Altered vocalization rate during the estrous cycle in dairy cattle.

Correct detection of estrus is a problem in dairy herds. In practice, several procedures exist for detection of estrus besides conventional visual observation by humans. These procedures deliver very different results regarding detection of estrus. It is known that the calls of female mammals can contain information about reproductive status. It is also suspected that the vocalizations of cattle contain information about age, sex, dominance status, and stage in the estrous cycle. In the present study, a methodology for the continuous automatic recording of vocalization of heifers during the periestrous period is presented. It was shown in 10 tethered heifers that the estrous climax results in an increase in vocalization rate. Vocalization rate of heifers increased approximately 84% from d -2 to 0 (related to observed estrus) and approximately 59% from d -1 to d 0. After d 0, vocalization rate decreased about 79%. Increased vocalization was correlated with the visual observation of estrus by humans. We also found 2 different structures in the vocalization of heifers. The harmonic structure showed regular frequency bands, whereas the nonharmonic structure was noisy. The hypothesis that the disharmonic structure increases near the estrous climax was confirmed. Hence, it seems possible to get information about stage of the estrous cycle of dairy cattle by means of monitoring vocalization. The presented method of automatically detecting the rate of cattle vocalization (patent pending) could be used solely or in combination with other automated systems for detecting estrus and could considerably increase current estrus detection rates once its applicability can be demonstrated in nontethered cattle.

Effects of attention and rewarded activity on immune parameters and wound healing in pigs.

Challenging animals with a demanding situation they are able to cope with and where they are rewarded may be a source of positive emotions and possibly of increased welfare. In order to test if this results in changes of immunological parameters and wound healing, 56 pigs (7-20 weeks of age), housed in groups of 8 animals each, were successfully trained to recognize and localize an individual acoustic summons and to receive a small portion of feed as a reward. Immune reactions and the development of a standardized biopsy wound were compared to values of conventionally fed control groups of equal size and animal-to-feeding-place ratio (2:1). In the experimental animals a significantly higher concentration of IgG as well as an increased in vitro T-cell proliferation to ConA but a reduced LPS-induced proliferation of B-cells was found, while basal salivary cortisol concentrations were similar. Wound development was better in the experimental animals as measured by the area of the inflammatory corona. It decreased more rapidly in the experimental animals since the 5th day after biopsy and was significantly smaller than in the control groups. We conclude that environmental enrichment by equipment provoking attention and cognitive activity which is rewarded by feed may play a beneficial role for physical welfare of intensively housed pigs.

Colony stimulating factor (CSF) in human mixed lymphocyte cultures: effect on human and mouse progenitor cells.

Supernatants from human two-way MLC, poor and rich in monocytes, were tested for their ability to induce colony growth of human and mouse progenitor cells in semi solid agar. Colony stimulating factor (CSF), with activity in both systems, indicated that allogenic lymphocytes require monocytes to produce CSF. Whereas human marrow showed an early kinetics of production, the liberation of CSF active on mouse marrow cells exhibited a delayed kinetics. These results, combined with other evidence, suggest that human lymphocytes produce two different factors.

[Reaction of muscles to a stimulating environment--effects on the loin muscle (M. longissimus) of Landrace pigs]. / Reaktion der Muskulatur auf eine stimulierende Haltung--Befunde am Kotelettmuskel (M. longissimus) von Landrasse-Schweinen.

The effects of an enriched environment on growth, meat quality, and muscle biological traits were investigated using 64 castrated pigs (age range 7 to 20 weeks). Positive emotional appraisal was induced by a system that acoustically frequently called individuals out of a group (n=32) to a feeding station, where they were rewarded with small portions of feed. The results were compared with traditional fed pigs (Control; n = 32). There was no environmental effect on live weight and lean muscle percentage. However, the stimulating keeping regime reduced the intramuscular fat content of the M. longissimus by 0.2 % and increased the protein content by 0.5 %. Moreover, the drip loss of M. longissimus was 1.1 % lower in the treatment group. The analysis of muscle fibre traits (biopsy samples of M. longissimus) showed a 7 % higher proportion of oxidative muscle fibres in the pigs reared in the enriched environment. This fibre type showed a tendency to increased fibre areas. These findings indicate a higher oxidative capacity of the muscle. However, the activity of the inarker enzyme citrate synthase (CS) was not influenced and the activity of lactate dehydrogenase (LDH) was even increased compared to the control animals.

Origins of descending projections to the medulla oblongata and rostral medulla spinalis in the urodele Salamandra salamandra (amphibia).

Descending projections to the medulla oblongata and rostral medulla spinalis have been examined in the urodele Salamandra salamandra with retrograde horseradish peroxidase tracing. Ipsilateral projections originate from the striatum and the nucleus ventrolateralis thalami and reach the medulla oblongata. The ipsilateral nucleus praeopticus magnocellularis reaches the medulla spinalis. The rostral part of the nucleus tuberculi posterioris projects to the ipsilateral medulla oblongata; its caudal part projects further caudally. Tectal efferents and the efferents of the nucleus praetectalis profundus project bilaterally, the nucleus praetectalis superficialis, nucleus mesencephalicus nervi trigemini, torus semicircularis, nucleus Darkschewitsch, and nucleus fasciculi longitudinalis medialis project ipsilaterally to the medulla oblongata. The nucleus mesencephalicus nervi trigemini, nucleus fasciculi longitudinalis medialis, and tectal efferents reach the rostral medulla spinalis. The nucleus ruber projects mainly via the contralateral dorsolateral funiculus to the medulla spinalis. A largely crossed medullary projection arises in the nucleus dorsalis tegmenti pars anterior, a bilateral projection arises in the nucleus dorsalis tegmenti pars posterior, and an ipsilateral projection arises in the nucleus ventralis tegmenti pars anterior. Cerebellar and statoacoustic efferents descend to the medulla spinalis. The nucleus reticularis isthmi, superior, medius and inferior as well as the nucleus raphes exhibit spinal trajectories. The nucleus vestibularis magnocellularis projects bilaterally, the nucleus vestibularis medialis projects ipsilaterally spinalward. The supposed nucleus descendens nervi trigemini descends mainly contralaterally. A small spinal projection arises in the nucleus tractus solitarii. The results indicate that salamander brains display elaborate descending connections which are similar to those in other vertebrates despite their scarcely differentiated neuronal cytoarchitecture.

Localization of motoneurons innervating the extraocular muscles in Salamandra salamandra L. (Amphibia, Urodela).

The central innervation patterns of the extraocular muscles were investigated in the European fire salamander Salamandra salamandra L. by means of the horseradish peroxidase method. The ipsilateral portion of the nucleus nervi oculomotorii, which is located in the rostral ventral tegmentum mesencephali, supplies the musculi recti inferior and medialis and the musculus obliquus inferior without a clear somatotopic representation of the motoneurons. The musculus rectus superior is innervated mainly by a contralateral portion of this nucleus. A definite nucleus Edinger-Westphal could not be recognized. The nucleus nervi trochlearis, which rostrally joins the nucleus nervi oculomotorii with a gap of only about 40 micron between the nuclei, is situated completely contralateral to the musculus obliquus superior supplied by it. The nucleus nervi abducentis, innervating the musculus rectus lateralis, and the nucleus accessorius nervi abducentis, supplying the musculus retractor bulbi, are found in the ipsilateral medulla oblongata and exhibit a large rostrocaudal extension from the eighth cranial nerve to the first root of the vagus nerve. Dendrites of the nucleus nervi oculumotorii and of the nucleus accessorius nervi abducentis extend into neuropil areas receiving primary sensory afferents.

On the presence of nucleus ruber in the urodele Salamandra salamandra and the caecilian Ichthyophis kohtaoensis.

The presence of nucleus ruber in urodeles and caecilians (amphibia) was investigated. For that purpose, horseradish peroxidase was applied to the rostral spinal cord, the medulla oblongata at various levels and the dorsolateral funiculus. Whereas Salamandra salamandra possesses a rubrospinal tract, it is absent in the limbless caecilian Ichthyophis kohtaoensis.

Binocular afferents to the salamander pretectum mediate rotation sensitivity of cells selective for visual background motions.

The majority (85%) of background motion-sensitive pretectal cells in salamanders was found to be binocularly driven. In 70% of the units the binocularity establishes a rotation selectivity with respect to background pattern-movements. This results in a considerable augmentation of the discharge rate when pattern movement is perceived simultaneously in the temporonasal direction by the contralateral eye and nasotemporally by the ipsilateral eye. The response is depressed when the pattern movement is seen in the same direction by both eyes. It is concluded that the rotation-sensitive cells are mainly excited by contralateral retinal afferents selective for temporonasal movements and inhibited by direct or indirect ipsilateral afferents with the same type of direction selectivity.

Directional selectivities of visual afferents to the pretectal neuropil in the fire salamander.

Single-unit recordings from visual afferents in the pretectal neuropil of Salamandra salamandra were performed to characterize the properties of direction specific units which are probably involved in the optokinetic reflex. It was shown that afferents from the contralateral eye were more sharply tuned than those from the ipsilateral eye. However, the majority of both were selective for temporonasal movements.

Effects of weaning and restraint stress on glucocorticoid receptor binding capacity in limbic areas of domestic pigs.

Changes in glucocorticoid receptor (GR) binding in different brain areas were investigated in neonatal and adult pigs exposed to psychological stress (weaning) and a physical stressor (repeated snaring). The GR binding was significantly decreased 4 days after weaning in both the hippocampus and the amygdala, but there were no changes in the hypothalamus. Repeated snaring of adult pigs resulted in a significant diminished GR binding only in the hippocampus.

Quantitative distribution of descending tectal efferent cells in salamanders.

In salamanders of the species Salamandra salamandra, Pleurodeles waltl, and Plethodon jordani, tectal efferent neurons projecting to the bulbar reticular formation show a stereotyped distribution as revealed by the horseradish peroxidase (HRP) method. The density of these cells increases toward caudal tectal levels reaching its maximum at the isthmic region. Neurons with ipsilaterally descending axons constitute 60-87% and are concentrated in the dorsomedial and ventrolateral sector of the tectal hemisphere, with only few neurons in the intermediate part. The contralaterally descending neurons are concentrated in the lateral sector. This pattern is assumed to reflect a motor map in the coordinates of the salamander neck muscle system.

Topographic relationships of ipsi- and contralateral visual inputs to the rostral tectum opticum in the salamander Plethodon jordani indicate the presence of a horopter.

Topographies of ipsilateral and contralateral retinal inputs to the optic tectum were studied by means of electrophysiological multi-unit recordings from the superficial layers. It was found that the nasotemporal coordinate of the visual field is represented along a rostrocaudal axis on the contralateral tectal map and along a caudorostral axis on the ipsilateral map. Electrical stimulation of one tectal hemisphere and recording of the response in the other hemisphere revealed that the ipsilateral map is most probably established by an intertectal pathway. In one tectal hemisphere, inputs from the ipsilateral eye match those from the contralateral eye if stimuli are located at a certain distance at arbitrary horizontal angles within the binocular field. This distance varied from 11.8 to 29.5 mm between the 5 individuals examined.

Visual discrimination learning in dwarf goats and associated changes in heart rate and heart rate variability.

We studied visual discrimination learning in a group of Nigerian dwarf goats using a computer-based learning device which was integrated in the animals' home pen. We conducted three consecutive learning tasks (T1, T2 and T3), each of which lasted for 13 days. In each task, a different set of four visual stimuli was presented on a computer screen in a four-choice design. Predefined sequences of stimulus combinations were presented in a pseudorandom order. Animals were rewarded with drinking water when they chose the positive stimulus by pressing a button next to it. Noninvasive measurements of goats' heartbeat intervals were carried out on the first and the last 2 days of each learning task. We analysed heart rate (HR) and heart rate variability (HRV) of resting animals to study sustained physiological effects related to general learning challenge rather than acute excitement during an actual learning session. The number of trials to reach the learning criterion was 1000 in T1, when visual stimuli were presented to the goats for the first time, but decreased to 210 in T2 and 240 in T3, respectively. A stable plateau of correct choices between 70% and 80% was reached on Day 10 in T1, on Day 8 in T2 and on Day 6 in T3. We found a significant influence of the task and of the interaction between task and day on learning success. Whereas HR increased throughout T1, this relationship was inverted in T2 and T3, indicating different effects on the HR depending on how familiar goats were with the learning task. We found a significant influence of the task and the interaction between task and time within the task on HRV parameters, indicating changes of vagal activity at the heart. The results suggest that changes in HR related to learning were predominantly caused by a withdrawal of vagal activity at the heart. With regard to nonlinear processes in heartbeat regulation, increased deterministic shares of HRV indicated that the animals did not really relax until the end of T3. Comparing changes of HR and HRV in T3 and in a subsequent postexperiment (PE), we assume a positive effect of such cognitive challenges once the task had been learned by the animals.

An automated training device for pattern discrimination learning of group-housed gerbils.

The setup, designed for the rodent Meriones unguiculatus (gerbil), allows flexible stimulus presentations and rewarding as well as on-line data registration. It consists of a spacious housing where the animals have free access to water. Food is supplied exclusively in the y-maze training compartment and serves as a reinforcer in an operant conditioning paradigm.

[Glucocorticoid-induced effect of erythropoietin in haemolytic anaemia with uraemia and red cell enzyme deficiency (author's transl)]. / Glukokortikoid-induzierte Erythropoetinwikung bei urämischen und enzymopenischen Hämolysen.

The effect of 6-methylprednisolone (GCC) was studied on erythropoietin (ESF) levels and on the metabolic functions of erythrocytes (RBC). GCC (U mg/kg/day for 15 days) was administered to 6 patients with the haemolytic-uraemic syndrome (group B) and to 6 patients with non-spherocytic haemolytic anaemia due to hereditary pyruvate kinase enzyme deficiency (group C). 6 healthy persons served as control (group A). The metabolic functions of RBC were investigated by assaying HMPS activity, GSH/GSSG and lactate/pyruvate ratios, relevant glycolytic intermediates, 2,3-DPG, ATP, and key enzymes. A significant increase in ESF was observed in group B patients after GCC therapy, correlating with an improvement in the haemolytic state, and consequent rectification of the secondary disturbances of RBC metabolism. Group C patients already had raised ESF levels before GCC therapy; no further increase occured in response to treatment and no other clinical or haematological change was recorded. Hence, no harmonal influence of GCC on the disturbed RBC metabolic process was detectable in the cases.

[Effects of muscle needle biopsy on parameters of humoral and cellular immunity in pigs]. / Effekte einer Muskelschussbiopsie auf Parameter der humoralen und zellvermittelten Immunität beim Schwein.

The shot biopsy is a common experimental technique for the collection of samples to investigate muscle tissue characteristics or to determine meat quality features in pigs. Its application seems to be also possible in interdisciplinary research projects investigating animal stress, behaviour, and welfare. The present study on 12 group-housed pigs (age: 12 weeks, weight: 29.3 kg) shows the influence of this wound-causing technique on different humoral and cell-mediated parameters of the immune system at 1, 3, 5, and 9 days after biopsy compared to the initially investigated levels before. An enhancement of the blood sedimentation rate and both the IgG and the cellular immune response in vivo (leukocytes, lymphocytes) as well as in vitro (ConA) was observed. Furthermore, there were signs of a secondary wound infection 5 days after biopsies were taken possibly caused by mutual oral manipulation of the animals. We conclude that the muscle shot biopsy technique can be used on group-housed pigs as a method to investigate muscle physiological characteristics. The technique, however, induces immunological reactions which may interfere with stress-induced immune reactions.

Anticipation and frequency of feeding affect heart reactions in domestic pigs.

Measuring heart reactions has become a widely used method for the assessment of emotions. Heart rate and its variability, which can quite easily be noninvasively recorded, reflect the inputs of the sympathetic and parasympathetic branches of the autonomous nervous system. We tested the hypothesis that frequent anticipation of a positive event results in an increased state of welfare in pigs, expressed as positive arousal in anticipation of announced feeding as well as lowered heart rate and augmented heart rate variability during resting periods. We used a controlled paradigm with 3 groups of young domestic pigs (Sus scrofa domestica). We compared frequent acoustic announcement of feed delivery (group 1: 3 feedings between 0730 h and 1030 h plus 3 feedings between 1200 h and 1530 h) with the same number of feedings as in group 1 but without a temporal relation to the sound (group 2) and with a fixed-schedule feeding (group 3: 2 feedings at 0600 h and 1500 h). Specific cardiac and behavioral reactions indicated short-term (1 min) anticipation in the conditioned group. In this group, heart rate increased (P < 0.001) mainly through vagal withdraw and behavior became more active (P < 0.001). Only the conditioned group displayed changing heart rate characteristics during the sound. Pigs in the frequent unpredictable feed group reacted to feed delivery with increased heart rates (P < 0.001), whereas the heart-rate characteristics of pigs with the fixed schedule were unchanged during the sound and while the other 2 treatment groups were feeding. Clear evidence for long-term anticipation (over the course of hours) was not present in the data. Comparisons between the 3 treatment groups suggested that in housing conditions where pigs cannot obtain feed by their actions but must wait for feed delivery, feeding at 2 fixed times would be preferred. Animals in this treatment group presented lower resting heart rates at the end of the experiment than animals in the other 2 groups (P < 0.01). Therefore, merely announcing a positive stimulus without giving control to its access is apparently not suitable for increasing welfare.

Individual calling to the feeding station can reduce agonistic interactions and lesions in group housed sows.

In this study we used a new call feeding station, which enabled sows to learn that they have access to feed only after an individual acoustic signal was given. We tested whether this call feeding station is able to reduce agonistic interactions between sows and whether effects of call feeding can further be improved by enrichment. A total of 85 gestating sows were kept in a dynamic group in a large waiting area (207 m²) equipped with littered laying areas and an outside area. During a control treatment sows were fed in a normal electronic feeding station once a day (NF1-). Before testing the call feeding station sows had been conditioned for an acoustic signal (a trisyllabic "name") and learned that they were allowed to enter the feeding station only after their name was called. In the call feeding station sows were fed either once (CF1-) or twice a day (CF2-). In addition, we tested for effects of further enrichment such as straw in the activity area (CF2+). Agonistic behaviors and number of sows were observed by video in continuous recording from 0600 to 1800 h in an area (4 by 3.25 m) in front of the feeding station in periods of 4 d (NF1- = 7 periods, CF1- = 5 periods, CF2- = 3 periods, and CF2+ = 4 periods) and analyzed using mixed models. During each observation period sows were scored for wounds and body lesions at different body parts. From 0600 to 1100 h the proportion of agonistic interactions was much greater in the feeding mode NF1- compared with CF1- (feeding mode × time of day: P < 0.001) and in CF1- agonistic interactions were on a low level throughout the whole feeding cycle. The feeding frequency and the additional presence of straw in the activity area did not affect the proportion of agonistic interactions (all P > 0.05). The results on the number of sows in front of the feeding station mirrored the findings for agonistic interactions. In NF1- more sows were involved in agonistic interactions compared with CF1- (83.1 ± 12.9% vs. 61.5 ± 19.6%; P = 0.005) but there was no difference between the call feeding station (CF) treatments. The number of severe lesions was greater at the head (P = 0.014) and the flank (P = 0.006) but not at the shoulders (P = 0.057) and the hindquarter (P = 0.426) in NF1- compared with CF1-. The results suggest that signaling the feeding time individually increases the predictability for access to the feeding station and consequently reduces competition between sows.

No apparent effect of an experimental narrow confinement on heart activity and cortisol in domestic pigs.

The effects of continuing spatial restraint were examined in domestic pigs. For this purpose, the animals (German Landrace barrows) were housed individually in metabolic cages (12 animals) and, as controls, in single pens (six animals). In six replications with two experimental animals and one control animal, we collected saliva each morning (0730 h) for the cortisol analysis, recorded the behaviour and the heart beat for 3 h/day (0800 to 0900 h, 1100 to 1200 h, 1400 to 1500 h). Each replication consisted of 5 days of habituation to single housing and 8 experimental days during which the experimental animals lived continuously in the metabolic cages. Weight gain, cortisol, heart rate and heart rate variability were not significantly affected by experimental narrow confinement that only had a significant influence on the behaviours, locomotion and sitting. In conclusion, the experimental animals adapted very fast and did not show physiological indications of chronic stress. However, the absence of apparent stress does not exclude other, subtler, welfare impairments.