Effects of long-term dietary supplementation with conjugated linoleic acid on bovine oocyte lipid profile.

Nutritional and environmental conditions around conception and during early embryonic development may have significant effects on health and well-being in adult life. Here, a bovine heifer model was used to investigate the effects of rumen-protected fat supplementation on oocyte quality and embryo development. Holstein-Friesian heifers (n=84) received a dietary supplement consisting of rumen-protected conjugated linoleic acid (CLA) or stearic acid (SA), each on top of an isocaloric basic diet. Oocytes were collected via ultrasound-guided follicular aspiration and subjected to in vitro maturation followed by either desorption electrospray ionisation mass spectrometry (DESI-MS) for lipid profiling of individual oocytes or in vitro fertilisation and embryo culture. The type of supplement significantly affected lipid profiles of in vitro-matured oocytes. Palmitic acid and plasmalogen species were more abundant in the mass spectra of in vitro-matured oocytes after rumen-protected SA supplementation when compared with those collected from animals supplemented with CLA. Lipid concentrations in blood and follicular fluid were significantly affected by both supplements. Results show that rumen-protected fatty-acid supplementation affects oocyte lipid content and may pave the way for the establishment of a large-animal model for studies towards a better understanding of reproductive disorders associated with nutritional impairments.

The effect of metritis and subclinical hypocalcemia on uterine involution in dairy cows evaluated by sonomicrometry.

The objective of this study was to examine the effects of metritis and subclinical hypocalcemia on reduction of uterine size in dairy cows using ultrasonography and sonomicrometry. Four piezoelectric crystals were implanted via laparotomy into the myometrium of the pregnant uterine horn of 12 pluriparous Holstein Friesian cows 3 weeks before the calculated calving date. Sonometric measurements were conducted daily from 2 days before parturition (= Day 0) until Day 14 after calving and then every other day until Day 28. Distances between adjacent crystals were expressed in relation to reference values obtained before calving. The diameter of the formerly pregnant uterine horn was measured using transrectal B-Mode sonography starting on Day 10. Cows were retrospectively divided into the following groups: cows without metritis (M-; n = 7), cows with metritis (M+; n = 5), cows with normocalcemia (SH-; Ca > 2.0 mmol/l on Days 1 to 3; n = 5) and cows with subclinical hypocalcemia (SH+; Ca < 2.0 mmol/l in at least one sample between Days 1 and 3; n = 7). Metritis did not affect (P > 0.05) sonometric measurements, but the diameter of the formerly pregnant horn was larger (P ≤ 0.05) between Days 15 and 21 in M+ cows than in M‒ cows. Reduction in uterine length in hypocalcemic cows was delayed (P ≤ 0.05) between Days 8 and 21 compared with normocalcemic cows, but the uterine horn diameter was not related to calcium status. In conclusion, both diseases affected reduction of uterine size until Day 28. Cows with metritis had a larger uterine diameter, possibly attributable to accumulation of lochia, and cows with subclinical hypocalcemia had delayed reduction of uterine length, presumably related to reduction of myometrial contractility.

The effect of metritis on luteal function in dairy cows.

BACKGROUND: Disturbed uterine involution impairs ovarian function in the first weeks after calving. This study analyzed the long-term effect of metritis on luteal function of 47 lactating Holstein-Friesian cows during the first four postpartum estrous cycles. Cows with abnormal uterine enlargement and malodorous lochia were classified as having metritis (group M, n = 18), and all others were considered healthy (group H, n = 29). Luteal size was measured once between days 9 and 13 of the first (group H, n = 11; group M, n = 12), second (group H, n = 23; group M, n = 18) and fourth (group H, n = 11; group M, n = 7) postpartum luteal phases. Serum progesterone concentration was measured at the same time. Sixteen cows (group H, n = 9; group M, n = 7) underwent transvaginal luteal biopsy for gene expression analysis of steroidogenic regulatory proteins during the second and fourth cycles. Cows with persistence of the corpus luteum (CL) underwent determination of luteal size, luteal biopsy and serum progesterone measurement once between days 29 and 33, followed by prostaglandin treatment to induce luteolysis. The same procedures were repeated once between days 9 and 13 of the induced cycle. RESULTS: The cows in group M had smaller first-cycle CLs than the cows in group H (p = 0.04), but progesterone concentrations did not differ between groups. Luteal size, progesterone concentration and gene expression did not differ between the two groups during the second and fourth cycles. Compared with healthy cows (10%), there was a trend (p = 0.07) toward a higher prevalence of persistent CLs in cows with metritis (33%). Persistent CLs were limited to the first cycle. Persistent CLs and the induced cyclic CLs did not differ with regard to the variables investigated. CONCLUSIONS: An effect of metritis on luteal activity was apparent in the first postpartum estrous cycle. However, after the first postpartum cycle, no differences occurred in analyzed parameters between metritis and control cows. Therefore, a metritis is able to impair luteal activity transiently, but does not seem to have a long-term effect on luteal function.

Characterization of recombinant human and bovine thyroid-stimulating hormone preparations by mass spectrometry and determination of their endotoxin content.

BACKGROUND: The TSH stimulation test to confirm canine hypothyroidism is commonly performed using a recombinant human TSH (rhTSH), as up to date, canine TSH is not yet commercially available. Limiting factors for the use of rhTSH are its high costs and occasional difficulties in product availability. Less expensive bovine TSH preparations (bTSH) purified from bovine pituitary glands are readily commercially available. The aim of this study was to evaluate two different bTSH products as alternative to rhTSH using mass spectrometry. RESULTS: More than 50 proteins, including other pituitary hormones, bovine albumin, hemoglobin, and tissue proteins were identified in the bTSH preparations. In contrast, rhTSH proved to be a highly pure product. Significantly higher endotoxin levels could be detected in all bTSH products compared to the rhTSH. CONCLUSIONS: Both bTSH products are crude mixtures and therefore not an acceptable alternative to rhTSH. Their use should be discouraged to prevent unintended side effects.

The anabolic steroid methandienone targets the hypothalamic-pituitary-testicular axis and myostatin signaling in a rat training model.

There is increasing evidence that the biological activity of myostatin (MSTN), a negative regulator of muscle growth, is affected by training but also anabolic steroids. In this study, we analyzed the effects of the frequently abused anabolic steroid methandienone (Md) on the hypothalamic-pituitary-testicular axis and androgen-sensitive tissues in intact rats performing a treadmill training to simulate the situation of abusing athletes. The anabolic effects were correlated with the expression of members of the MSTN signaling cascade. Md treatment resulted in a significant stimulation of anabolic activity of the levator ani muscle, which was further increased by training, while prostate and seminal vesicle weights decreased in conformance with hormone concentrations of LH and testosterone. In gastrocnemius muscle, mRNA expression of genes of the MSTN signaling cascade (MSTN, Smad7 and MyoD) was reduced by training but not after Md treatment, in soleus muscle MSTN and its inhibitors, follistatin (FLST) and Smad-7 were only affected after training in combination with Md treatment. In summary, our data demonstrate that Md treatment of intact rats results in anabolic effects which are enhanced in combination with physical activity. Interestingly, the anabolic activity on the levator ani was increased in combination with training, although the levator ani muscle was not specifically stimulated by our training protocol. In the m. gastrocnemius and soleus, the anabolic effects correlate with changes in the expression patterns of genes involved in MSTN signaling. Our data provide evidence that the decrease in the weight of androgen-sensitive sexual glands, observed after Md treatment, is caused by a suppression of endogenous testosterone synthesis. These observations provide new insights into the molecular mechanisms of the interaction between anabolic steroids, training and MSTN signaling during skeletal muscle adaptation.

Comparison of isoflurane inhalation anaesthesia, injection anaesthesia and high volume caudal epidural anaesthesia for umbilical surgery in calves; metabolic, endocrine and cardiopulmonary effects.

OBJECTIVE: To compare three anaesthetic protocols for umbilical surgery in calves regarding adequacy of analgesia, and cardiopulmonary and hormonal responses. STUDY DESIGN: Prospective, randomised experimental study. ANIMALS: Thirty healthy German Holstein calves (7 female, 23 male) aged 45.9 ± 6.4 days. METHODS: All calves underwent umbilical surgery in dorsal recumbency. The anaesthetic protocols were as follows: group INH (n = 10), induction 0.1 mg kg(-1) xylazine IM and 2.0 mg kg(-1) ketamine IV, maintenance isoflurane in oxygen; Group INJ (n = 10), induction 0.2 mg kg(-1) xylazine IM and 5.0 mg kg(-1) ketamine IV, maintenance 2.5 mg kg(-1) ketamine IV every 15 minutes or as required; group EPI (n = 10), high volume caudal epidural anaesthesia with 0.2 mg kg(-1) xylazine diluted to 0.6 mL kg(-1) with procaine 2%. All calves received peri-umbilical infiltration of procaine and pre-operative IV flunixin (2.2 mg kg(-1) ). Cardiopulmonary variables were measured at preset intervals for up to 2 hours after surgery. The endocrine stress response was determined. Intra-operative nociception was assessed using a VAS scale. Data were compared between groups using appropriate statistical tests. A value of p < 0.05 was considered significant. RESULTS: All three protocols provided adequate anaesthesia for surgery although, as judged by the VAS scale, intra-operative response was greatest with INJ. Lowest mean cortisol levels during surgery occurred in EPI. Heart rate and cardiac output did not differ between groups, but mean arterial blood pressure, systemic vascular resistance, and partial pressure of carbon dioxide were higher and arterial pH lower in groups INH and INJ than in Group EPI. Group INJ became hypoxaemic and had a significantly greater vascular shunt than did the other groups. CONCLUSION AND CLINICAL RELEVANCE: Groups INH and EPI both proved acceptable protocols for calves undergoing umbilical surgery, whilst INJ resulted in variable anti-nociception and in hypoxaemia. High volume caudal epidural anaesthesia provides a practical inexpensive method of anaesthesia for umbilical surgery.

Assessment of the effects of adjunctive gabapentin on postoperative pain after intervertebral disc surgery in dogs.

OBJECTIVE: To assess the effect of adjunctive gabapentin (GBP) on pain after thoracolumbar intervertebral disc surgery in dogs. STUDY DESIGN: Prospective, randomized, controlled, clinical, 'blinded' trial. ANIMALS: Sixty-three client owned dogs undergoing hemilaminectomy METHODS: Dogs were assigned to two treatment groups. The GBP group received gabapentin 10 mg kg(-1) orally every 12 hours starting before anaesthesia; the placebo (P) group received empty gelatin capsules. Background analgesia was initiated with intravenous levomethadone 0.6 mg kg(-1) (as the combination 'L-Polamivet) at anaesthesia induction, followed by a fentanyl patch and levomethadone 0.2 mg kg(-1) subcutaneously every 8 hours for 24 hours. Pain was assessed by the short form of the Glasgow Composite Measure Pain Score (CMPS-SF) without the gait category, and by a Visual Analogue Scale (VAS). Serum GBP concentrations and cortisol concentrations were measured. Statistical analyses utilized chi square test, Kolmogorov-Smirnov test, two-way analysis of variances for repeated measurements, Wilcoxon test and Friedmann test as relevant. Correlations were tested by Spearman's and Pearson's correlation coefficient. p < 0.05 was considered significant. RESULTS: Median CMPS-SF was lower in group GBP than in group P on days 0.5, 1, 4 and 5. However, CMPS-SF and VAS were not significantly different between groups. Both pain scores decreased significantly over time. Cortisol concentrations were not significantly different between groups. Minimum serum concentrations of GBP fell below the detection limit of 1 µg mL(-1) in 6 of 29 and 7 of 28 dogs at 24 and 72 hours, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: 10 mg kg(-1) GBP orally twice a day did not result in a detectable reduction in pain behaviour compared to background opioid analgesia alone, although a trend to lower pain levels (p < 0.1) was present. Further studies are needed to determine if this is related to effective background analgesia or an ineffective dose of GBP.

Modulation of electrolyte homeostasis by dietary nitrogen intake in growing goats.

In goats, the combination of dietary N and Ca reduction caused hypocalcaemia and further changes in Ca homeostasis. The aim of the present study was to characterise the effects of dietary N reduction under normocalcaemia on mineral and bone metabolism in young goats. Young male goats of the Saanen breed were fed a diet reduced in N (8 %) for about 7 weeks (ten animals per group) and were compared with goats fed with an adequate N (14 %) diet. When N intake was reduced in young goats, plasma urea concentrations as well as renal elimination of urea were reduced. This was inversely related to creatinine in plasma and urine, which increased during a dietary N reduction as a function of reduced renal activity to save urea during N scarcity. During this decrease in renal function, associated with declined insulin-like growth factor 1 concentrations, a reduction in calcidiol and calcitriol concentrations could be observed. Meanwhile, carboxyterminal cross-linked telopeptide of type I collagen values and activity of total alkaline phosphatase were both elevated, indicating some bone remodelling processes taking place during a reduced N diet in young goats. The concentrations of inorganic phosphate (Pi) and total Ca were changed in several body fluids, indicating that Pi and Ca homeostasis was perturbed in goats fed a reduced N diet. Therefore, more research is needed to find the balance between reduction of environmental N pollution by reducing dietary N in ruminant feeding and maintaining the animal's health.

Comparison of commercial ELISA blood tests for early pregnancy detection in dairy cows.

The objective of the present study was to compare two commercially available blood-based pregnancy tests, namely BioPRYN, an ELISA for pregnancy-specific protein B (PSPB), and an ELISA for pregnancy-associated glycoprotein (PAG), for early pregnancy diagnosis in dairy cattle using transrectal ultrasonography as a gold standard. Transrectal ultrasonography was conducted 26-58 days after artificial insemination (AI) in 197 cattle from 19 farms. Concurrently, a blood sample was collected for determination of serum PSPB and PAG. Transrectal palpation was performed approximately 120 days after AI to verify that pregnancy was maintained. For PSPB and PAG, there were no significant differences (P>0.05) in sensitivity (98.0 and 97.8%), specificity (97.1 and 91.2%), positive predictive values (99.3 and 97.8%), negative predictive values (91.9 and 91.2%) and accuracy (97.8 and 96.4%). In conclusion, the two blood pregnancy assays were equally efficacious and were highly accurate (based on transrectal ultrasonography as the gold standard).

Kindling as a model of temporal lobe epilepsy induces bilateral changes in spontaneous striatal activity.

Basal ganglia are engaged in seizure propagation, control of seizures, and in epilepsy-induced neuroplasticity. Here, we tested the hypothesis that previously observed histological and neurochemical changes in the striatum of amygdala-kindled rats as a model of temporal lobe epilepsy are reflected in alterations of spontaneous striatal firing rates and patterns. Because experimental histological and clinical imaging studies indicated a bilateral involvement of the striatum in epilepsy-induced neuroplasticity, in vivo single-unit recordings were done bilaterally 1 day after a kindled seizure in rats kindled via the right amygdala. Compared to control animals, we observed (1) an increased irregularity of firing of neurons classified as striatal projection neurons and located in the anterior striatum ipsilateral to the kindling side and (2) an increased spontaneous activity of neurons classified as striatal projection neurons and located in the anterior striatum contralateral to the kindling side. These hyperactive neurons were located within the dorsolateral (sensorimotor) subregion of the striatum. The present study represents the first evidence of kindling-induced bilateral changes in electrophysiological properties of striatal neurons and demonstrates that the striatum is strongly affected by the functional reorganization of neurocircuits associated with kindling. The changes are probably caused by a combination of several factors including disturbed bilateral limbic and neocortical input as well as disturbed intrastriatal GABAergic function. The changes reflect a pathophysiological state predisposing the brain to epileptic discharge propagation or else (contralateral striatum) could represent a compensatory network of inhibitory circuits activated to prevent the propagation of seizure activity. The findings are relevant for a better understanding of kindling-induced network changes and might provide new targets for therapeutic manipulations in epilepsies.

Influence of testosterone on the docility of German Simmental and Charolais heifers during behavior tests.

In beef cows, docility is important for animal welfare reasons, the farmer's and veterinarian's safety as well as for economic reasons. The hormone testosterone is often mentioned in the context with aggressions and docility. The present study tested the hypothesis that higher testosterone levels in beef cows are associated with more aggressive behavior and lower docility during behavior tests. German Simmental (beef) (n=21) and Charolais heifers (n=20) were subjected with one repetition to a chute test as well as a separation and restraint test. Heart rate, thermal images of the eye, saliva cortisol as well as saliva testosterone were collected along with behavioral parameters during the tests. For all physiological and most of the behavioral parameters significant breed differences were detected. Charolais heifers (0.10 ± 0.01 ng/ml saliva) had considerably higher testosterone levels compared with Simmental heifers (0.04 ± 0.01 ng/ml; P=0.0001). Interestingly, the Charolais heifers expressed a higher docility, so across breeds higher levels of testosterone are associated with higher docility. However, all the parameters presumably linked to stress (heart rate, eye temperature, cortisol levels) generally showed stronger correlations to behavior traits than testosterone (e.g., trait "time in corner" - testosterone: r=0.25 vs. cortisol: r=-0.43 (Fisher's r-to-z: P=0.0985); eye temperature: r=-0.65; (P=0.0005); heart rate: r=-0.60 (P=0.003)). Eye temperature correlated with both heart rate (r=0.68) and cortisol levels (r=0.62; both P<0.0001). Therefore, eye temperature more so than testosterone appears to be a suitable indicator of stress, and as such also an, albeit incomplete, indicator of docility in cattle.

[Pregnancy diagnosis by detection of pregnancy-associated glycopoteins in milk: evaluation of a commercial available ELISA]. / Trächtigkeitsdiagnostik per Milchprobe: Evaluation des Nachweises von trächtig- keitsassoziierten Glykoproteinen mittels kommerziellem ELISA.

In order to increase fertility in modern dairy farming, the interval between first insemination and conception should be as short as possible. Therefore, one approach is to diagnose non pregnant animals early to re-inseminate them as soon as possible. Commercial milk pregnancy assays are available to detect throphoblast derived bovine pregnancy-associated glycoproteins (bPAG) in milk. The aim of the present study was first to evaluate pre-analytical factors interfering with the correct detection of bPAG in milk. To achieve this aim, the stability of the bPAG was tested after repeated freezing and thawing cycles, as well as after a storage of seven days at room temperature and at 37°C. Secondly, the diagnostic performance of a commercially available PAG-ELISA was evaluated for pregnancy diagnosis between 28 to 60 days after artificial insemination by comparing the results with transrectal ultrasonography in a field study (n = 291 cows). After one freezing and thawing procedure the optical density (OD) increased and afterwards stayed stable. The storage of milk samples at room temperature had no effect on the OD, but after five days of storage at 37 °C the OD dropped sharply. It is therefore recommended to add an appropriate microbicidal preservative for shipment and storage of milk samples for bPAG analysis . The results of the field study showed that PAG-ELISA results and ultrasonography results agreed in 95.7% of the cases. Nine samples were tested as "false negative"and five as "false positive", however a re-check revealed that all "false positive"and "false negative" tested cows suffered from embryonic/fetal mortality. In conclusion, the pregnancy diagnostic by bPAG detection in milk samples is an accurate method to diagnose early pregnancy in lactating cows.

Validity of physiological biomarkers for maternal behavior in cows--a comparison of beef and dairy cattle.

The objectives of the present study were to evaluate the suitability of potential biomarkers for maternal ability in cattle, and in addition to test the hypothesis that dairy cows have a less pronounced motherliness than beef cows. Therefore, maternal behavior of 20 Simmental beef-type (S) and 20 German Black Pied (dairy-type) Cattle (BP) was assessed on the 2nd and again on the 3rd day of the calf's life. Measurements included the frequency of interactions between cow and calf, the cow's willingness to defend her calf, the overall maternal behavior, saliva cortisol, saliva oxytocin, heart rate, and thermal images of the eye (ET). Mixed model analysis revealed that BP had significantly (P<0.05) higher oxytocin (88.6±9.2 vs. 62.8±9.2 pg/ml saliva) and cortisol (1.3±0.1 vs. 1.0±0.1 ng/ml saliva) levels, but lower heart rates (80.0±2.0 vs. 95.8±2.0bpm) than S cows. Simmental (beef) cows showed more defensive behavior (3.5±0.2 vs. 2.7±0.2 scores), but fewer total interactions between cow and calf (8.1±1.4 vs. 13.8±1.4), compared to BP (dairy). However, with the exception of heart rate and overall maternal behavior, breed differences tended to diminish from the 2nd to the 3rd day of the calf's life. Repeatabilities ranged from 9±23% (ET) to 77±7% (maternal behavior measured on a visual analogue scale), and correlations between physiological parameters and behavior differed between breeds and were generally at a low level. In conclusion, beef cows do not seem to be per se more maternal compared to dairy cows, and the assessed parameters are of limited use as biomarkers for maternal behavior.

Testicular yolk sac tumor and impaired spermatogenesis in a Holstein Friesian calf.

Yolk sac tumors are testicular germ-cell tumors of the non-seminoma type. In cattle, this neoplasm is very rare and to date has only been described three times. In human males, it usually occurs in infants and children. Immunohistochemistry provides solid criteria for diagnostics. Especially present pathognomonic Schiller-Duval bodies are helpful for identification. In this report, a 32-day-old Holstein Friesian calf presented with a highly enlarged right testis. Sonographic examination was performed and blood samples were taken to measure testosterone and estrogen levels. Furthermore, the testis was surgically removed and macroscopically, histologically, and immunohistochemically examined which lead to the diagnosis of testicular yolk sac tumor. The second testis was descended until the age of nine months and histology revealed impaired spermatogenesis. This report provides the first sonographic images of bovine testicular yolk sac tumor as well as the first information about hormone levels in calves with this neoplasm. It also shows the importance to combine anamnesis, histomorphological, and immunohistochemical findings in order to diagnose yolk sac tumors when pathognomonic structures are not present.

Combined effects of androgen anabolic steroids and physical activity on the hypothalamic-pituitary-gonadal axis.

Analysing effects of pharmaceutical substances and training on feedback mechanisms of the hypothalamic-pituitary-gonadal axis may be helpful to quantify the benefit of strategies preventing loss of muscle mass, and in the fight against doping. In this study we analysed combined effects of anabolic steroids and training on the hypothalamic-pituitary-gonadal axis. Therefore intact male Wistar rats were dose-dependently treated with metandienone, estradienedione and the selective androgen receptor modulator (SARM) S-1. In serum cortisol, testosterone, 17ß-estradiol (E2), prolactin, inhibin B, follicle-stimulating hormone (FSH), luteinizing hormone (LH), Insulin-like growth factor 1 (IGF-1), and thyroxine (T4) concentrations were determined. Six human volunteers were single treated with 1-androstenedione. In addition abusing and clean body builders were analysed. Serum concentrations of inhibin B, IGF-1, cortisol, prolactin, T4, thyroid-stimulating hormone (TSH), testosterone and LH were determined. In rats, administration of metandienone, estradienedione and S-1 resulted in an increase of muscle fiber diameter. Metandienone and estradienedione but not S-1 administration significantly decreases LH and inhibin B serum concentration. Administration of estradienedione resulted in an increase of E2 and S-1 in an increase of cortisol. Single administration of 1-androstenedione in humans decreased cortisol and inhibin B serum concentrations. LH was not affected. In abusing body builders a significantly decrease of LH, TSH and inhibin B and an increase of prolactin, IGF-1 and T4 was detected. In clean body builders only T4 and TSH were affected.

Lifelong exposure to dietary isoflavones reduces risk of obesity in ovariectomized Wistar rats.

SCOPE: Traditional Asian diet rich in soy isoflavones (ISOs) is discussed to be linked to a lower obesity prevalence. In lifelong and short-term exposure scenarios we investigated effects of an ISO-rich diet on the body composition and development of obesity in female rats. METHODS AND RESULTS: Female Wistar rats grew up on ISO-free or ISO-rich control diet (CON ISO: 467 mg/kg diet). Starting postnatal day 83, ovariectomized and intact animals received high calorie Western diet (WD) in the absence or presence of ISO (WD ISO: 431 mg/kg diet) for 12 weeks to induce obesity or maintained on respective control diet (CON). One group starting ISO exposure after ovariectomy mimics short-term ISO exposure in postmenopausal Western women. Lifelong but not short-term ISO exposure resulted in reduced body weight, visceral fat mass, serum leptin, and smaller adipocytes. ISO decreased hepatic SREBP-1c, ACC, FAS, and PPARγ mRNA expression in nonobese animals. Moreover, ovariectomy reduced skeletal muscle weight, which was antagonized by both short-term and lifelong ISO exposure. CONCLUSION: Our results indicate that in female rats lifelong but not short-term ISO intake reduces the risk to develop obesity. Furthermore, lifelong and short-term ISO exposure may antagonize loss of skeletal muscle mass induced by ovariectomy.

Elevated testosterone levels in a racing horse due to an XY testicular disorder of sexual development.

A female thoroughbred successful in horse racing was positively tested for high testosterone values. This horse neither showed stallion-like-behaviour nor signs of ambiguous external genitalia. The karyotype of this horse was 2n = 64,XY and the sex-determining region of Y (SRY) PCR was positive. Hair samples tested for naturally testosterone revealed values normal for stallions, and tests for eight synthetic testosterone esters remained negative. The phenotype, ultrasonographic examination, hormone status, cytogenetic evaluation and molecular diagnostics lead to the diagnosis of an XY testicular disorder of sexual development (DSD) due to a complete androgen insensitivity syndrome. To our knowledge this is the first report about a thoroughbred in racing sports with an XY testicular disorder of sexual development. To date, intersex racing horses have never been described in thoroughbreds or a regulation for intersexes in regard to horse races has been issued.

Effects of Inhibiting Dipeptidyl Peptidase-4 (DPP4) in Cows with Subclinical Ketosis.

The inhibition of dipeptidyl peptidase-4 (DPP4) via specific inhibitors is known to result in improved glucose tolerance and insulin sensitivity and decreased accumulation of hepatic fat in type II diabetic human patients. The metabolic situation of dairy cows can easily be compared to the status of human diabetes and non-alcoholic fatty liver. For both, insulin sensitivity is reduced, while hepatic fat accumulation increases, characterized by high levels of non-esterified fatty acids (NEFA) and ketone bodies.Therefore, in the present study, a DPP4 inhibitor was employed (BI 14332) for the first time in cows. In a first investigation BI 14332 treatment (intravenous injection at dosages of up to 3 mg/kg body weight) was well tolerated in healthy lactating pluriparous cows (n = 6) with a significant inhibition of DPP4 in plasma and liver. Further testing included primi- and pluriparous lactating cows suffering from subclinical ketosis (ß-hydroxybutyrate concentrations in serum > 1.2 mM; n = 12). The intension was to offer effects of DPP4 inhibition during comprehensive lipomobilisation and hepatosteatosis. The cows of subclinical ketosis were evenly allocated to either the treatment group (daily injections, 0.3 mg BI 14332/kg body weight, 7 days) or the control group. Under condition of subclinical ketosis, the impact of DPP4 inhibition via BI 14332 was less, as in particular ß-hydroxybutyrate and the hepatic lipid content remained unaffected, but NEFA and triglyceride concentrations were decreased after treatment. Owing to lower NEFA, the revised quantitative insulin sensitivity check index (surrogate marker for insulin sensitivity) increased. Therefore, a positive influence on energy metabolism might be quite possible. Minor impacts on immune-modulating variables were limited to the lymphocyte CD4+/CD8+ ratio for which a trend to decreased values in treated versus control animals was noted. In sum, the DPP4 inhibition in cows did not affect glycaemic control like it is shown in humans, but was able to impact hyperlipemia, as NEFA and TG decreased.

Influence of transrectal and transabdominal ultrasound examination on salivary cortisol, heart rate, and heart rate variability in mares.

Pregnancy diagnostics in equine reproduction are routinely performed using transrectal ultrasonography, although it is also possible to visualize the fetus by transabdominal ultrasound examinations from the 90th day of gestation onward. We hypothesized that ultrasound examinations may stress the mare and that the gestational stage status and lactation may influence the mare's stress reaction. To investigate the stress reaction, 25 thoroughbred mares of different age, pregnancy and lactational status underwent a transrectal examination. In pregnant mares, an additional transabdominal examination was performed. Salivary cortisol concentration, mean heart rate, and heart rate variability of mares were assessed to evaluate the reactions of hypothalamic-pituitary-adrenal (HPA) axis and of the autonomic nervous system. Significant differences were observed between lactating and nonlactating mares; with a lower responsiveness to stress in lactating mares. The transrectal ultrasound examination in nonlactating mares induced a significant increase in salivary cortisol (P < 0.05), and in the heart rate variability parameter, ratio of low to high frequencies (P < 0.05). This reflects an activation of the HPA axis and a shift to more sympathetic dominance. In contrast, a transabdominally performed pregnancy check did not induce an activation of the HPA axis over basal level but increased the mean heart rate and low to high frequency ratio. The results of this study indicate that checks of advanced pregnancies can be easily performed by transabdominal ultrasonography. With regard to animal welfare, this technique should be preferred during midgestation in nonlactating mares.

Antepartal insulin-like growth factor concentrations indicating differences in the metabolic adaptive capacity of dairy cows.

Cows with different Insulin-like Growth Factor-I (IGF-I) concentrations showed comparable expression levels of hepatic growth hormone receptor (GHR). Suppressor of cytokine signaling 2 (SOCS2), could be responsible for additional inhibition of the GHR signal cascade. The aims were to monitor cows with high or low antepartal IGF-I concentrations (IGF-I(high) or IGF-I(low)), evaluate the interrelationships of endocrine endpoints, and measure hepatic SOCS2 expression. Dairy cows (n = 20) were selected (240 to 254 days after artificial insemination (AI)). Blood samples were drawn daily (day -17 until calving) and IGF-I, GH, insulin, thyroid hormones, estradiol, and progesterone concentrations were measured. Liver biopsies were taken (day 264 ± 1 after AI and postpartum) to measure mRNA expression (IGF-I, IGFBP-2, IGFBP-3, IGFBP-4, acid labile subunit (ALS), SOCS2, deiodinase1, GHR1A). IGF-I concentrations in the two groups were different (p < 0.0001). However, GH concentrations and GHR1A mRNA expression were comparable (p > 0.05). Thyroxine levels and ALS expression were higher in the IGF-I(high) cows compared to IGF-I(low) cows. Estradiol concentration tended to be greater in the IGF-I(low) group (p = 0.06). It was hypothesized that low IGF-I levels are associated with enhanced SOCS2 expression although this could not be decisively confirmed by the present study.