[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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

Estrogen receptor beta is involved in skeletal muscle hypertrophy induced by the phytoecdysteroid ecdysterone.

SCOPE: The phytoectysteroid ecdysterone (Ecdy) was reported to stimulate protein synthesis and enhance physical performance. The aim of this study was to investigate underlying molecular mechanisms particularly the role of ER beta (ERß). RESULTS: In male rats, Ecdy treatment increased muscle fiber size, serum IGF-1 increased, and corticosteron and 17ß-estradiol (E2) decreased. In differentiated C2C12 myoblastoma cells, treatment with Ecdy, dihydrotestosterone, IGF-1 but also E2 results in hypertrophy. Hypertrophy induced by E2 and Ecdy could be antagonized with an antiestrogen but not by an antiandrogen. In HEK293 cells transfected with ER alpha (ERα) or ERß, Ecdy treatment transactivated a reporter gene. To elucidate the role of ERß in Ecdy-mediated muscle hypertrophy, C2C12 myotubes were treated with ERα (ALPHA) and ERß (BETA) selective ligands. Ecdy and BETA treatment but not ALPHA induced hypertrophy. The effect of Ecdy, E2, and BETA could be antagonized by an ERß-selective antagonist (ANTIBETA). In summary, our results indicate that ERß is involved in the mediation of the anabolic activity of the Ecdy. CONCLUSION: These findings provide new therapeutic perspectives for the treatment of muscle injuries, sarcopenia, and cachectic disease, but also imply that such a substance could be abused for doping purposes.

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.

Peripheral blood leukocytes of cows with subclinical endometritis show an altered cellular composition and gene expression.

Subclinical endometritis (SCE) is an important postpartum disease in dairy cows, but conventional cytobrush diagnosis often gives imprecise results. The aim of this study was to analyze disease-associated changes in peripheral blood as potential diagnostic parameters. Cellular subpopulations of blood leukocytes from cows with or without SCE (45-55 days postpartum) were flow-cytometrically quantified. Gene expression of whole blood leukocytes was assessed by PAXgene analysis. Subclinical endometritis cows showed significantly higher number of blood mononuclear cells and neutrophils. Among mononuclear cells, numbers of B-cells, NK-cells, and CD172a-positive monocytes were significantly elevated. Compared with non-SCE cows, blood leukocytes of SCE cows significantly expressed higher copy numbers of CXCL8, TNF, and IL12. To test whether circulating plasma factors are responsible for these changes, leukocytes, polymorphonuclear cells, and monocyte subpopulations (classical, intermediate, nonclassical) of healthy cows were stimulated with plasma of SCE and non-SCE cows. Although gene expression of whole leukocytes and polymorphonuclear cells remained unaltered, plasma from SCE animals significantly elevated expressed messenger RNA copy numbers of CXCL8, CXCL1, and IL1B in intermediate monocytes. In conclusion, elevated number of selected mononuclear subpopulations in peripheral blood and enhanced expression of distinct genes encoding for inflammatory mediators in blood leukocytes reflect the subclinical uterine inflammatory process in cows. Whether the observed changes in the periphery of SCE cows are the consequence of the uterine inflammatory process, or whether they affect the pathogenesis of the disease is currently unknown.

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.

Lifelong obesity in a polygenic mouse model prevents age- and diet-induced glucose intolerance- obesity is no road to late-onset diabetes in mice.

AIMS/HYPOTHESIS: Visceral obesity holds a central position in the concept of the metabolic syndrome characterized by glucose intolerance in humans. However, until now it is unclear if obesity by itself is responsible for the development of glucose intolerance. METHODS: We have used a novel polygenic mouse model characterized by genetically fixed obesity (DU6) and addressed age- and high fat diet-dependent glucose tolerance. RESULTS: Phenotype selection over 146 generations increased body weight by about 2.7-fold in male 12-week DU6 mice (P<0.0001) if compared to unselected controls (Fzt:DU). Absolute epididymal fat mass was particularly responsive to weight selection and increased by more than 5-fold (P<0.0001) in male DU6 mice. At an age of 6 weeks DU6 mice consumed about twice as much food if compared to unselected controls (P<0.001). Absolute food consumption was higher at all time points measured in DU6 mice than in Fzt:DU mice. Between 6 and 12 weeks of age, absolute food intake was reduced by 15% in DU6 mice (P<0.001) but not in Fzt:DU mice. In both mouse lines feeding of the high fat diet elevated body mass if compared to the control diet (P<0.05). In contrast to controls, DU6 mice did not display high fat diet-induced increases of epididymal and renal fat. Control mice progressively developed glucose intolerance with advancing age and even more in response to the high fat diet. In contrast, obese DU6 mice did neither develop a glucose intolerant phenotype with progressive age nor when challenged with a high fat diet. CONCLUSIONS/INTERPRETATION: Our results from a polygenic mouse model demonstrate that genetically pre-determined and life-long obesity is no precondition of glucose intolerance later in life.

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.

Effects of meloxicam and flunixin on pain, stress and discomfort in male piglets during and after surgical castration.

Surgical castration of young male piglets is now a generally accepted cause of serious distress and impairment of animal welfare. Awareness of this problem has created the moral commitment to seek for practical and more humane alternatives. As one possible alternative, the application of analgesics has been installed in Germany as an interim solution by the QS system, thus mandatory for the majority of German pig producers.Two analgesics have been authorised for this purpose. Both have been shown a significant positive impact on cortisol levels if administered pre-operatively. However, their effects on pain, stress and discomfort during castration, and on the post-castration period are conflicting. Thus, the aim of the present study was to compare the effects of Meloxicam and Flunixin on cortisol levels, behavioural indices, vocalisation, and wound healing of surgical castrated piglets in the field. There was no difference in vocalisation during castration in analgesic treated and untreated piglets. Piglets castrated under analgesia still had significantly elevated serum cortisol levels 30 min post castration, when compared to the sham castrated group. Both analgesics led to a significant impairment of behavioural indices and wound healing. It is concluded that analgesics can improve the welfare of piglets during the first part of the post-castration period. However, the benefits may be considered small and may not meet the requirements of the EU. Hence it is of high importance to prevent the interim practice of surgical castration of male piglets under analgesics from becoming implemented as a permanent condition in pig production.

Distribution and viability of spermatozoa in the canine female genital tract during post-ovulatory oocyte maturation.

BACKGROUND: Unlike other domestic mammals, in which metaphase-II oocytes are ovulated, canine ovulation is characterized by the release of primary oocytes, which may take 12 to up to 36 hours. Further 60 hours are needed for maturation to secondary oocytes which then remain fertile for about 48 hours. Oestrus takes 7 to 10 days on average and may start as early as a week before ovulation. This together with the prolonged process of post-ovulatory oocyte maturation requires an according longevity of spermatozoa in the female genital tract in order to provide a population of fertile sperm when oocytes have matured to fertilizability. Therefore the distribution and viability of spermatozoa in the bitch genital tract was examined during post-ovulatory oocyte maturation. METHODS: Thirteen beagle bitches were inseminated on the day of sonographically verified ovulation with pooled semen of two beagle dogs containing one billion progressively motile spermatozoa. Ovariohysterectomy was performed two days later (group 1, n = 6) and four days later (group 2, n = 7). The oviduct and uterine horn of one side were flushed separately and the flushing's were checked for the presence of gametes. The oviducts including the utero-tubal junction and the uterine horns, both the flushed and unflushed, were histologically examined for sperm distribution. RESULTS: The total number of spermatozoa recovered by flushing was low and evaluation of viability was limited. Prophase-I oocytes were collected from oviduct flushing in group 1, whereas unfertilized metaphase-II oocytes were detected in group 2. From day 2 to day 4 after ovulation a significant decrease in the percentage of glands containing sperm (P<0.05) and a marked reduction of the mean sperm number in uterine horn glands were observed. A concomitant diminution of spermatozoa was indicated in the utero-tubal junction accompanied by a slight increase in sperm numbers in the mid oviduct. CONCLUSIONS: Oocyte maturation to metaphase-II stage is accompanied by a continuous sperm detachment and elimination in the uterine horns. Entrance of spermatozoa into the caudal oviduct seems to be steadily controlled by the utero-tubal junction thus providing a selected sperm population to be shifted towards the site of fertilization when oocyte maturation is completed.

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.