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.

Monochorial diamniotic dizygotic twins in a German Shepherd Dog: A case report.

CASE REPORT: A 6.5-year-old clinically healthy German Shepherd Dog with regular oestrous cycles of 6 months was presented for pregnancy diagnosis on day 38 after ovulation (p.ov.). Ultrasonography revealed three individual placental sites in progressed resorption and two vital adequately developed foetuses sharing a joint placenta. On days 41 and 48 p.ov., sonographic signs indicated normal development of both foetuses, but on day 52 p.ov., both foetuses were found to be dead. A caesarean section was performed the same day. Examination of the removed uterus confirmed the diagnosis of a "twin" pregnancy with two foetuses sharing the same placental site but separate amniotic membranes. One foetus showed generalized oedema (anasarca). Bacterial culture of swabs taken from inside the placental cavity was negative. At histological examination of the uterus, no signs of inflammation were found. Serum relaxin concentrations (day 38, 41, 48 and 52. p.ov.) were consistent with those of bitches with normal pregnancies. Cytogenetic analysis of the two foetuses revealed dizygotic twins, one male and one female according to SRY-PCR. By genotyping 17 high-polymorphic canine microsatellites, it could be demonstrated that the two foetuses developed from two different oocytes.

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.

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.

Medical treatment improves social behavior in a primate endometriosis model (Callithrix jacchus).

BACKGROUND: As endometriosis is known to be very painful in humans, therapeutic studies should consider pain-related aspects in experimental animals too. In a previous study, we found that common marmosets suffering from endometriosis show remarkable deviations in social behavior and cognitive tasks. METHODS: Female marmosets with endometriosis undergoing medical treatment were tested vs. two control groups (healthy and endometriosis monkeys without treatment) concerning behavioral aspects, cognitive skills, and mobility. RESULTS: The treated monkeys performed significant more social and self-grooming, comfort behavior and were more active than the untreated endometriosis monkeys. The treated marmosets even returned to equal levels as their healthy conspecifics. Cognitive and mobility tests revealed no significant differences. CONCLUSIONS: A beneficial effect of the applied endometriosis medication on behavioral impairments could be obtained. Including such measurements in therapeutic research contributes to aspects of animal welfare and pain-relieving potential of the tested compound.

Parameters of haematology, clinical chemistry and lipid metabolism in the common marmoset and alterations under stress conditions.

Common marmosets are suitable non-human primate models for many human diseases. Standard values for blood parameters are required to evaluate physiological and pathological situations. Two studies were conducted: study I to determine standard values and study II to examine these under changed housing conditions. In study I, all parameters for clinical chemistry were similar in range for both genders with these specifics: male marmosets had significantly higher total and LDL cholesterol levels than females, whereas the mean corpuscular volume and the mean corpuscular haemoglobin were significantly lower than in females. In study II, glucose, lymphocytes and salivary cortisol were significantly lower, and faecal cortisol was increased during the change of housing conditions. In conclusion, standard values for haematology and clinical chemistry for the common marmoset were determined. Further on, parameters that are influenced by relocation stress and its importance for experimental results are described.

Analysis of the bone metabolism by quantitative computer tomography and clinical chemistry in a primate model (Callithrix jacchus).

BACKGROUND: Common marmosets are widely used as experimental primates; however, little is still known about their bone physiology. Therefore, the aim of our study was to analyse body weight, age and bone-specific blood parameters in relation to morphological bone parameters. METHODS: Fifty-eight common marmosets were analysed for blood calcium (Ca), inorganic phosphor (P(i) ), alkaline phosphatase (AP) and 17-ß-estradiol (E2). The examination of bone parameters was undertaken in the lumbar spine by computer tomography. RESULTS: There was a correlation between bone mineral density (BMD) and body weight, trabecular area ratio and polar moment as well as between BMD and AP or Ca (only males), whereas there were no correlations between BMD and age, P(i) or E2 in all analysed genders. CONCLUSIONS: Our data support the assumption that the common marmoset is a reliable primate model to study changes in bone metabolism because of the similarity of our results to humans.

The conclusiveness of less-invasive imaging techniques (computer tomography, X-ray) with regard to their identification of bone diseases in a primate model (Callithrix jacchus).

BACKGROUND: Although common marmosets seem to be appropriate animal models to examine bone diseases, no data about the conclusiveness of less-invasive techniques are available. Therefore, the aim was to combine different techniques to analyse changes in bone metabolism of common marmosets with bone diseases. METHODS: Five monkeys were examined by X-ray, computer tomography (CT), histology and immunohistochemistry (IHC). RESULTS: Monkeys with lowest bone mineral density (BMD) showed increased bone marrow, decreased cancellous bone and decreased contrast in X-ray. Highest alkaline phosphatase (AP)-levels were detected in bones with low elastic modulus. Expression of osteopontin (OPN), osteocalcin (OC) and runt-related transcriptions factor 2 (RUNX 2) was detected in bones with high modulus. No expression was present in bones with lower modulus. Collagen type I and V were found in every bone. CONCLUSIONS: In conclusion, CT, X-ray and AP are useful techniques to detect bone diseases in common marmosets. These observations could be confirmed by IHC.

Serum progesterone in pregnant bitches supplemented with progestin because of expected or suspected luteal insufficiency.

Progesterone profiles of individual bitches may vary considerably both between and within individuals during pregnancy and non-pregnancy. Suspected luteal deficiency is commonly purported but is difficult to evaluate in clinical cases when progesterone is supplemented because this masks the underlying hormone changes. Therefore, in this study, suspected cases of luteal deficiency (six pregnancies from five bitches) were supplemented with oral medroxyprogesterone acetate (MPA), allowing measurement of endogenous progesterone using conventional assay. MPA (0.1 mg/kg) treatment commenced between days 30 and 36 after estimated ovulation and was continued for 18-28 days. Endogenous progesterone was measured throughout treatment, and blood was additionally analysed for prolactin (PRL) and relaxin (RLN) as well as MPA. The latter revealed delayed MPA clearance in two bitches, in which Caesarean operation had to be performed because of a low foetal heart rate. In two cases with confirmed basal concentrations of both P(4) and MPA at term, spontaneous parturition occurred. Low endogenous progesterone during pregnancy was not apparent in three bitches that had previously had a short inter-oestrous interval of which two had previously had confirmed short luteal phase. However, in the remaining two cases, there had been previous pregnancy failure, but in only one of these, a premature decrease in endogenous progesterone to <2 ng/ml was detected. The latter had also low concentrations of PRL and RLN. The results of this preliminary clinical study suggest that abnormal progesterone profiles in pregnancy may be uncommon in bitches even when there has been previously documented short inter-oestrous interval. However, luteal deficiency may be suspected in bitches with a history of repeated pregnancy failure or abortion. MPA supplementation appears to be efficacious for management of suspected luteal deficiency and verification of the ovarian dysfunction, but care should be taken regarding the timing of MPA withdrawal and prolongation of pregnancy because of delayed elimination of MPA from blood circulation.

Testicular steroids, prolactin, relaxin and prostate gland markers in peripheral blood and seminal plasma of normal dogs and dogs with prostatic hyperplasia.

Concentrations of 17ß-oestradiol (E(2) ), testosterone (T), 5α-dihydrotestosterone, prolactin (PRL) and relaxin (RLN) were determined in peripheral blood serum or plasma and prostatic secretion of 77 physically healthy intact male dogs (19 Rhodesian Ridgebacks/RR, 58 dogs of other breeds, 1-9 years of age). Furthermore, the concentrations of acid phosphatase in prostatic secretion and canine prostate-specific esterase (CPSE) were measured in blood plasma. All dogs were submitted to a complete breeding soundness examination, including B-mode sonography. Prostatic volume was larger, and blood plasma levels of CPSE were higher in ageing dogs and in dogs with benign prostatic hyperplasia (BPH) compared with young dogs and dogs with normal prostate. Furthermore, a higher E(2) /T ratio was found in dogs with BPH. Despite missing significant differences in PRL concentrations, the slight increases in PRL concentrations in the prostatic secretion observed both with increasing age and in dogs with BPH and the observed correlations between concentrations of PRL and testicular steroids may possibly indicate a role of PRL in the pathogenesis of canine BPH. Serum RLN concentrations were at similar level in all dogs. Regarding breed differences, an appreciably larger prostatic volume and higher concentration of CPSE were verified in RR than in other pure-bred dogs, confirming our suspicion of a premature enlargement of the prostate gland, which may result from a genetic disposition for BPH in this breed.

Behavioral tests as indicator for pain and distress in a primate endometriosis model.

BACKGROUND: As common marmosets (Callithrix jacchus) are frequently used experimental animals, sensitive test systems are needed to evaluate impairment and pain caused by procedures and diseases. METHODS: A diurnal profile of healthy animals was obtained by videotaping. Differences in social behavior and cognitive skills between marmosets with established endometriosis and healthy monkeys were investigated using the videotaping, the Wisconsin General Test Apparatus (WGTA), and a food tree. RESULTS: The marmosets showed a mostly trimodal course of activity. Social grooming and activity were significantly decreased in animals with endometriosis; furthermore, the diseased monkeys habituated significantly worse to the cognitive test settings. The food tree experiments offered no differences between diseased and control animals. CONCLUSION: The videotaping and the WGTA are suitable methods to detect disease-related impairments in common marmosets, which is essential for the refinement of experiments.

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.

[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.

[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.

Expression of ABC-transport proteins in canine mammary cancer: consequences for chemotherapy.

Intrinsic or acquired drug resistance is a major barrier for chemotherapy of cancer. Importantly, the presence of ATP-binding cassette, ABC-transport proteins in tumour cells circumvents an intracellular accumulation of chemotherapeutic drugs. In this study, 103 canine mammary tumour probes were investigated for mRNA expression of seven ABC-transporters by RT-PCR. All tumour samples expressed multidrug resistance-associated protein 1 (MRP1) and breast cancer resistance protein (BCRP). MRP7 was detected in 97.1% of tumour probes, MRP3 in 96.1%, Pgp in 92.2%, MRP5 in 85.4% and MRP6 in 64.1%. More of the half of tumour samples (56.1%) expressed all of the examined ABC-transport proteins. Approximately one-third of the tumour samples (32.7%) were lacking in one transporter and only 11.2% possessed from three to five transporters. The canine transporter cBCRP was functionally analysed in stable transfected Madin-Darby canine kidney-II cells using an MTT viability test. cBCRP transfected cells showed a 5.4-fold resistance to 10 microm doxorubicin. Cell survival in the presence of methotrexate was not affected by cBCRP. In conclusion, absence of efficiency of chemotherapy of canine mammary cancer can be caused by expression of seven various ABC-transport proteins. Because cBCRP is expressed in all examined tumour probes and induces resistance to doxorubicin, the application of doxorubicin for treatment of canine mammary is inappropriate.

Comparison of selected endocrine parameters during luteal phase and pregnancy in German Shepherd dogs and Beagles.

Concentrations of progesterone, prolactin and relaxin in serum at predetermined intervals after ovulation (day 0) in non-pregnant and pregnant normocyclic Beagles were assayed and results compared with those observed in German Shepherd dogs (GSD) in a previous study. The goal was to determine possible reproductive hormone specificities related to the GSD breed. Furthermore, the effects of medroxyprogesterone acetate (MPA)-treatment in non-pregnant Beagles and of progesterone supplementation in pregnant Beagles on the hormone concentrations were examined. Mean concentrations of progesterone and prolactin were not different in the non-pregnant Beagles compared with those seen in non-pregnant GSD, except at days 50-60, when progesterone concentrations were found to be higher in Beagles (p < 0.05). Mean progesterone concentrations in pregnant Beagles at days 50-60 after ovulation (day 0) were higher (p < 0.05) than in GSD at that time, but not at earlier time periods. Prolactin concentrations were higher (p < 0.05) in Beagles throughout pregnancy compared with those in the GSD. Mean relaxin concentrations were numerically but not significantly lower in GSD than in Beagles throughout pregnancy. A 10-day oral MPA treatment did not affect progesterone or prolactin secretion in normocyclic non-pregnant Beagles. Medroxyprogesterone acetate serum concentrations were approximately 3.9 ng/ml during treatment and decreased to 0.42 and 0.021 ng/ml within 5 and 15 days after end of treatment, respectively. Intramuscular progesterone supplementation from days 30 to 40 in pregnant Beagles resulted in higher concentrations of progesterone in the 36- to 45-day time periods; prolactin and relaxin concentrations were not significantly affected during or after treatment compared with administration of placebo. The results suggest a tendency towards deficient luteal function in the short-cycle GSD bitches previously studied, which in pregnancy may reflect the observed decreased prolactin concentrations; the possibility that GSD relaxin secretion is deficiency required needs further study. As oral treatment with MPA did not affect progesterone and prolactin release, it may be useful for studying luteal function in pregnant bitches with suspected hypoluteoidism.

Morphological examination of the corpora lutea from pregnant bitches treated with different abortifacient regimes.

Different abortifacient regimes in dogs were analysed for their effect on the pregnancy corpora lutea (CL), namely, prostaglandin F2a analogue cloprostenol (CLO) combined with dopamine agonist cabergoline (CAB), or progesterone (P4) receptor antagonist aglepristone (AGL). Ovaries were collected after 6-10 days of treatment during first trimester. The CL of the control-group showed strong expression of relaxin (RLX), its receptor RXFP1 and enzymes of steroid biosynthesis (HSD) with high peripheral P4-levels. Whereas RXL, RXFP1 and HSD were lowest expressed in the CLO/CAB-group with a massive degeneration of CL and their blood vessels combined with low peripheral P4-level. The AGL-group showed less extensive CL degeneration and more intensive staining of the examined factors than CLO/CAB. In summary, all examined factors are associated with normal luteal function and are useful tools to stage luteolysis. Although both treatments have the same abortive action, their sequence of events on the CL is different.

Influence of progesterone withdrawal on pregnancy-related parameters during post-implantation early pregnancy loss.

Pharmacologically-induced luteolysis or treatment with an antiprogestin in early post-implantation pregnancy in dogs results in asynchronous death and resorption of conceptuses, indicating variable rates of response of individual conceptuses towards progesterone deficiency. This variability also seems to occur in bitches showing pregnancy failure in response to spontaneous luteal deficiency. In a total of 10 beagle pregnancies (two consecutive pregnancies of five bitches), abortifacient treatments beginning on day 24 after ovulation (ov) involved either administration of a progestin antagonist (total of six pregnancies, in three bitches) or a luteolytic regimen of prostaglandin F(2alpha)-analogue together with a dopamine agonist (total of four pregnancies, in two bitches). The outcomes were evaluated in relation to four control pregnancies in two bitches by assay of serum progesterone, prolactin and relaxin at selected time points or within selected time periods, by ultrasound of conceptuses including measurement of uterine blood flow, and parameters of the blood fibrinolytic system including plasma fibrinogen and plasminogen. The process of embryonic death and conceptus resorption was variable in onset and duration both in bitches that received the progesterone antagonist aglepristone (AGLE) and in those under the luteolytic treatment (cloprostenol combined with cabergoline). Pregnancy termination (death of all embryos or foetuses, respectively) occurred as early as day 29 and as late as day 41 after ov in AGLE-treated bitches, and not earlier than day 37 after ov in luteolytic-treatment bitches. Impending embryonic death was not predicted by changes in relaxin concentration, parameters of the fibrinolytic system, or in the perfusion of small uteroplacental vessels.

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.