[Cortisol concentrations in sheep before, during and after sham foot trimming on a tilt table - the suitability of different matrices]. / Kortisolkonzentrationen bei Schafen vor, während und nach der simulierten Klauenbehandlung im Kippstand ­ Eignung verschiedener Matrices.

INTRODUCTION: Matrices that can be collected non-invasively for quantification of a stress response in sheep have received little attention in the veterinary literature. This study examines the suitability of blood, tears and saliva for determining a stress response in sheep undergoing sham foot trimming on a tilt table. The cortisol concentration of blood, tears and saliva and the concentration of cortisol metabolites in faeces were measured in 13 healthy Meat Merino ewes once a day for six days. Sham foot trimming on a tilt table was used as the stressor and was done during a one-hour period on day 4; cortisol concentrations of blood and tears were measured at 0, 10, 20, 30, 40 and 60 minutes. Cortisol concentrations of blood (maximum at 30 minutes) and tears (maximum at 40 minutes) increased during the procedure and then decreased. There were significant correlations between cortisol concentrations of blood and tears (p = 0,04) during sham foot trimming (area under the curve, 0 to 60 minutes). Over the entire 6-day study period, significant correlations were seen between the cortisol concentrations of blood and tears (r = 0,55; p.

INTRODUCTION: Les marqueurs qui peuvent être collectés de manière non invasive pour quantifier une réponse au stress chez le mouton ont fait l'objet de peu d'attention dans la littérature vétérinaire. Cette étude examine la pertinence du sang, des larmes et de la salive pour déterminer une réponse au stress chez des moutons subissant un parage fictif des pieds sur une table basculante. La concentration de cortisol dans le sang, les larmes et la salive ainsi que la concentration de métabolites de cortisol dans les fèces ont été mesurées chez 13 brebis Meat Merino saines une fois par jour pendant six jours. Le parage fictif des pieds sur une table inclinable a été utilisé comme facteur de stress et a été effectué pendant une période d'une heure le jour 4; les concentrations de cortisol dans le sang et les larmes ont été mesurées à 0, 10, 20, 30, 40 et 60 minutes. Les concentrations de cortisol dans le sang (maximum à 30 minutes) et les larmes (maximum à 40 minutes) ont augmenté au cours de la procédure puis ont diminué. Il y avait des corrélations significatives entre les concentrations de cortisol dans le sang et les larmes (p = 0,04) lors du parage fictif des onglons (aire sous la courbe, 0 à 60 minutes). Sur l'ensemble de la période d'étude de 6 jours, des corrélations significatives ont été observées entre les concentrations de cortisol dans le sang et les larmes (r = 0,55 ; p < 0,001), le sang et la salive (r = 0,53 ; p < 0,001) et les larmes et la salive (r = 0,78 ; p < 0,001). La concentration fécale de métabolites de cortisol était significativement augmentée au jour 5 (p 0,05), mais la concentration de cortisol des autres supports n'a pas changé de manière significative au cours de la période d'étude de 6 jours. Le parage fictif des pieds sur une table basculante a été considéré comme un facteur de stress aigu chez les moutons en raison de l'augmentation des concentrations de cortisol dans le sang, des larmes et de l'augmentation des concentrations de métabolites de cortisol dans les selles. La concentration de cortisol dans les larmes était similaire à celle du sang et, par conséquent, la collecte de larmes représente une alternative viable et non invasive au sang pour les tests de cortisol. Le délai des pics entre la concentration maximale de cortisol dans les larmes et le sang doit être pris en compte lors de l'interprétation des résultats.

The effects of a training program using a phantom to accustom heifers to the automatic milking system.

The introduction of heifers into the automatic milking system (AMS) can be associated with considerable stress for both animals and farm employees, as completely inexperienced heifers initially do not independently enter the unknown milking robot. This study investigated whether training heifers on an AMS phantom provides the possibility of preparing heifers for the following lactation at the AMS. For this purpose, 77 Holstein-Friesian heifers were randomly assigned to one of 2 experimental groups: control (CON) or phantom (PHAN). Four weeks before calving, the PHAN group was given free access to the phantom, which was similar to the actual milking robot, so that they could explore it and be positively conditioned by feeding concentrate in the phantom. The heifers of the CON group had no contact with the phantom or the AMS before the first milking at the AMS. The milking frequency per animal per day was recorded, and the proportion of animals that had to be fetched for milking was determined, to evaluate how the animals accepted the AMS after calving. To assess the stress level of the animals before and after introduction into the AMS, fecal cortisol concentrations and rumination times of the animals were measured. Additionally, lactation performance characteristics (milk yield, milk flow, electrical conductivity of milk, and milk composition) were recorded for 77 animals. The animals trained on the phantom showed a higher milking frequency (DIM 7: 2.70 ± 0.14 visits/d) than the control animals (DIM 7: 2.41 ± 0.14 visits/d) between the 4th and 10th day of lactation. In addition, between d 1 and d 5, the proportion of animals that had to be fetched for milking was lower in PHAN (DIM 1: 35.18 ± 4.16%) than in CON (DIM 1: 48.03 ± 4.46%). The PHAN heifers had unexpectedly high fecal cortisol levels (1 wk prepartum: 43.50 ± 0.93 ng/g of feces), although not considerably elevated compared with CON (1 wk prepartum: 40.76 ± 1.05 ng/g of feces). Training on the phantom had no appreciable influence on rumination time and lactation performance parameters. The increased number of milking visits and the reduced proportion of animals that had to be fetched into the AMS for milking indicate that training on the phantom prepares the animals well for being milked in the AMS. Therefore, training heifers on the phantom offers the possibility to facilitate the start into early lactation for the animals, providing a valuable contribution to improvement of animal welfare.

Seasonal or pathological findings? Morphofunctional characteristics of the equine endometrium during the autumn and spring transition.

The deep anoestrous phase in winter is part of the anovulatory season in mares and is bordered by the autumn and spring transitional periods (ATP/STP). To define an annual time span for effective prognostic biopsy sampling, the aim of this study was to provide a morphofunctional characterization of the endometrium during ATP and STP. To outline both transitional periods, endometrial specimens were taken in September, October and November (n = 76) as well as February, March and April (n = 184) with the requirement of a detailed clinical documentation. Tissue samples were examined histologically with special emphasis on the functional endometrial morphology. Additionally, an immunohistochemical evaluation was performed on selected specimens regarding the expression of oestrogen receptor α, progesterone receptor and Ki67-antigen. An absent to low endometrial activity was ascertained in more than 60% of all specimens from late October onwards, whereas a comparably lacking or low activity in STP was observed until early April. Approximately 30% (ATP) to 22% (STP) of all samples exhibited a predominantly "irregular" endometrial differentiation. During the transitional periods, the clinically evaluated ovarian status (transrectal palpation, transrectal ultrasonography and/or serum progesterone and oestrogen analyses) and the endometrial functional morphology were in accordance with approximately 70% of all cases. The expression of steroid hormone receptors and Ki67-antigen was generally low. Given that endometrial maldifferentiations were frequently found during ATP and STP, its occurence might display a characteristic and physiological feature of the transitional periods. Regarding the functional endometrial morphology, a diagnostic biopsy sampling should therefore be performed between late April and before September.

Radiocarbon constraints on the glacial ocean circulation and its impact on atmospheric CO2.

While the ocean's large-scale overturning circulation is thought to have been significantly different under the climatic conditions of the Last Glacial Maximum (LGM), the exact nature of the glacial circulation and its implications for global carbon cycling continue to be debated. Here we use a global array of ocean-atmosphere radiocarbon disequilibrium estimates to demonstrate a ∼689±53 14C-yr increase in the average residence time of carbon in the deep ocean at the LGM. A predominantly southern-sourced abyssal overturning limb that was more isolated from its shallower northern counterparts is interpreted to have extended from the Southern Ocean, producing a widespread radiocarbon age maximum at mid-depths and depriving the deep ocean of a fast escape route for accumulating respired carbon. While the exact magnitude of the resulting carbon cycle impacts remains to be confirmed, the radiocarbon data suggest an increase in the efficiency of the biological carbon pump that could have accounted for as much as half of the glacial-interglacial CO2 change.

Glycaemic and insulinaemic responses of adult healthy warm-blooded mares following feeding with Jerusalem artichoke meal.

This study aimed to investigate the impact of the supplementation of a pre-biotic compound [Jerusalem artichoke meal (JAM)] on the glycaemic and insulinaemic response in healthy, non-obese warm-blooded horses. Six adult mares [mean body weight (bwt) 529 ± 38.7 kg; body condition score 5.1 ± 0.49/9] were used. In two equal meals per day, the horses received crushed oat grains (1 g starch/kg bwt per day) and meadow hay (2 kg/100 kg bwt per day) which together were likely to meet the energy recommendation for light work (GfE, ). Additionally, they received either 0.15 g fructo-oligosaccharides and inulin (FOS+INU)/kg bwt per day via commercial JAM or maize cob meal without grains as control (CON) in 2 × 3-week periods according to a crossover design. Blood was collected on d21 of the feeding period at different ante- and postprandial (PP) time points (-60, 0, 30, 60, 90, 120, 180, 240 and 300 min), and the plasma glucose and serum insulin levels were determined. Feeding JAM vs. CON did not change the PP peak of glucose or insulin (glucose: 6.3 ± 0.40 vs. 7.0 ± 0.87 mmol/l; insulin: 0.508 ± 0.087 vs. 0.476 ± 0.082 nmol/l) nor did it cause different AUCs until 120 and 300 min PP for glucose and insulin, respectively (AUC120 , glucose: 997 ± 41.6 vs. 1015 ± 41.63 mmol/l per minute, insulin: 49 ± 6.3 vs. 42 ± 6.3 nmol/l per minute; AUC300 , glucose: 1943 ± 142.3 vs. 2115 ± 142.3 mmol/l per minute, insulin: 94 ± 14.8 vs. 106 ± 14.8 nmol/l per minute; p > 0.05). Following JAM vs. CON feeding, glucose and insulin levels declined more rapidly until 240 min PP and tended to be lower (p = 0.053 and p = 0.056, respectively) at this time point. This result might be promising and should further be studied more detailed.

Effects of isoenergetic quantities of a low-starch muesli feed high in fat and fibre vs. oat grains on the glycemic and insulinemic responses and feed intake patterns in sport ponies.

Aim of this study was to compare glycemic and insulinemic responses and feed intake patterns in sport ponies after feeding isoenergetic quantities of low-starch muesli feed high in fat and fibre (FF) or oat grains (OG). Six sport ponies were randomly assigned to one of these two treatment groups for 2 × 3 weeks according to a crossover-design. Ponies received two equal meals/day of either semi-crushed OG (1 g starch/kg bwt*meal-1 ) or an isoenergetic quantity of FF. Hay was also given in two equal meals/day and provided the remaining metabolisable energy up to 1.3-fold maintenance level. On day 21, blood was sampled 1 h after each pony received 0.5 kg hay (0 min). Then, the concentrate was provided and blood sampled 30, 60, 90, 120, 180, 240 and 300 min thereafter. Plasma glucose and serum insulin were analysed, and the areas under the curve (AUC) was calculated 120 and 300 min postprandial (PP). Feed intake patterns were measured in 4 ponies/group via a modified halter. OG was ingested faster than FF (feed intake time; FITDM in min/kg DM: 8.8 ± 1.6 vs. 15.9 ± 1.62, p < 0.05) combined with a higher chewing frequency (p < 0.05). The AUCsgluc120/300, ins120/300 were statistically higher with OG than FF (mmol/L*min-1 : AUCgluc120 : 776 ± 128 vs. 676 ± 80.4; AUCgluc300 : 1811 ± 295.3 vs. 1569 ± 126.3; nmol/L*min-1 : AUCins120 : 38 ± 18 vs. 22 ± 8.1; AUCins300 : 83 ± 39 vs. 35 ± 12; p < 0.05). Plasma glucose tended to decline following the intake of FF, which might be beneficial for equines with reduced glucose tolerance. This, however, requires further investigation. In this study, the ponies consumed OG unexpectedly rapidly. The rate of feed intake was similar to the results previously reported in the literature for warmblood horses.

Endocrine pregnancy monitoring in the two-toed sloth (Choloepus didactylus): "Pregnant or not pregnant".

Progesterone (P4), pregnanediol glucuronide (PdG), estradiol-17ß (E2), and estrone sulfate (E1S) were measured in the feces of four female two-toed sloths (Choloepus didactylus) for early pregnancy diagnosis. For individual feces assignment, the examined female sloths were fed with a turquoise food colorant every second day. Fecal samples were collected one to four times per week, depending on the defecation rate throughout the pregnancies and the postpartum periods. The complete course of pregnancy was subdivided into three 16-week intervals (trimester of pregnancy, TP1-3) and a 5-week post-partum period after birth. Progesterone and PdG concentrations started to increase above luteal phase levels 3 weeks after conception (P = 0.028 and 0.005, respectively). At the beginning of TP1, P4 concentrations averaged 345.0 ± 283.0 ng/g and increased approximately 100- to 300-fold to a peak of 7588.0 ± 6717.0 ng/g over the TP3. Progesterone concentrations were considerably lower than PdG concentrations that started with 3206.0 ± 1500.0 ng/g at TP1 and increased up to 12.8556.0 ± 53.744.0 ng/g until birth. In contrast, mean concentrations of E2 (8.2 ± 2.4-11.7 ± 4.2 ng/g) and E1S (12.2 ± 6.7-22.9 ± 13.0 ng/g) elevated insignificantly and were not suitable for pregnancy detection. All hormones analyzed decreased rapidly within the first weeks after birth. Progesterone and PdG, as well as E2 and E1S, highly significantly correlated (r = 0.602, P < 0.001 and r = 0.497, P < 0.001, respectively) at TP1. During the TP2, only P4 and PdG significantly correlated (TP2: r = 0.661, P < 0.001 and postpartum period: r = 0.616, P = 0.009). In summary, only P4 metabolite concentrations were suitable to determine the status of reproduction in the two-toed sloth. Thereby, PdG was ideally suited to diagnose early pregnancy because it was more sensitive and detected pregnancy 2 weeks earlier than P4.

[Breeding management of black rhinos (Diceros bicornis michaeli) in Magdeburg Zoo]. / Zuchtmanagement bei Spitzmaulnashörnern (Diceros bicornis michaeli) im Zoo Magdeburg.

OBJECTIVE: The African black rhino is an endangered species. In Germany there are only five zoos where this species is kept and breeding has not been successful in all of them. In Magdeburg Zoo the last birth occurred in December 2005, and during the following years, no matings could be observed. During the construction of a new enclosure to enable a more natural mating behaviour, the reproduction status of the rhino cows was evaluated and a hormonal treatment was performed. MATERIAL AND METHODS: Since 2009, faecal samples from two rhino cows (Diceros bicornis michaeli; "Mana", 30 years old, and "Maleika", 17 years old) were collected periodically, and the pregnanediol-glucuronide (PdG) and oestradiol concentrations were determined using enzyme immunoassay and radioimmunoassay, respectively. Following evaluation of the results, both cows were treated for 12 days with Regumate® Equine, a synthetic progesterone, during the period of PdG-dominance. RESULTS: "Mana" accepted the bull 11 days after completion of the hormonal treatment, and in December 2011 gave birth to a healthy calf. "Maleika" had her first ever oestrus 13 days after completion of the medication and also accepted the bull. Thereafter, she had two regular oestrus cycles with normal mating behaviour. Her first calf was born in July 2012. CONCLUSION AND CLINICAL RELEVANCE: The causal treatment of both cows, following a long period of infertility, with synthetic progesterone led to their pregnancy and the birth of healthy calves. The commercial product Regumate® Equine is appropriate to stimulate the sexual cycle in temporarily infertile black rhinos. Attention should be paid to the timing of the medication and the required dose.

[Relationship between natal course and metabolic parameters during the peripartal period in cattle]. / Zusammenhang zwischen Geburtsverlauf und stoffwechselrelevanten Parametern im peripartalen Zeitraum beim Rind.

OBJECTIVE: To evaluate metabolic disorders in high-yielding cattle du- ring the peripartal period, the relationship between the course of the delivery and relevant metabolic parameters (insulin-like growth fac-tor 1 [IGF-1], cortisol and non-esterified fatty acids [NEFA]) were examined. MATERIAL AND METHODS: A total of 28 heavily pregnant cows (13 primiparous and 15 multiparous) with normal and difficult natal courses, respectively, were included in the study. Serum IGF-1, NEFA and cortisol were analysed between day 14 ante partum (a. p.) and day 14 post partum (p. p.), IGF-1 and cortisol were also determined in the newborn calves. RESULTS: An elevated stress situation was present for all heifers primarily at birth, because their serum IGF-1 levels were lower (114 ± 11 vs 158 ± 28 ng/ml) while NEFA and cortisol values were higher (896 ± 76 vs 705 ± 58 µmol/l, p = 0.05; 6.51 ± 1.14 vs 4.67 ± 0.46 ng/ml, p = 0.039, respectively) than those of the cows. During the entire examination period, metabolic disorders could be found in animals with difficult birth, based on significantly lower IGF-1 values (p = 0.003; heifers: p < 0.001), significantly higher NEFA levels (at birth p = 0.05) and higher cortisol values compared to the animals with a normal birth. Similar to their mothers, newborn calves of heifers with dystocia had significantly higher cortisol (p = 0.003) at birth and lower IGF-1 levels (p = 0.002) p. p. compared to calves with a normal natal course. CONCLUSION AND CLINICAL RELEVANCE: Identification and prevention of natal disorders in cattle is very important, especially with regard to animal welfare and economics. In this study, difficult natal courses were associated with altered blood serum concentrations of IGF-1, NEFA and cortisol, which consequently reflect an elevated stress situation during the peripartal period. Furthermore, with regard to these results, it can be concluded that heifers with dystocia are affected by an increased metabolic stress situation mainly during the antepartal period, while multiparous cows suffer from metabolic disorders primarily in the period after birth.

Sexing domestic chicken before hatch: a new method for in ovo gender identification.

Male chicks are an unwanted by-product when producing laying hens. The common practice to kill them directly after they have hatched gives rise to ethical concerns worldwide. The aim of this study was to develop an endocrine method to determine the sex of domestic chicken before hatch. On Days 7 to 10 of incubation, the allantoic fluid from brown layers' eggs (n = 750) was analyzed via enzyme immunoassay for their content of estradiol, estrone sulfate, and testosterone in order to detect gender differences. We successfully established a reliable method for in ovo sex identification on Day 9 of incubation by estrone sulfate measurement in the allantoic fluid. Female embryos displayed significantly higher hormone levels in the allantoic fluid than males (female: median = 0.312 ng/mL; male: median = 0.110 ng/mL; P ≤ 0.001). Our method allows the sexing of domestic chicken at a very early stage of embryonic development, even before the onset of pain perception. The possibility to eliminate eggs containing male embryos on Day 9 of incubation represents a vast improvement compared with culling day-old chicks.

Characterization of the ovarian cycle in the two-toed sloths (Choloepus didactylus): an innovative, reliable, and noninvasive method using fecal hormone analyses.

Little is known about reproductive physiology in the two-toed sloth (Choloepus didactylus). Therefore, the aim of this study was to obtain detailed information about the ovarian cycle. Measurements of reliable gonadal steroids in the feces of this species were undertaken. For this purpose, fecal samples were collected one to three times per week from nonpregnant captive females (n = 2) over a 16-month period. Before assay analysis, the fecal samples were extracted with methanol. Radioimmunoassays and enzyme immunoassays for fecal progesterone, estradiol-17ß, pregnanediol-glucuronide (PdG), and estrone sulfate were tested for their ability to detect the ovarian activity. Using the lowest and highest progesterone values, the ovarian cycle length was comparatively analyzed. The ovarian cycle (n = 26) averaged between 31.4 ± 9.1 days (lowest progesterone) and 32.5 ± 7.5 days (highest progesterone) throughout the whole year. The length of the follicular phase, as indicated by low progesterone levels, was 18.1 ± 4.4 days (range 12-25 days), and the length of the luteal phase, as characterized by elevated progesterone levels, was 13.2 ± 1.8 days (range 11-16 days). In contrast, estradiol-17ß and estrone sulfate were not suitable to detect the cycle due to irregular collection intervals. Fecal progesterone and PdG, as well as estradiol-17ß and estrone sulfate, significantly correlated (r = 0.621, P < 0.01 and r = 0.606, P < 0.01). PdG concentrations (dilution factor (DF) 1:40) were considerably higher than progesterone concentrations (DF 1:10), PdG amounted in the range of 1326.7 ± 320.2 ng/g wet feces (animal S1) and 1373.8 ± 468.3 ng/g wet feces (animal S2) compared with progesterone concentrations in the range of 98.0 ± 17.0 ng/g (S1) and 105.9 ± 30.0 ng/g (S2). The estrone sulfate levels (DF 1:2) were similar to estradiol-17ß (DF 1:22). The mean fecal estradiol-17ß concentrations were 6.7 ± 0.9 ng/g for animal S1 and 7.5 ± 1.6 ng/g for animal S2. In conclusion, the ovarian activity of the two-toed sloth was studied using the noninvasive method by means of the fecal steroid monitoring. Progesterone was the most reliable fecal steroid hormone to determine the duration of the ovarian cycle independent of the weekly defecation rate. The course of progesterone concentrations resulted in a cycle length of 4-5 weeks length in the two-toed sloth. Thus, the reproductive activity of the two-toed sloth does not show any seasonality like the three-toed sloth.

[Interrelationship between peripartal metabolism, performance, morbidity and productive life in a herd of Holstein-Friesian heifers]. / Beziehungen zwischen peripartalem Stoffwechsel, Leistung sowie Morbidität bei Holstein-Friesian-Kalbinnen eines Bestandes und deren Nutzungsdauer.

OBJECTIVE: The aim of this study was to determine the relationship between the heifers' peripartal metabolism, milk yield data and the productive life achieved. Furthermore, the influence of fertility, back fat thickness and morbidity on the productive life was evaluated. MATERIAL AND METHODS: Metabolic data and back fat thickness measurements, determined between 2004 and 2005 in a herd of Holstein-Friesian dairy cows, were analysed retrospectively. Additionally, the age reached by the 207 (at that time) heifers as well as data on culling, fertility, milk yield and morbidity were determined with the help of the herd management system and related to the laboratory findings. RESULTS: Heifers with the shortest productive life of < 12 months had the significantly lowest (p < 0.0001) milk yield. The shortest productive life of < 12 months was associated with the significantly lowest (p < 0.0001) oestradiol concentrations 3 days post partum (p. p.) and the significantly smallest (p < 0.0001) albumin concentrations p. p. Furthermore, heifers with the shortest productive life of < 12 months had the significantly lowest (p < 0.0001) cholesterol concentrations 28 days ante partum (a. p.) and p. p. The data showed that heifers with a short productive life had a tendency for lower back fat thickness, tendencially lower oestradiol concentrations 10 days a. p., a trend to lower fertility (regarding inseminations per animal) and an increased incidence of mastitis, stillbirths and endometritis. CONCLUSION AND CLINICAL RELEVANCE: The lowest albumin and cholesterol concentrations of the heifers with a short productive life arose from an inadequate food intake. Oestradiol concentration also decreased during the state of energy deficiency. The low oestradiol concentrations 10 days a. p. can lead to repeated stillbirths in such heifers, and this increased the number of clinical endometritis. Furthermore, the low milk yield corresponded with an energy deficiency in these heifers. These results were supported by the low back fat thickness of these animals, which consequently led to poor fertility. As a consequence, for a longer productive life, special attention should be paid to heifers during the periparturient period, in particular focusing on their food intake and body condition.

Progesterone, pregnanediol-3-glucuronide, relaxin and oestrone sulphate concentrations in saliva, milk and urine of female alpacas (Vicugna pacos) and their application in pregnancy diagnosis.

The pregnancy-associated hormones, progesterone (P4), pregnanediol-3-glucuronide (PdG), relaxin (RLN) and oestrone sulphate (E1S) in plasma, saliva, milk and urine of alpacas were measured in order to assess their potential use for pregnancy diagnosis. Samples were obtained from 36 female alpacas before mating and at different stages throughout pregnancy (confirmed by ultrasonography). The hormone concentrations were determined using enzyme immunoassays. Milk samples were also tested using a commercial on-farm P4 kit, designed for dairy cattle. Although the concentration of P4 in plasma, milk and urine, and the concentration of PdG in urine were significantly higher in pregnant than in non-pregnant alpacas, there was no difference in the concentrations of P4 or PdG in saliva. The on-farm milk P4 kit showed a sensitivity of 90 per cent for diagnosis of pregnancy and a specificity of 69 per cent for non-pregnancy. The concentration of RLN in plasma increased significantly after the second month, and concentration of E1S in plasma and urine during the last month of pregnancy, whereas, there were no significant differences in RLN or E1S concentrations in saliva and milk between pregnant and non-pregnant alpacas. Values of P4, RLN and E1S in plasma, and PdG and E1S in urine are comparable with the previous reports in alpacas and, therefore, can be confirmed as an indicator for pregnancy. This is the first study to include determination of pregnancy-associated hormones in the saliva and milk of alpacas. However, saliva seems to be unsuitable for pregnancy diagnosis in alpacas, whereas, P4 in milk, as well as PdG and E1S in urine, seem to be adequate tools for diagnosis.

[Psychotherapeutic approaches for patients with somatoform disorders]. / Psychotherapeutische Ansätze für Patienten mit somatoformen Störungen.

Patients with somatoform symptoms are considered to be difficult to treat. Clinical studies on treatment of this condition are underrepresented compared to other mental and psychosomatic disorders. Cognitive-behavioral treatment for patients with somatoform symptoms was found to have a significant effect; additionally, some evidence of recently published findings supports psychodynamic therapy. This report provides information on how to effectively deal with those patients. Furthermore it describes transtherapeutic targets and explains three therapeutic phases: establishing a therapeutic relationship, developing a model of the disorder and establishing coping strategies. As a last point a cognitive-behavioral treatment study, a psychodynamically-oriented study, a group intervention study and a new approach, a combination of cognitive-behavioral therapy and emotion regulation training, are presented.

[Endocrinologic studies of male psittacine birds for the evaluation of their reproductive status]. / Endokrinologische Untersuchung männlicher Papageienvögel zur Beurteilung ihres Reproduktionsstatus.

OBJECTIVE: Evaluating methods of hormone measurement in different specimens of male parrots in order to assess their reproductive status and stress axis. MATERIAL AND METHODS: Cockatiels and rose-ringed parakeets were chosen as psittaciforme representatives and their endocrine profiles were examined. In various pre-experiments, suitable techniques for the determination of testosterone in plasma, saliva and faeces of male parrots were established. Before analysing the samples by enzyme immunoassay, blood and faeces were extracted using diethyl ether, while saliva could be tested without extraction. Based on the excretion of mainly conjugated testosterone metabolites, parrots' faecal samples were also hydrolysed with ß-glucuronidase/arylsulfatase before extraction. In addition, the levels of the stress hormone corticosterone were determined by radioimmunoassay in order to assess possible relationships between stress and the secretion of testosterone. RESULTS: The examined psittacine species displayed different seasonal secretion patterns for both testosterone and corticosterone. Cockatiels had maximum plasma testosterone levels in February, in contrast, rose-ringed parakeets showed highest concentrations in November. As a consequence of cumulative excretion, both species showed much higher faecal than plasma testosterone concentrations. In rose-ringed parakeets, the levels of corticosterone in plasma were exceptionally high compared to the cockatiels. CONCLUSION: According to this study, we have been able to establish suitable methods for testosterone analysis in blood and faeces of cockatiels and rose-ringed parakeets, supporting the assessment of their reproductive status. At present saliva does not appear to be an ideal medium for reliable hormone level measurement, thus further investigations are required concerning this subject. CLINICAL RELEVANCE: By means of process analysis, it will be possible to detect increasing testosterone levels and/or pathological alterations, which could be considered in breeding programmes.

[Bilateral anophthalmia and left-sided orbital tumor : case of an eight-month-old infant]. / Beidseitiger Anophthalmus und linksseitiger Orbitatumor : Fall eines acht Monate alten Säuglings.

An 8-month-old infant from Russia with bilateral anophthalmia presented with an expanding orbital tumor. The tumor was extirpated and the histological examination revealed a non-malignant pseudocystic process with residual neuro-ectodermal structures.

Concentration of platelets and growth factors in canine autologous conditioned plasma.

OBJECTIVES: To report the concentration of blood cells and selected growth factors in canine autologous conditioned plasma (ACP). METHODS: The density of blood cells in whole blood (WB), ACP and standard plasma preparation (SP) of 10 healthy mature dogs was determined. In both ACP and SP, the concentration of insulin-like growth factor-1 (IGF-1), epidermal growth factor, vascular endothelial growth factor, platelet-derived growth factor-AA, platelet-derived growth factor-AB, platelet-derived growth factor-BB, transforming growth factor-ß1 (TGF-ß1), and transforming growth factor-ß2 was measured using the ELISA technique. In another ten dogs, ACP was prepared using an ultra-soft spinning protocol, and again blood cell density was compared to that obtained in WB. RESULTS: The density of platelets in ACP was significantly higher than that in SP (p = 0.0002), but there was not any significant difference between ACP and WB, nor between WB and ACP prepared using softer centrifugations. Interestingly, only for IGF-1, PDGF-BB, and TGF-ß1 could reliable measurements be obtained, showing a significant increase in PDGF-BB and TGF-ß1 concentrations in ACP compared to SP (p = 0.001, p = 0.0028). Regarding IGF-1 content, there was not any significant difference between ACP and SP. CLINICAL SIGNIFICANCE: Canine ACP prepared according to the manufacturer's recommendations, or by using a softer spin does not show the same specifications as human ACP, which shows a doubling in platelet count compared to WB. Even though canine ACP has a similar number of platelets per injected volume and consequently, probably the same amount of injected growth factors than WB, application of canine ACP would not be associated with the proinflammatory potential reported for WB, as it is almost free of erythrocytes and nucleated cells.

Influence of topical dexamethasone applications on insulin, glucose, thyroid hormone and cortisol levels in dogs.

The influence of two topical dexamethasone applications (dermal and ototopical) on plasma insulin, glucose, thyroid hormone and cortisol levels was investigated in beagle dogs. Both treatments significantly decreased basal cortisol values, associated with exaggerated rise in insulin (approximately 50%), together with unchanged serum glucose levels. Dermal dexamethasone quickly decreased plasma thyroxin (T4) levels; whereas dexamethasone in ear drops gradually inhibited time-dependently T4 release (18-50%). Both formulations blunted plasma triiodothyronine (T3) levels but the response induced by dermal dexamethasone was stronger than by dexamethasone ear drops. Upon drug withdrawal, insulin secretion returned to baseline a week after treatment cessation, while cortisol, T4 and T3 levels did not reach baseline values. These results suggest that topical glucocorticoids unexpectedly trigger secondary hypothyroidism with concomitant suppression of hypothalamic-pituitary-adrenal axis but sensitize the endocrine pancreas, thus, their application needs careful evaluation for surprisingly different effects on endocrine stress axis activity.

Investigation of the local expression of the relaxin system in canine mammary tumours.

While mammary tumours are the main reasons of death in bitches, early detection of tumours and metastases is crucial for survival of affected dogs. Invasiveness and angiogenesis, which are important processes of tumour growth and spreading, require connective tissue remodelling. This process is dominantly mediated by matrix metalloproteinases (MMP), which are well known to be positively regulated by relaxin (RLX) in various tissues, including human breast cancer. So far, the presence of RLX and its receptor RXFP-1 as well as their linkage with MMP in canine mammary tumours (CMT) is completely unknown. In the first part of the present study, concentrations of RLX, oestradiol and progesterone from plasma samples of bitches with CMT were compared with clinical and survival data to investigate the predictive value of these hormones. In the second part, the expressions of RLX, RXFP-1 and MMP-2, -9 and -13 were examined by real-time reverse transcriptase polymerase chain reaction (RT-PCR) in 31 CMT samples. Finally, relationships of systemic plasma RLX or locally expressed RLX with expression of MMP in CMT were analyzed for the first time. Comparison of hormone concentrations in 93 bitches in terms of benign or malignant nature of the CMT, lung metastases, recidivation and 12-month survival discovered no significances. The expressions of RLX, RXFP-1 and MMP were independent from plasma RLX, but expressions of local RLX and RXFP-1 showed a strong correlation (p = 0.00004, r = 0.671) as well as RXFP-1 and MMP-2 (p = 0.009, r = 0.463), indicating a possible significant role of the locally produced RLX in CMT pathogenesis as an inducer of connective tissue remodelling.