baseline. Over 33% of dogs demonstrated treatment-related improvements in lameness based on objective gait analysis. Owner questionnaires indicated significant improvement in clinical signs throughout the study period and 26% of dogs showed treatment-related improvements in pain scores according to the Canine Brief Pain Inventory. No side effects were reported. These findings suggest that autologous regenerative cell therapy may provide a promising treatment option for dogs with advanced OA that do not respond to conventional drug therapy. However, the treatment did not improve the clinical symptoms in all dogs, so it cannot be recommended for all patients.

In recent years several studies established the diagnostic value of anti-Müllerian hormone (AMH) in companion animals. However, less is known about physiological AMH concentrations in young individuals highlighting the necessity to apply the diagnostic findings to this group. The aim of this study was to determine the AMH values of healthy male and female dogs between the age of 8 and 48 weeks, tomcats under 8 weeks and up to 48 weeks of age and queens between 2 to 12 weeks of age. In total, 96 blood samples were collected. Anti-Müllerian hormone was measured in all samples and testosterone was measured in the oldest age group of the males in both species. The hormones were analyzed using a human based chemiluminescence immune assay. Overall, AMH concentrations were higher in males than in females (p < 0.001). According to the AMH concentration there was no difference in males, but queens had significant higher AMH concentrations than bitches (p < 0.001). AMH remained high in males up to week 24 and decreased significantly thereafter (tomcats: p = 0.015; male dogs: p = 0.013), which correlated with an increase in testosterone levels for male dogs only. In bitches, AMH remained below the detection limit until the week 16 and slightly increased subsequently. In queens, AMH was detectable from the beginning with a significant increase in the older age group (p = 0.003). Half of the cats in the older age group even approached the chemiluminescence immune assay’s upper limit. The results show that female cats secrete AMH much earlier than female dogs in which AMH secretion begins just shortly before the start of the puberty. In the male animals, the decrease in AMH concentration around puberty was similar in dogs and cats, but a correlation with the increase of testosterone was only observed in dogs. Further research is required to determine the origin of the high AMH concentrations in female kittens and the lack of correlation between testosterone and AMH concentrations in male kittens.

BackgroundSevere and complex angular limb deformities in dogs require accurate morphological assessment using diagnostic imaging to achieve successful orthopedic surgery. Computed tomography (CT) is commonly used to overcome projection errors in two-dimensional angular measurements of dog hindlimb alignment. Three-dimensional volume rendering (VR) techniques permit virtual positioning and variable projection, but the final CT-image that defines the projection plane for angular measurements remains two-dimensional.ObjectiveWe wanted to develop a true three-dimensional open-source technique to measure the alignments of the hind limbs of dogs in CT scanners.MethodsWe developed an open-source 3D Slicer plug-in, to perform angular measurements using vector calculations in three-dimensional space. In 113 CT-scans of canine pelvic limbs, femoral torsion, femoral varus, femorotibial rotation, tibial torsion, tibial varus and tibiotalar rotation angles were calculated and compared to an already validated technique using VoXim®.ResultsReference points were identified and measurements were possible in the 113 acquisitions. The greatest difference between the two techniques was 1.4° at only one tibial torsion angle. Mean values for all Bland-Altman plots did not show significant differences and were less than 0.07° for all comparisons.DiscussionBased on these results we considered angular measurements of canine hind limb alignment in CT scans using the 3D Slicer extension program sufficiently accurate for clinical orthopedic and surgical purposes in veterinary medicine.ConclusionWith our open-source 3D Slicer extension software, we provide a free accessible tool for veterinary orthopedic surgeons and thus we hope to improve angular measurements in CT-scans of canine hind limb deformities through true three-dimensionality.

Background: Cataracts resulting from equine recurrent uveitis (ERU) or other forms of uveitis are usually associated with rapid progression. ERU is the most common ocular disease cause of blindness and cause of cataracts in horses. The necessity for the posterior capsulorhexis (PC) during phacoemulsification (PE) is controversial. This study aimed to evaluate vision and complications after PE combined with pars plana vitrectomy (PPV) in horses with uveitis-associated cataracts and compare the PE technique with and without posterior capsulorhexis. Methods: Thirty-two eyes of 28 horses with uveitis-associated cataracts aged 14 months to 19.6 years were treated with PE-PPV under identical conditions. Twenty-three eyes of 21 horses were affected by an ERU-associated (ERU group), and nine eyes of 7 horses were affected by cataracts related to uveitis with pathogenesis different to ERU (non-ERU group). PE-PPV was performed in 12 eyes of 10 horses (PC group) and 20 eyes of 18 horses without posterior capsulorhexis (NPC group). Follow-up examination was performed at a mean of 1.7 ± 1.8 years postoperatively (range: 1 month–6.4 years). Results: In the period up to 1 month postoperatively, 17/20 (85%) NPC-eyes and 8/12 (67%) PC-eyes (total: 25/32 [78%]) were visual. From 1–6 months postoperatively, 16/20 (80%) NPC-eyes and 7/12 (58.3%) PC-eyes (total: 23/32 [72%]), and from 6–12 months, 7/11 (63.6%) NPC-eyes and 3/8 (37.5%) PC-eyes (total: 10/19 [52.6%]) were visual. From 12–18 months postoperatively, 3/7 (42.9%) NPC-eyes and 2/9 (22.2%) PC-eyes (total: 5/16 [31.3%]), and from 18–24 months, 3/8 (37.5%) NPC-eyes and 1/8 (12.5%) PC-eyes (total: 4/16 [25%]) were visual. After 24 months postoperatively, 2/7 (28.6%) NPC-eyes and 1/8 (12.5%) PC-eyes (total: 3/15 [20%]) were visual. Despite the higher number of visual eyes in the NPC group at each time point, differences were not significant. No obvious differences regarding postsurgical vision were observed between the ERU- and non-ERU groups at each time point. In the overall population, a significant decrease in the number of eyes with postoperative active uveitis was observed during the follow-up examinations (p < 0.001). A significant increase in the number of eyes that were blind due to retinal detachment was observed in the overall patient population as the examination period progressed (p < 0.001). Retinal detachment was the sole long-term cause of blindness. Conclusions: In horses diagnosed with uveitis-associated cataracts and treated with PE-PPV, no persistent active uveitis was observed in the present study during follow-up examinations. However, the proportion of eyes that were blind due to retinal detachment increased. Whilst PE-PPV may prevent postsurgical persistent active uveitis and remove lens opacity, the prognosis for a visual outcome is guarded. A superior outcome in postsurgical vision was observed in the NPC group. However, caution is required when interpreting these results due to several factors that affect the independent comparison of the surgical groups.

IntroductionDogs with medial patellar luxation can be affected by pelvic limb deformities whose corrective osteotomies and associated biomechanical rebalancing might provide higher success rates than standard surgical procedures limited to the stifle joint. In bilaterally affected canine patients, comparison with the contralateral normal limb is impossible. Reference values are useful for orthopedic decision-making. Inconsistency of published reference values might depend on methodology or canine breed. We hypothesized that canine pelvic limb alignment is breed-specific.MethodsCT scans of 42 pelvic limbs of dog breeds predisposed for medial patellar luxation, with an orthotopic patellar position and stability were studied. Several angleswere measured with an open-source 3D Slicer plugin using vector calculations. The breeds were compared with a general linear model with a Bonferonni adjustment using SPSS.ResultsChihuahuas, Pomeranians, Jack Russel Terriers, Pugs, French Bulldogs, Maltese were examined. In the order of the listed breeds, the angles were as follows: 28.3°±10.7°, 20.1°±2.9°, 35.4°±6.9°, 32.8°±3.0°, 19.0°±7.1°, 26.6°±5.3° for the antetorsion, 5.3°±1.8°, 2.8°±2.8°, 8°±4.4°, 3.8 °±3.1°, 4.7°±3.3°, 2.3°±3.3° for the femoral varus, of −5.5°±6.2°, 1.1°±4.1°, −5.2°±9.5°, 6.1°±8.0°, −0.1°±5.9°, −9.2°±4.7° for the tibial torsion, 2.0°±2.9°, 2.1°±2.7°, 6.4°±6.8°, 0.0°±5.7°, 3.0°±5.8°, 8.8°±8.6° for the tibial valgus, 1.2°±10.4°, 1.8°±3.4°, −1.7°±4.9°, −1.7°±9.4°, 5.1°±8.8°, −0.2°±8.6° for the femorotibial rotation and −3.4°±2.2°, 1.1°±4.1°, −2.8°±3.4°, −5.2°±4.0°, −2.1°±4.4°, −5.4°±3.7° for the tibiotalar rotation. There were significant differences between breeds in femoral torsion, femoral varus, and tibial torsion angles, but no significant differences in tibial valgus, femorotibial, and tibiotalar rotation angles.DiscussionOur hypothesis is therefore partially correct. Our results are limited to small dogs prone to medial patellar luxation and might not be generalized. To establish robust reference values larger case numbers and more breeds should be evaluated. In conclusion, canine pelvic limb alignment reference values for small dogs with a predisposition for medial patellar luxation should be considered breed-specific.

IntroductionCanine tibial alignment is determined by two-dimensional angular measurements, and tibial torsion is challenging. Aim of the study was the development and evaluation of a CT technique to measure canine tibial varus and torsion angles independent from positioning and truly three-dimensional.Materials and methodsA bone-centered 3D cartesian coordinate system was introduced into the CT-scans of canine tibiae and aligned with the anatomical planes of the bone based on osseous reference points. Tibial torsion, and varus (or valgus) angles were calculated based on geometric definition of projection planes with VoXim® medical imaging software using 3D coordinates of the reference points. To test accuracy of the tibial torsion angle measurements, CT scans of a tibial torsion model were performed in 12 different hinge rotation setups ranging from the normal anatomical situation up to +/ 90° and compared to goniometer measurements. Independency of tibial positioning on the CT scanner table was evaluated in 20 normal canine tibiae that were scanned in a position parallel to the z-axis and two additional off-angle double oblique positions having 15° and 45° deviation in direction of the x- and y-axes. Angular measurements in oblique positions were compared with the normal parallel position by subtraction. Precision was tested using clinical CT scans of 34 canine patients with a clinical diagnosis of patellar luxation.ResultsAccuracy testing in the tibial torsional deformity model revealed a difference of 0.2° demonstrated by Passing-Bablok analysis and Bland–Altman-Plots. Testing for independency from tibial positioning resulted in mean differences less than 1.3°. Precision testing in clinical patients resulted in coefficients of variation for repeated measurements of 2.35% (intraobserver agreement) and 0.60% (interobserver agreement) for the tibial torsion angle, and 2.70% (intraobserver agreement) and 0.97% (interobserver agreement) for the tibial varus (or valgus) angle.DiscussionThe technique is lacking determination of bone deformities in the sagittal plane, and demonstration of accuracy in severe complex bone deformities in multiple planes.In conclusion, we developed a method to measure canine tibial torsional and varus or valgus deformities, that calculates in 3D space, and we demonstrated its accuracy in a torsional deformity model, and its precision in CT data of clinical patients.

IntroductionIn small animal orthopedics, angular measurements in the canine femur are often applied in clinical patients with bone deformities and especially in complex and severe cases. Computed tomography (CT) has been shown to be more precise and accurate than two-dimensional radiography, and several methods are described. Measurement techniques evaluated in normal bones must prove accuracy in deformed bones in clinical settings.ObjectivesThe goals of our study were to evaluate the accuracy of canine femoral torsion angle measurements in a femoral torsional deformity model and to test repeatability and reproducibility of canine femoral neck inclination, torsion, and varus angle measurements in CT datasets of dogs applying a CT-based technique using a three-dimensional (3D) bone-centered coordinate system.Materials and methodsFor precision testing, femoral torsion, femoral neck inclination, and femoral varus angles were measured in CT data of 68 canine hind limbs by two operators, and their results were compared. For accuracy testing, a femoral torsional deformity model was preset from 0° to +/−90° with a goniometer and scanned. Torsion angles were measured in the CT data and compared to the preset value.ResultsIn the femoral torsion model, the Bland–Altman plots demonstrated a mean difference of 2.11°, and the Passing–Bablok analysis demonstrated a correlation between goniometer and CT-based measurements. In the clinical CT scans, intra- and interobserver agreement resulted in coefficients of variation for repeated measurements (%) between 1.99 and 8.26 for the femoral torsion, between 0.59 and 4.47 for the femoral neck inclination, and between 1.06 and 5.15 for the femoral varus angles.DiscussionEvaluation of femoral malformations with torsional deformities is the target area of this technique. Further studies are required to assess its value in different types, degrees, and combinations of osseous deformities and to establish normal reference values and guidelines for corrective osteotomies.ConclusionBased on the results of this study, the accuracy of the torsion angle measurements and the precision of inclination, torsion, and the varus angle measurements were considered acceptable for clinical application.

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.

IntroductionMeasurement of torsional deformities and varus alignment in the canine femur is clinically and surgically important but difficult. Computed tomography (CT) generates true three-dimensional (3D) information and is used to overcome the limitations of radiography. The 3D CT images can be rotated freely, but the final view for angle measurements remains a subjective variable decision, especially in severe and complex angular and torsional deformities. The aim of this study was the development of a technique to measure femoral angles in a truly three-dimensional way, independent of femoral positioning.MethodsTo be able to set reference points in any image and at arbitrary positions of the CT series, the 3D coordinates of the reference points were used for mathematical calculation of the angle measurements using the 3D medical imaging Software VoXim®. Anatomical reference points were described in multiplanar reconstructions and volume rendering CT. A 3D bone-centered coordinate system was introduced and aligned with the anatomical planes of the femur. For torsion angle measurements, the transverse projection plane was mathematically defined by orthogonality to the longitudinal diaphyseal axis. For varus angle measurements, the dorsal plane was defined by a femoral retrocondylar axis. Independence positioning was tested by comparison of angle measurement results in repeated scans of 13 femur bones in different parallel and two double oblique (15/45°) positions in the gantry. Femoralvarus (or valgus), neck version (torsion), and inclination angles were measured, each in two variations.ResultsResulting mean differences ranged between –0.9° and 1.3° for all six determined types of angles and in a difference of

ABSTRACT: 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.

Abstract Horizontal transmission of Mycoplasma suis via parenteral exposure during standard practices or through bites during fightings have been identified as key epidemiological routes. However, as knowledge gaps on other potential shedding and transmission routes exist, the present study combines both laboratory experiments and field surveys to gain new insights into the epidemiology of porcine haemotrophic mycoplasmas. Splenectomised pigs were orally inoculated with a M. suis field strain and investigated for clinical signs related to infectious anaemia of pigs (IAP) and the presence of M. suis in blood, urine and saliva samples by qPCR. All blood samples were negative for M. suis and animals did not show obvious clinical signs of IAP throughout the entire study period. Additionally, urine, nasal and saliva samples from sows of conventional piglet producing farms and semen samples from a boar stud revealed no detection of M. suis and ‘Candidatus Mycoplasma haemosuis’ by qPCR. Thus, the results indicate that blood-independent transmission routes might be of minor relevance under field conditions.

Abstract In the three decades since its inception, the sex-sorting technology has progressed significantly. However, field studies report conflicting findings regarding reproductive outcomes. Therefore, we conducted this meta-analysis of all trials published between 1999 and 2021. Non-return rates after 24 or 60 d (NRR 24/60), pregnancy, calving, abortion, and stillbirth rates were compared after AI with sex-sorted vs non-sorted sperm. Additionally, the impact of recent developments in the sex-sorting technology was assessed. Of 860 studies found, 45 studies (72 trials) provided extractable data and were included. Overall, the results of this meta-analysis provided evidence that the NRR 24/60 was diminished by 13%, pregnancy rates were reduced by 23% (25% cows, 21% heifers) and calving rates were reduced by 24% when using sex-sorted sperm. Enhancing the dosage to 4 million sex-sorted sperm/straw (including recent improvements, high vs low dose) as well as using fresh sex-sorted sperm (sorted vs non-sorted) increased pregnancy rate ratios by 7 percentage points. The refinement of the sex-sorting technology after 2015 resulted in a lowered reduction of pregnancy and calving rate of 19% and 23%, respectively. Whereas abortion rates were similar, the stillbirth of male calves was increased by 6.3%.

Abstract 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.

Epilepsy is a common neurological disorder affecting 0.6–0.75% of dogs in veterinary practice. Treatment is frequently complicated by the occurrence of drug-resistant epilepsy and cluster seizures in dogs with idiopathic epilepsy. Only few studies are available to guide treatment choices beyond licensed veterinary drugs. The aim of the study was to compare antiseizure efficacy and tolerability of two add-on treatment strategies in dogs with drug-resistant idiopathic epilepsy. The study design was a prospective, open-label, non-blinded, comparative treatment trial. Treatment success was defined as a 3-fold extension of the longest baseline interseizure interval and to a minimum of 3 months. To avoid prolonged adherence to a presumably ineffective treatment strategy, dog owners could leave the study after the third day with generalized seizures if the interseizure interval failed to show a relevant increase. Twenty-six dogs (mean age 5.5 years, mean seizure frequency 4/month) with drug-resistant idiopathic epilepsy and a history of cluster seizures were included. Dogs received either add-on treatment with pregabalin (PGB) 4 mg/kg twice daily (14 dogs) or a dose increase in levetiracetam (LEV) add-on treatment (12 dogs). Thirteen dogs in the PGB group had drug levels within the therapeutic range for humans. Two dogs in the PGB group (14.3%; 2/14) and one dog in the LEV group (8.3%; 1/12) achieved treatment success with long seizure-free intervals from 122 to 219 days but then relapsed to their early seizure frequency 10 months after the study inclusion. The overall low success rates with both treatment strategies likely reflect a real-life situation in canine drug-resistant idiopathic epilepsy in everyday veterinary practice. These results delineate the need for research on better pharmacologic and non-pharmacologic treatment strategies in dogs with drug-resistant epilepsy.

Abstract Background Long-term cyclic tensile testing with equine palmar/plantar tendons have not yet been performed due to problems in fixing equine tendons securely and loading them cyclically. It is well established that the biomechanical response of tendons varies during cyclic loading over time. The aim of this study was to develop a clamping device that enables repetitive cyclic tensile testing of equine superficial digital flexor tendon for at least 60 loading cycles and for 5 min. Results A novel cryo-clamp was developed and built. Healthy and collagenase-treated pony SDFTs were mounted in the custom-made cryo-clamp for the proximal tendon end and a special clamping device for the short pastern bone (os coronale). Simultaneously with tensile testing, we used a biplanar high-speed fluoroscopy system (FluoKin) to track tendon movement. The FluoKin system was additionally validated in precision measurements. During the cyclic tensile tests of the SDFTs, the average maximal force measured was 325 N and 953 N for a length variation of 2 and 4 % respectively. The resulting stress averaged 16 MPa and 48 MPa respectively, while the modulus of elasticity was 828 MPa and 1212 MPa respectively. Length variation of the metacarpal region was, on average, 4.87 % higher after incubation with collagenase. The precision of the FluoKin tracking was 0.0377 mm, defined as the standard deviation of pairwise intermarker distances embedded in rigid bodies. The systems accuracy was 0.0287 mm, which is the difference between the machined and mean measured distance. Conclusion In this study, a good performing clamping technique for equine tendons under repetitive cyclic loading conditions is described. The presented cryo-clamps were tested up to 50 min duration and up to the machine maximal capacity of 10 kN. With the possibility of repetitive loading a stabilization of the time-force-curve and changes of hysteresis and creep became obvious after a dozen cycles, which underlines the necessity of repetitive cyclical testing. Furthermore, biplanar high-speed fluoroscopy seems an appropriate and highly precise measurement tool for analysis of tendon behaviour under repetitive load in equine SDFTs.

The stillbirth, mummification, embryonic death, and infertility (SMEDI) syndrome is most commonly associated with porcine parvovirus 1 (PPV1) infections. Little is known about the occurrence of coinfections with SMEDI-associated pathogens and the associations among these pathogens. In our study, we included 40 SMEDI-affected litters from 18 different farms. In total, 158 out of 358 available fetuses from diagnostic transmittals were selected by systematic random sampling and examined for PCV2, PCV3, PPV1, and Leptospira spp. by q-PCR. Results from diagnostic materials showed the following results: in eleven farms, PCV2 was present; in nine farms, PPV1 was present; in five farms, PCV3 was present; and in two farms, Leptospira spp. was present. The detection of Leptospira spp. was significantly associated with a PCV2 coinfection (OR: 26.3; p < 0.001). PCV3 positivity resulted in a reduced probability of detecting PCV2 in the corresponding fetus (OR: 0.078; p = 0.008). Fetal maceration was associated with Leptospira spp. detection (OR: 8.6; p = 0.003), whereas mummification (p = 0.047), reduced crown-rump length (p < 0.001), and bodyweight (p = 0.001) of fetuses were significantly associated with PPV1 and PCV2 coinfection and thus, presumably, a shorter time to death after infection, indicating an enhanced negative effect on the development of fetuses with PCV2 + PPV1 coinfection.

Animal welfare assurance is of great importance as dairy farming is recently under increasing pressure to meet societal and commercial expectations. Therefore, this study aims to determine the current welfare situation of dairy cattle housed in tie-stalls and free stalls in South Tyrol (Eastern Italian Alps) in order to evaluate the need for establishment of an animal welfare assurance programme. For reasons of research economy, a protocol was used for data collection that would also be applicable for practical use in an animal welfare assurance scheme. Analyses of resource-based and animal-based indicators recorded in 204 farms in North and South Tyrol (1891 dairy cows) reveal some important animal welfare problems mainly related to the provision of resources and the prevalence of skin alterations especially in tie-stall barns, which are still widely spread in mountain areas. Hence, the implementation of an animal welfare assurance scheme is urgently needed to reflect public concerns through regular and standardised monitoring of welfare indicators and continuous encouragement of improvements in dairy cattle welfare towards predefined targets. Concerning tie-stalls, interventions in stall design as well as the selective use of local breeds best adapted to the mountainous conditions appear to be appropriate measures to optimise dairy cattle health and welfare. These findings substantiate the high value of the data that would be collected as part of the assurance programme to gain insights, which could be used in preventive and corrective health plans to improve welfare-friendliness in dairy farming of South Tyrol.Highlights An adverse effect on animal welfare in tie-stalls was pointed out, even though this housing system is still widely spread in the Alpine region. An animal welfare assurance programme for dairy cattle is urgently needed in South Tyrol, where some welfare problems currently exist. Regular and standardised monitoring of indicators encourages improvements in dairy cattle health and welfare.

The implementation of an animal welfare assurance programme for dairy cattle in South Tyrol (Eastern Italian Alps) faces particular feasibility constraints due to the outstanding volume of travel associated with routine on-farm audits of remote mountain farms. Therefore, this study aims to estimate the inter-rater reliability of the expert’s and farmers’ welfare outcome assessment regarding recommendations to involve milk producers in animal welfare assurance within South Tyrolean dairy farming. A formal training programme containing a classroom session and an on-farm observation became mandatory for all 188 participating farmers, which was offered by the expert, applied as reference standard. On-farm data collected on the farmers’ cows (data set of 1719 dairy cows) were compared at animal level. Cohen’s kappa, respectively, weighted kappa, examined for several welfare indicators, range from slight to moderate agreement (κ = 0.018 − 0.416; κω = 0.163 − 0.310). These findings are further confirmed by results at farm level (ICC = 0.018 − 0.577). Continuous repeatability checks as part of routine audits are therefore proposed to substantially reduce the variability between the raters and to avoid significant bias in the welfare outcome assessment. In this way, the competence for regular and standardised monitoring could be increasingly transferred to dairy farmers in order to reduce the need for costly and time-consuming inspections by external auditors, which are in long-term perspective also harmful to the alpine environment. Additionally, the promotion of welfare assessment as an instructive management tool would intensify farmers’ commitment to the assessment process.Highlights Farmers’ self-assessment of welfare outcomes is cost-effective and eco-friendly, but reliability must be ensured. Inter-rater reliability of welfare outcome assessment by an expert and farmers presented a slight to moderate level. Repeatability assessment at regular intervals is proposed to reduce data variability and, thus, prevent bias in the welfare outcome assessment.

Abstract Background Dogs with hypoadrenocorticism (HA) frequently show signs of gastrointestinal disease (SGD). The prevalence of dogs presented for chronic SGD with HA is unknown. Objectives The aims of this study were to determine the prevalence of HA in dogs with chronic SGD and to identify clinical and laboratory variables for HA in this population. Animals One hundred fifty‐one dogs with chronic SGD. Methods In this multicentered prevalence study a standardized workup was performed in prospectively enrolled dogs with SGD > 3 weeks duration. Basal serum cortisol concentration was measured in every dog with ACTH stimulation test (ACTHST) if basal serum cortisol concentration was

Abstract Background Leptospirosis is a widespread zoonosis and has been recognized as a re-emerging infectious disease in humans and dogs, but prevalence of Leptospira shedding in dogs in Thailand is unknown. The aim of this study was to determine urinary shedding of Leptospira in dogs in Thailand, to evaluate antibody prevalence by microscopic agglutination test (MAT) and enzyme-linked immunosorbent assay (ELISA), and to assess risk factors for Leptospira infection. In Northern, Northeastern, and Central Thailand, 273 stray (n = 119) or client-owned (n = 154) dogs from rural (n = 139) or urban (n = 134) areas were randomly included. Dogs that had received antibiotics within 4 weeks prior to sampling were excluded. No dog had received vaccination against Leptospira. Urine was evaluated by real-time polymerase chain reaction (PCR) specific for lipL32 gene of pathogenic Leptospira. Additionally, urine was cultured for 6 months in Ellinghausen-McCullough-Johnson-Harris (EMJH) medium. Antibodies were measured by ELISA and MAT against 24 serovars belonging to 15 serogroups and 1 undesignated serogroup. Risk factor analysis was performed with backwards stepwise selection based on Wald. Results Twelve of 273 (4.4%; 95% confidence interval (CI): 2.0–6.8%) urine samples were PCR-positive. In 1/273 dogs (0.4%; 95% CI: 0.01–1.1%) Leptospira could be cultured from urine. MAT detected antibodies in 33/273 dogs (12.1%; 95% CI: 8.2–16.0%) against 19 different serovars (Anhoa, Australis, Ballum, Bataviae, Bratislava, Broomi, Canicola, Copenhageni, Coxi, Grippotyphosa, Haemolytica, Icterohaemorrhagiae, Khorat, Paidjan, Patoc, Pyrogenes, Rachmati, Saxkoebing, Sejroe). In 111/252 dogs (44.0%; 95% CI: 37.9–50.2%) immunoglobulin M (IgM) and/or immunoglobulin G (IgG) antibodies were found by ELISA. Female dogs had a significantly higher risk for Leptospira infection (p = 0.023). Conclusions Leptospira shedding occurs in randomly sampled dogs in Thailand, with infection rates comparable to those of Europe and the USA. Therefore, the potential zoonotic risk should not be underestimated and use of Leptospira vaccines are recommended.

Infections with feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) occur worldwide and are among the most important infectious diseases in cats. The aim of the present study was to determine the prevalence of FIV and FeLV infection in healthy outdoor cats in North, Northeast and Central Thailand. So far, a study on retrovirus prevalence of healthy cats in Thailand in a larger geographic area has not been published yet. In addition, risk factors for FIV and FeLV infections were evaluated. Two hundred sixty healthy cats were prospectively recruited. They originated from 13 locations in North, Northeast, and Central Thailand and were presented for either preventive health care and/or neutering. In each cat, a physical examination was performed to confirm health status. FIV and FeLV status was determined using a commercial rapid enzyme-linked immunosorbent assay (ELISA) (SNAP Combo Plus FeLV/FIV, IDEXX). Risk factors were analyzed by binary logistic regression analysis. Samples of 15/260 (5.8%) cats were positive for FIV antibodies, and 11/260 (4.2%) samples were positive for FeLV antigen. One of the 260 (0.4%) cats was positive for both, FIV and FeLV infection. In binary logistic regression analysis, no parameter was associated with a higher risk for FeLV infection. However, cats had a significantly (p = 0.025) higher risk for FIV infection when they were 2 years or older. FIV and FeLV infections occur in healthy cats in North, Northeast and Central Thailand, but prevalence was lower than expected. No risk factors for FeLV infection were detected, but risk for FIV infection increases with age.

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.

Large-animal models for Duchenne muscular dystrophy (DMD) are crucial for the evaluation of diagnostic procedures and treatment strategies. Pigs cloned from male cells lacking DMD exon 52 (DMDΔ52) exhibit molecular, clinical and pathological hallmarks of DMD, but die before sexual maturity and cannot be propagated by breeding. Therefore, we generated female DMD+/− carriers. A single founder animal had 11 litters with 29 DMDY/−, 34 DMD+/− as well as 36 male and 29 female wild-type offspring. Breeding with F1 and F2 DMD+/− carriers resulted in an additional 114 DMDY/− piglets. With intensive neonatal management, the majority survived for 3-4 months, providing statistically relevant cohorts for experimental studies. Pathological investigations and proteome studies of skeletal muscles and myocardium confirmed the resemblance to human disease mechanisms. Importantly, DMDY/− pigs displayed progressive myocardial fibrosis and increased expression of connexin-43, associated with significantly reduced left ventricular ejection fraction, at 3 months. Furthermore, behavioral tests provided evidence for impaired cognitive ability. Our breeding cohort of DMDΔ52 pigs and standardized tissue repositories provide important resources for studying DMD disease mechanisms and for testing novel treatment strategies.

Abstract Introduction Equine asthma represents a naturally occurring animal model for human allergic neutrophilic asthma. Inhalative nanoparticle‐bound cytosine‐phosphate‐guanosine (CpG‐GNP) immunotherapy, independent of specific allergens, has already shown promising clinical and immunological results in previous studies and offers the possibility to treat the underlying cause of the disease. This study analyses the relationship between dose and response, and evaluates a possible long‐term effect. Methods In the prospective, randomised, double‐blind clinical field study, 29 horses suffering from equine asthma received 10 inhalation treatments with either 187.5 µg CpG‐GNP (CpG single dose [CpGsd]; n = 11), 375 µg CpG‐GNP double dose (CpG double dose [CpGdd]; n = 9) (q48h for 20 days) or 1600 µg beclomethasone (n = 9) (q24h for 10 days). Each horse was examined three times: before the treatment (I), immediately after the 10 inhalations (II), and 8 weeks after the final inhalation (III). The three groups were compared according to clinical and laboratory parameters. The study examined the sustainability of the long‐term effect of the treatment after 8 weeks, as well as the tolerability of the formula as a double dose. Results The CpGsd resulted in a significant improvement in 82% of the parameters, the CpGdd in 72%. In the long‐term evaluation, the CpGsd showed a significant improvement in 100% of the parameters in comparison to the initial values, the CpGdd in 67%. On the immunological level, the bronchoalveolar lavage revealed a significant reduction of IL‐4, IL‐8, and interferon‐γ. Conclusion Both CpG groups displayed significant improvements in clinical and laboratory parameters, especially regarding the long‐term effect of CpGsd. Doubling the CpG dose did not result in any improvement in comparison to the original single dose. On the immunological level, an anti‐inflammatory, as well as an immunomodulatory effect, apart from a Th2‐dominated immune response, could be observed. This immunomodulatory inhalation treatment could indicate a new possibility for human allergic asthma therapy.

Abstract Background Despite appropriate medical therapy, many horses with equine recurrent uveitis continue to suffer from recurrent bouts of inflammation. Surgical intervention via the pars plana vitrectomy or suprachoroidal cyclosporine implant placement may control and/or prevent recurrences, however, these procedures may be contraindicated, unavailable, or declined by an owner. Thus, an effective adjunctive treatment option may help to improve the clinical outcomes in those situations. There are several anecdotal reports on the use of intravitreal gentamicin injections, but to date, no data evaluating the complication rate and/or treatment effect following this treatment have been published. Thus, the aim of this prospective study was to describe the intravitreal gentamicin injection technique, describe the associated peri-injection (within 24 h) and post-injection (30 to 780 days) complications, and to report the effects of the injection on the clinical signs of uveitis. Additionally, evaluation of the systemic and ocular Leptospira-status, and its effect on the treatment outcome was performed. A total of 86 horses of various ages, breeds, and gender presenting with recurrent or persistent uveitis were treated via intravitreal injection of 4 mg of undiluted gentamicin (0.04 ml, Genta 100, 100 mg/ml in 35 horses) or preservative-free gentamicin (0.05 ml, 80 mg/ml in 52 horses) under sedation and local anesthesia. All 86 horses were observed for immediate peri-injection and post-injection complications. Response to therapy was evaluated in 59 of the 86 horses (follow-up: 30 to 780 days). Results Peri-injection complications consisted of subconjunctival (26/86; 30.2%) or intracameral hemorrhage (4/86; 4.7%); both of which completely resolved within 5 days. Post-injection complications consisted of cataract formation/maturation (5/59 horses, 8.5%) and diffuse retinal degeneration (3/59 eyes 5.1%). The majority of horses 52/59 (88.1%) with a minimum follow-up period of 30 days were controlled (absence of recurrent or persistent inflammation) at their last recheck examination. Recurrent inflammation was documented in 5/59 (8.5%) horses and persistent inflammation was diagnosed in 2/59 (3.4%) horses. Conclusions Intravitreal injection of low-dose gentamicin shows promise at controlling different types and stages of uveitis. The ability of intravitreal injections of low-dose gentamicin (4 mg) to control persistent and recurrent inflammation warrants further investigation.

Severe feather pecking (SFP) is a behavioral disorder, for which there are multifactorial reasons. Various aspects of pullet and laying-hen husbandry—including housing conditions, management, feeding, and genetics—must be considered, to prevent negative outcomes, such as severe plumage damage, skin injuries, and high mortality rates due to SFP. The aim of this study was to identify housing and management factors in the occurrence of feather pecking, so as to reduce the risk of this behavioral disorder in non-beak-trimmed laying hens on commercial farms with aviaries. Beak trimming may reduce the extent of plumage damage, but it does not prevent feather pecking and cannibalism. Thirty non-beak-trimmed flocks (16 in the first, and 14 in the second laying period) were investigated on 16 commercial farms in Germany. Each flock was visited twice during rearing, and three times during the laying period. During each visit, individual plumage and integument scoring were performed, and data were collected, regarding the housing and management conditions of the flocks. To analyze the influence of management and housing on the plumage condition of the rearing and laying flocks, models were calculated, using univariate multifactorial analysis. In the rearing period, high stocking density and poor litter quality were significant risk factors in plumage damage due to SFP. In the laying period, a lack of free range, poor litter quality, insufficient enrichment, and plumage damage during rearing were significant risk factors for the development of SFP. An individual risk analysis of pullet and layer farms is therefore strongly recommended, to prevent outbreaks of SFP and cannibalism, especially in non-beak-trimmed birds.

BackgroundThe impact of dietary phosphorus (P) excess, especially on renal and cardiovascular health, has been investigated in several species, but little is known in dogs.ObjectiveThe aim of this study was to examine effects of different P sources on concentration and postprandial kinetics of selected parameters of P homeostasis in dogs.MethodsEight beagles received one control diet (P 0.5% dry matter [DM]) and three high P diets (poultry meal, NaH2PO4, and KH2PO4; P 1.7% DM) for 18d. Urine samples were collected pre- and postprandially while faeces were collected quantitatively for 5d and analysed for minerals. On day 18, blood was sampled 1h pre- and 0.5, 1, 1.5, 2, 3, 5 and 7h postprandially.ResultsPi (KH2PO4, NaH2PO4) but not organic P caused an increased apparent P digestibility and significantly influenced kinetics of serum FGF23, parathyroid hormone, P, CrossLaps and bonespecific alkaline phosphatase, demonstrating a disrupted calcium (Ca) and P homeostasis with potential harm for renal, cardiovascular and skeletal health.ConclusionsResults of feeding Pi to dogs indicate distinct disturbances of Ca and P metabolism, in contrast to organic sources. The use of Pi in food can therefore not be considered as safe. Further research, especially on dose and long-term effects, is warranted.

Vertical transmission is a consistently discussed pathway of porcine circovirus type 2 (PCV2) and porcine circovirus type 3 (PCV3) transmission in pigs. To evaluate the presence of PCV2 and PCV3 in piglets, we collected tissue samples from 185 piglets that were crushed within the first week of life from 16 farms located in Germany and Austria. Pooled samples consisting of thymus, inguinal lymph node, myocardium, lung and spleen were examined for PCV2 and PCV3 by qPCR. Furthermore, oral fluid samples (OFS) from grow–finish pigs were collected and examined the same way. In piglets, PCV2 was highly prevalent (litters: 69.4%; piglets: 61.6%), whereas PCV3 prevalence was low (litters: 13.4%; piglets: 13.0%). In total, 72.6% and 67.2% of all collected OFS were PCV2 or PCV3 positive, respectively. Sow vaccination against PCV2 was identified as a protective factor concerning PCV2 in piglets (OR: 0.279; CI: 0.134–0.578; p < 0.001), whereas the porcine reproductive and respiratory syndrome virus (PRRSV) vaccination of sows was identified as a protective factor concerning PCV3 in piglets (OR: 0.252 CI: 0.104–0.610; p = 0.002). Our results show that PCV2, but not PCV3, is ubiquitous in suckling piglets and that early PCV3 infections might be modulated by PRRSV–PCV3 interaction. However, the ubiquitous nature of both viruses in older pigs could be confirmed.

Introduction of new imaging modalities for the equine brain have refocused attention on the horse as a natural model for ethological, neuroanatomical, and neuroscientific investigations. As opposed to imaging studies, strategies for equine neurodissection still lack a structured approach, standardization and reproducibility. In contrast to other species, where adapted protocols for sampling have been published, no comparable guideline is currently available for equids. Hence, we developed a species-specific slice protocol for whole brain vs. hemispheric dissection and tested its applicability and practicability in the field, as well as its neuroanatomical accuracy and reproducibility. Dissection steps are concisely described and depicted by schematic illustrations, photographs and instructional videos. Care was taken to show the brain in relation to the raters' hands, cutting devices and bench surface. Guidance is based on a minimum of external anatomical landmarks followed by geometric instructions that led to procurement of 14 targeted slabs. The protocol was performed on 55 formalin-fixed brains by three groups of investigators with different neuroanatomical skills. Validation of brain dissection outcomes addressed the aptitude of slabs for neuroanatomical studies as opposed to simplified routine diagnostic purposes. Across all raters, as much as 95.2% of slabs were appropriate for neuroanatomical studies, and 100% of slabs qualified for a routine diagnostic setting. Neither autolysis nor subfixation significantly affected neuroanatomical accuracy score, while a significant negative effect was observed with brain extraction artifacts. Procedure times ranged from 14 to 66 min and reached a mean duration of 23.25 ± 7.93 min in the last of five trials in inexperienced raters vs. 16 ± 2.83 min in experts, while acceleration of the dissection did not negatively impact neuroanatomical accuracy. This protocol, derived analogously to the consensus report of the International Veterinary Epilepsy Task Force in dogs and cats, allows for systematic, quick and easy dissection of the equine brain, even for inexperienced investigators. Obtained slabs feature virtually all functional subcompartments at suitable planes for both diagnostic and neuroscientific investigations and complement the data obtained from imaging studies. The instructive protocol and brain dissection videos are available in Supplementary Material.

Abstract Introduction New therapeutic strategies to modulate the immune response of human and equine allergic asthma are still under extensive investigation. Immunomodulating agents stimulating T‐regulatory cells offer new treatment options beyond conventional symptomatic treatment or specific immunotherapy for human and equine allergic airway diseases, with the goal of a homoeostatic T‐helper cell balance. The aim of this study was to evaluate the effects of a nebulized gelatin nanoparticle‐CpG formulation (CpG‐GNP) with and without specific allergens for the treatment of spontaneous allergic equine asthma as a model for human asthma. Methods Twenty equine asthma‐affected horses were treated either with CpG‐GNP alone or CpG‐GNP with allergens. Two specific allergens were selected for each horse based on history and an in‐vitro test. Each horse received seven administrations of the respective nebulized composition and was examined before treatment, immediately after and 6 weeks after the treatment course. Results Clinical parameters such as breathing rate, indirect interpleural measurement, arterial blood gases, amount of tracheal mucus and percentage of neutrophils and cytokines in tracheal washes and serum samples were evaluated. Treatment with CpG‐GNP alone as well as in combinations with relevant allergens resulted in clinical improvement of nasal discharge, breathing rate, amount of secretion and viscosity, neutrophil percentage and partial oxygen pressure directly after and 6 weeks after treatment. There were no significant differences between the two treatments in clinical parameters or local cytokine profiles in the tracheal wash fluid (IL‐10, IFN‐g, and IL‐17). IL‐4 concentrations decreased significantly in both groups. Conclusion Nonspecific CpG‐GNP‐based immunotherapy shows potential as a treatment for equine and possibly also human allergic asthma.

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.

Contamination of fresh produce with human pathogens poses an important risk for consumers, especially after raw consumption. Moreover, if microorganisms are internalized, no removal by means of further hygienic measures would be possible. Human pathogenic bacteria identified in these food items are mostly of human or animal origin and an adaptation to this new niche and particularly for internalization would be presumed. This study compares a plant-internalized and an animal-borne Salmonella enterica subsp. enterica serovar Choleraesuis aiming at the identification of adaptation of the plant-internalized strain to its original environment. For this purpose, a phenotypical characterization by means of growth curves under conditions resembling the indigenous environment from the plant-internalized strain and further analyses using Pulsed-field gel electrophoresis and Matrix-assisted laser desorption ionization time of flight spectrometry were assessed. Furthermore, comparative genomic analyses by means of single nucleotide polymorphisms and identification of present/absent genes were performed. Although some phenotypical and genetic differences could be found, no signs of a specific adaptation for colonization and internalization in plants could be clearly identified. This could suggest that any Salmonella strain could directly settle in this niche without any evolutionary process being necessary. Further comparative analysis including internalized strains would be necessary to assess this question. However, these kinds of strains are not easily available.

During a search for immunomodulatory receptors in the chicken genome, we identified a previously cloned chicken sequence as CRTAM homologue by its overall identity and several conserved sequence features. For further characterization, we generated a CRTAM specific mab. No staining was detectable in freshly isolated cell preparations from thymus, bursa, caecal tonsils, spleen, blood and intestine. Activation of splenocytes with recombinant IL-2 increased rapid CRTAM expression within a 2 h period on about 30% of the cells. These CRTAM(+) cells were identified as CD8(+) γδ T lymphocytes. In contrast, CRTAM expression could not be stimulated on PBL with IL-2, even within a 48 h stimulation period. As a second means of activation, T cell receptor (TCR) crosslinking using an anti-αβ-TCR induced CRTAM on both PBL and splenocytes. While CRTAM expression was again rapidly upregulated on splenocytes within 2 h, it took 48 h to reach maximum levels of CRTAM expression in PBL. Strikingly, albeit the stimulation of splenocytes was performed with anti-αβ-TCR, CRTAM expression after 2 h was mainly restricted to CD8(+) γδ T lymphocytes, however, the longer anti-TCR stimulation of peripheral blood lymphocytes (PBL) resulted in CRTAM expression on αβ T lymphocytes. In order to characterize the potential ligand we cloned and expressed chicken Necl-2, a member of the nectin and nectin-like family which is highly homologous to its mammalian counterpart. Three independent assays including a reporter assay, staining with a CRTAM-Ig fusion protein and a cell conjugate assay confirmed the interaction of CRTAM with Necl-2 which could also be blocked by a soluble CRTAM-Ig fusion protein or a CRTAM specific mab. These results suggest that chicken CRTAM represents an early activation antigen on CD8(+) T cells which binds to Necl-2 and is upregulated with distinct kinetics on αβ versus γδ T lymphocytes.