Effect of IGFBP-4 during In Vitro Maturation on Developmental Competence of Bovine Cumulus Oocyte Complexes.

Insulin-like growth factors (IGFs) are essential for oocyte maturation. Their bioavailability is regulated by their respective binding proteins (IGFBPs) and proteases. IGFBP-4 blocks the biological effects of IGFs. High IGFBP-4 expression has been associated with follicle atresia. We hypothesized that IGFBP-4 affects oocyte developmental competence during maturation. Therefore, the aim of this study was to examine the effect of IGFBP-4 on the developmental rate of bovine cumulus-oocyte complexes (COCs) during in vitro embryo production. Abattoir-derived COCs were matured with rbIGFBP-4 (2000, 540, and 54 ng/mL) compared to a control. Cumulus expansion, oocyte maturation, cleavage, blastocyst, and hatching rates were evaluated. Furthermore, blastocyst gene expression of SOCS2, STAT3, SLC2A1, SLCA3, BAX, and POU5F1 transcripts were quantified using RT-qPCR. No statistical differences were detected among the groups for cumulus expansion, maturation, cleavage, blastocyst rates, or all gene transcripts analyzed. However, at day 8 and 9, the number of total hatching and successfully hatched blastocysts was lower in 2000 ng/mL rbIGFBP-4 compared to the control (day 8: total hatching: 17.1 ± 0.21 vs. 31.2 ± 0.11%, p = 0.02 and hatched blastocyst 6.7 ± 0.31 vs. 21.5 ± 0.14%, p = 0.004; day 9 total hatching 36.4 ± 0.18 vs. 57.7 ± 0.10%, p = 0.009 and hatched blastocyst 18.2 ± 0.21 vs. 38.1 ± 0.11%, p = 0.004). We concluded that high concentrations of rbIGFBP-4 might negatively affect the subsequent ability of the embryo to hatch and possibly compromise further elongation.

Comparison of Uterine Involution and the Resumption of Ovarian Cyclicity between Lame and Sound Holstein Cows.

The aim of the study was to examine the effect of lameness and energy status on the involution of the uterus and the resumption of ovarian cyclicity in dairy cows. Lame (lameness score of four and the presence of hoof lesions, n = 22) and sound (normal gait and the absence of hoof lesions, n = 25) multiparous cows with healthy puerperium were enrolled simultaneously in the study and were monitored from day 10 antepartum (ap) to day 50 post-partum (pp). Ultrasonography of the cervix, the formerly gravid uterine horn and the ovarian structures was performed on d 8, 11, 14, 23, 30, and 42 pp. Blood sampling for progesterone, ß-hydroxybutyrate (BHBA), and non-esterified fatty acids (NEFAs) was used to assess cyclicity and energy status. Lame compared to sound cows had higher NEFA concentrations on day 14 pp (0.54 ± 0.05 vs. 0.37 ± 0.05, respectively, p = 0.005), delayed involution of the cervix and the formerly pregnant uterine horn (p = 0.0003 and p = 0.02, respectively), lower ovulation rates within the experimental period (63.6% vs. 88%, respectively, p = 0.05), and higher rates of atresia or cyst formation on day 50 pp (36.4% vs. 12%, respectively, p = 0.05). Independently of lameness status, cows with high NEFA concentrations had lower ovulation rates within the experimental period (65.5% vs. 94.4%, p = 0.02), lower normal ovarian activity on day 50 pp (58.6% vs. 88.9%, p = 0.03), and higher rates of atresia or cyst formation on day 50 pp (34.5% vs. 5.6%, p = 0.02) compared to cows with optimal NEFA concentrations. Furthermore, an interaction between lameness and increased NEFA concentrations was observed regarding the ovulation rate within the experimental period and the percentage of atresia or cyst formation on day 50 pp. Sound cows with low NEFA levels had the lowest mean cervical diameter compared to cows with lameness (both with elevated and optimal NEFA concentrations, p = 0.009 and p = 0.002, respectively). Conclusively, lameness during puerperium negatively affected ovarian function and uterine involution. These effects were exacerbated (through interaction or cumulation) in relation to elevated NEFA concentrations.

Comparative cryopreservation of bovine and porcine primary hepatocytes.

The isolation of primary hepatocytes from liver tissue of farm animals yields a very high number of cells, and a part of them can be stored by cryopreservation for future experiments. As no experience exists with the cryopreservation of hepatocytes from cattle, our study aimed at the cryopreservation of bovine hepatocytes by use of different protocols compared with the cryopreservation of hepatocytes from pig. We tested different freezing media (William's Medium E vs. University of Wisconsin solution), cryoprotectants (dimethyl sulfoxide with vs. without trehalose as additional additive), freezing systems (standard freezing container vs. controlled-rate freezer) and freezing times (4 vs. 28 d). These tests identified a general influence of species and freezing systems, whereas the influence of freezing media, trehalose additive and freezing time was less or not obvious. In this regard, we determined a mean recovery of 30% of bovine hepatocytes and 55% of porcine hepatocytes cryopreserved in a controlled-rate freezer, whereas the rates were about 10% less when hepatocytes were frozen in a standard freezing container. In accordance with this observation, the cultivation of cryopreserved hepatocytes from cattle was less effective than that of porcine hepatocytes. Hepatocytes from cattle can be successfully cryopreserved and partially cultured after cryopreservation but with lower percentage than porcine hepatocytes.

Comparative Assessment of the Stress Response of Cattle to Common Dairy Management Practices.

While studies have been conducted examining the stress response of dairy cattle to individual acute and chronic stressors, the results are difficult to compare due to differences in study design and analysis methods. The aim of the present study was to conduct a comparative assessment of the impact of eight common stimuli: artificial insemination (AI), embryo transfer (ET), morning milking (MM), evening milking (EM), veterinary examination (VE), ultrasound examination (US), hoof trimming (HT), and natural breeding (NB) on the hypothalamic-pituitary-adrenal axis and milk production of 24 Holstein-Friesian cattle. After random allocation into control and treatment groups, a total of five blood samples were collected 40 min (Ba1) and 20 min (Ba2) prior to stimulus application, immediately following the stimulus (St), as well as 20 min (Re1) and 40 min (Re2) post-stimulus. A comparison between the overall serum cortisol concentrations in the treatment groups showed a significant difference between HT to AI (p = 0.006), ET (p = 0.010), MM (p = 0.021), VE (p = 0.009), EM (p = 0.007), and US (p = 0.010), except for NB (p = 0.542). There is no significant difference between the control groups (p > 0.05). The stimuli HT (p < 0.001) and NB (p < 0.001) showed significant increases in cortisol following stimulus application, and the levels failed to decrease significantly by sample Re2. No significant differences in daily milk yield (kg) were measured amongst the tested stimuli (p = 0.472) nor amongst the groups 'Control', 'Treatment' and 'no stimulus' (p = 0.350). In conclusion, when factors such as increased physical activity, novel social interaction, sexual arousal, and a more intense restriction of movement are present, the animal's perceived controllability and predictability might decrease, affecting the animal's response to stress. Treatments carried out while animals are restrained in a headlock while remaining within their regular group likely result in a less intense activation of the HPA axis.

Dual use of breeding stallions is possible without affecting the sperm quality.

Artificial insemination (AI) is commonly used in the equine industry to enhance the genetic value in breeding programs and to effectively utilize ejaculates. Many stallions are used as breeding stallions as well as in high-level sports competitions to improve their market value. The goal of the present study was to investigate whether this dual use of stallions influences the animals´ stress levels and/or the quality of their ejaculates. For this purpose, 18 stallions were grouped into two categories: breeding stallions with (BSC = breeding stallion competition), and breeding stallions without secondary use in competitions (BS = breeding stallion). Two ejaculates were collected at a one-week interval and analysed with an extended spectrum of spermatological methods. Furthermore, saliva, as well as seminal plasma samples were taken, and the concentration of cortisol therein was determined. Additionally, dehydroepiandrosterone (DHEA) and the cortisol/DHEA ratio were analysed and calculated for seminal plasma. After statistical analysis of the correlations and interdependences between the two groups, the results showed that the BSC group had significantly higher saliva cortisol levels (p = .027) and tendentially higher DHEA concentrations in their seminal plasma (p = .056). No difference between BS and BSC could be found in regard to the sperm quality parameters and the cortisol concentration in seminal plasma samples. It can be concluded that while active participation in competitions represents a stress factor, the dual use of stallions in breeding programs and sports competitions is possible without negative effects on their sperm quality.

An Approach for Investigating Sexual Maturity in Wild Boar Males: Testosterone and 17ß-Estradiol Analysis.

Analyses of sexual steroid hormones in wild boars are rarely described. Testosterone (T) and 17ß-estradiol (E2) concentrations are useful to recognize sexual maturation. As threshold values for this species are unknown, additional parameters are required. A total of 127 blood samples from wild boar males were collected to measure T and E2. Age and weight were recorded. Thirty-one epididymides were sampled too. Males were sorted into pre-and postpubertal groups based on the absence/presence of spermatozoa in epididymides and on morphological data following previous results. Forty-four males were prepubertal: the mean age and weight were 10 months and 23 kg, respectively. They showed no spermatozoa. The mean concentrations of T and E2 were 1.2 ± 1.2 ng/mL and 39.7 ± 120.3 pg/mL, respectively. Sixty-six males were postpubertal, twenty-nine of which presented spermatozoa. Their mean concentration of T was 7.6 ± 6.3 ng/mL and E2 was 664.3 ± 250.4 pg/mL. Seventeen samples could not be defined; the hormone concentrations between the two groups suggested a transitional phase consistent with puberty. Wild boars before 12 months of age had high hormone levels like older boars, indicating that they could attempt to reproduce. Hormones at the end of the mating season (January) were high so that reproduction could occur thereafter, shifting farrowing from spring to summer.

Allele-biased expression of the bovine APOB gene associated with the cholesterol deficiency defect suggests cis-regulatory enhancer effects of the LTR retrotransposon insertion.

The insertion of an endogenous retroviral long terminal repeat (LTR) sequence into the bovine apolipoprotein B (APOB) gene is causal to the inherited genetic defect cholesterol deficiency (CD) observed in neonatal and young calves. Affected calves suffer from developmental abnormalities, symptoms of incurable diarrhoea and often die within weeks to a few months after birth. Neither the detailed effects of the LTR insertion on APOB expression profile nor the specific mode of inheritance nor detailed phenotypic consequences of the mutation are undisputed. In our study, we analysed German Holstein dairy heifers at the peak of hepatic metabolic load and exposed to an additional pathogen challenge for clinical, metabolic and hepatic transcriptome differences between wild type (CDF) and heterozygote carriers of the mutation (CDC). Our data revealed that a divergent allele-biased expression pattern of the APOB gene in heterozygous CDC animals leads to a tenfold higher expression of exons upstream and a decreased expression of exons downstream of the LTR insertion compared to expression levels of CDF animals. This expression pattern could be a result of enhancer activity induced by the LTR insertion, in addition to a previously reported artificial polyadenylation signal. Thus, our data support a regulatory potential of mobile element insertions. With regard to the phenotype generated by the LTR insertion, heterozygote CDC carriers display significantly differential hepatic expression of genes involved in cholesterol biosynthesis and lipid metabolism. Phenotypically, CDC carriers show a significantly affected lipomobilization compared to wild type animals. These results reject a completely recessive mode of inheritance for the CD defect, which should be considered for selection decisions in the affected population. Exemplarily, our results illustrate the regulatory impact of mobile element insertions not only on specific host target gene expression but also on global transcriptome profiles with subsequent biological, functional and phenotypic consequences in a natural in-vivo model of a non-model mammalian organism.

Brief Research Report: Effect of Triiodothyronine on Hepatic Growth Hormone Receptor Expression in Primary Bovine Hepatocytes.

In previous studies, triiodothyronine (T3) was found to be lower in cows with ketosis and an effect of T3 on Growth Hormone Receptor (GHR) expression is described, e. g., in a human hepatoma cell line. Therefore, this study aimed to test whether T3 affects GHR messenger RNA (mRNA) expression in a well-established bovine hepatocyte model. Hepatocytes were kept in a sandwich culture and stimulated for 6 days with constant (10 µg/ml) or decreasing (from 10 to 5 µg/ml) T3 concentrations, and GHR, as well as IGF-1 mRNA expression, was measured using real-time polymerase chain reaction (RT-PCR). We could confirm in vitro that T3 has a stimulatory effect on GHR1A mRNA expression.

Clinical effect of buprenorphine or butorphanol, in combination with detomidine and diazepam, on sedation and postoperative pain after cheek tooth extraction in horses.

The objective of this study was to compare effects of butorphanol (BUT) or buprenorphine (BUP), in combination with detomidine and diazepam, on the sedation quality, surgical conditions, and postoperative pain control after cheek tooth extraction in horses, randomly allocated to 2 treatment groups (BUT: n = 20; BUP: n = 20). A bolus of detomidine (15 µg/kg, IV) was followed by either BUP (7.5 µg/kg, IV) or BUT (0.05 mg/kg, IV). After 20 min, diazepam (0.01 mg/kg, IV) was administered and sedation was maintained with a detomidine IV infusion (20 µg/kg/h), with rate adjusted based on scores to 5 variables. All horses received a nerve block (maxillary or mandibular), and gingival infiltration with mepivacaine. Sedation quality was assessed by the surgeon from 1 (excellent) to 10 (surgery not feasible). A pain scoring system (EQUUS-FAP) was used to assess postoperative pain. Serum cortisol concentrations and locomotor activity (pedometers) were measured. Horses in BUP and BUT required a median detomidine infusion rate of 30.2 µg/kg/h (20 to 74.4 µg/kg/h) and 32.2 µg/kg/h (20 to 48.1 µg/kg/h), respectively (P = 0.22). Horses in the BUP group had better sedation quality (P < 0.05) during surgery and higher step counts (P < 0.001) postoperatively. Buprenorphine combined with detomidine provided a more reliable sedation than butorphanol. However, the EQUUS-FAP pain scale became unreliable because of BUP-induced excitement behavior.

Effet clinique de la buprénorphine ou du butorphanol, en association avec la détomidine et le diazépam, sur la sédation et la douleur postopératoire après extraction de dents jugales chez le cheval. L'objectif de cette étude était de comparer les effets du butorphanol (BUT) ou de la buprénorphine (BUP), en association avec la détomidine et le diazépam, sur la qualité de la sédation, les conditions chirurgicales et la gestion de la douleur postopératoire après extraction des dents jugales chez les chevaux, répartis au hasard dans deux groupes de traitement (BUT : n = 20; BUP : n = 20). Un bolus de détomidine (15 µg/kg, IV) a été suivi soit de BUP (7,5 µg/kg, IV) soit de BUT (0,05 mg/kg, IV). Après 20 min, du diazépam (0,01 mg/kg, IV) a été administré et la sédation a été maintenue avec une perfusion IV de détomidine (20 µg/kg/h), avec un taux ajusté en fonction des scores de cinq variables. Tous les chevaux ont reçu un bloc nerveux (maxillaire ou mandibulaire) et une infiltration gingivale avec de la mépivacaïne. La qualité de la sédation a été évaluée par le chirurgien de 1 (excellent) à 10 (chirurgie impossible). Un système de notation de la douleur (EQUUS-FAP) a été utilisé pour évaluer la douleur postopératoire. Les concentrations sériques de cortisol et l'activité locomotrice (podomètres) ont été mesurées.Les chevaux en BUP et BUT ont nécessité un débit médian de perfusion de détomidine de 30,2 µg/kg/h (20 à 74,4 µg/kg/h) et 32,2 µg/kg/h (20 à 48,1 µg/kg/h), respectivement (P = 0,22). Les chevaux du groupe BUP avaient une meilleure qualité de sédation (P < 0,05) pendant la chirurgie et un nombre de pas plus élevé (P < 0,001) après l'opération. La buprénorphine associée à la détomidine a fourni une sédation plus fiable que le butorphanol. Cependant, l'échelle de douleur EQUUS-FAP est devenue peu fiable en raison du comportement d'excitation induit par le BUP.(Traduit par Dr Serge Messier).

Transrectal three-dimensional fetal volumetry in early pregnant mares: Relationships between maternal factors and equine fetal volume measurements.

The aim of this study was to investigate the effects of maternal, hormonal, and fetal factors on early fetal volume (FV) measurements in mares obtained by three-dimensional (3D) ultrasound. Furthermore, postpartum parameters were explored in regard to their association with early FV. For this purpose, 149 German warmblood mares that were artificially inseminated and confirmed to be pregnant between days 14-16 of gestation, were examined transrectally at day 45 ± 1 of gestation with the portable 3D ultrasound device Voluson® i (GE, Zipf, Austria). FV was calculated by using the extension software Virtual Organ Computer-aided AnaLysis (VOCAL™). Two different mixed linear models were used to analyze associations between the investigated maternal and fetal factors and the FV. Explanatory variables investigated in the first model were: maternal age, parity, maternal weight, and body condition score, type of pregnancy (recipient or biological mother), barren status (lactating or non-lactating), fetal sex, progesterone (P4) and equine chorionic gonadotropin (eCG) concentrations; and in the second model outcome variables such as gestational length, birth weight, placental weight, fetal sex, and abortion were included in the analysis. The final models revealed a significant relation between FV and eCG (b = 0.011, P = 0.030), as well as with P4 (b = -0.053, P = 0.016), but interestingly P4 was negatively related to FV. Fetal sex showed the most prominent effect on FV (b = -0.256, P = 0.039), with female fetuses being smaller than male fetuses. In the second model none of the investigated parameters were related to early FV except for fetal sex (b = -0.328, P = 0.047), again with female fetuses being smaller. In summary, it was found that FV is related with eCG, P4 and fetal sex, but was no suitable predicting factor for the investigated outcome parameters. Furthermore, the findings suggest that sex specific growth differences exist already in early gestation. The detailed biological mechanism by which P4 and eCG affect fetal size has to be investigated in prospective studies.

An approach to assess stress in response to drive hunts using cortisol levels of wild boar (Sus scrofa).

Hunting can easily be linked to stress in wildlife. Drive hunts performed two to three times in one area during the respective hunting period, are thought to decrease the pressure hunting places on wildlife. Nevertheless, the expression of cortisol-one of the main mammalian stress hormones-is considered to have negative impacts on animals' well-being if expressed excessively, which may occur during some (especially repeated) hunting events. We explored the effect of drive hunts on cortisol levels in wild boar in Lower Saxony, Germany, compared these cortisol levels to reference values given by a similar study, and investigated the effect of age, sex, and pregnancy. Blood collected from wild boar shot on drive hunts was analysed using a radioimmunoassay. As expected, we observed elevated cortisol levels in all samples, however, we still found significant differences between age groups and sexes, as well as an influence of pregnancy on cortisol levels. The effect of drive hunts on cortisol levels appears to be weaker than predicted, while the effects of other variables, such as sex, are distinct. Only half of the evaluated samples showed explicitly increased cortisol levels and no significant differences were found between sampling months and locations. Group living animals and pregnant females showed significantly higher cortisol levels. The impact of hunting is measurable but is masked by natural effects such as pregnancy. Thus, we need more information on stress levels in game species.

An investigation on the relevance of prolactin, insulin-like growth factor-1 and 25-hydroxyvitamin D3 (25-OHD3 ) in canine benign prostatic hyperplasia in a predisposed breed model.

Serum concentrations of prolactin (PRL), insulin-like growth factor-1 (IGF-1) and 25 hydroxyvitamin D3 (25-OHD3 ) were analysed to investigate their possible involvement in the pathogenesis of benign prostatic hyperplasia (BPH). For this, dogs of the Rhodesian Ridgeback (RR) breed were used because of a verified breed disposition for the development of BPH. Labrador Retrievers (LR) served as controls. The prostate gland status was characterised by the prostate gland volume, clinical signs of BPH (haemospermia and sonographic findings) and the plasma concentration of canine prostate-specific arginine esterase (CPSE). Breed specificity in the RR was expressed by a correlation of PRL with breed (p < 0.05). Similar relationships existed in the dogs with normal CPSE (CPSEn) with respect to the IGF-1 concentrations (LR: p < 0.05). The latter were negatively correlated with prostatic volume and age (both p < 0.05). Concentrations of 25-OHD3 were tendentially (p = 0.18) lower in the RR with increased CPSE (CPSEi) compared with the CPSEn LR and RR showing clinical signs of BPH. A negative correlation between serum 25-OHD3 and age (p < 0.05) existed in the CPSEi RR. Proof of 25-OHD3 in prostatic secretion proved to be a breed specific feature in the RR (p < 0.0001). For all RR dogs showing clinical signs of BPH, a close to significant (p = 0.06) positive correlation with prostate gland volume was found. The results of the present study reveal no clear hints towards the significance of PRL and IGF-1 in the pathogenesis of canine BPH. In the RR breed there were indications of a causal relationship with age-dependent changes in the vitamin D metabolism. The data suggest the possibility of preventing or treating canine BPH by administering vitamin D or substances involved in the intraprostatic vitamin D metabolism.

Is Harbor Porpoise (Phocoena phocoena) Exhaled Breath Sampling Suitable for Hormonal Assessments?

Over the last decades, exhaled breath sampling has been established for laboratory analysis in various cetacean species. Due to their small size, the usability of respiratory vapor for hormone assessments was questionable in harbor porpoises (Phocoena phocoena). This pilot study compared three different blow collection devices for their suitability in the field and during laboratory processing: a sterile petri dish covered by a Nitex membrane, as well as sterile 50 mL centrifuge tubes with or without manganese(II) chloride as a stabilizer. Collected exhales varied between three, five or ten, depending on feasibility. Hormones were extracted through an ether mix, followed by centrifugal evaporation and cortisol analysis using an immunoassay. Although close to the lower end of the assay's dynamic range, the ELISA produced results (n = 110, 0.102-0.937 ng/mL). Hence, a simple 50 mL centrifuge tube was determined as the best suited blow collection device, while three consecutive exhales proved sufficient to yield results. These findings are promising regarding the suitability of exhaled breath as a matrix for future endocrine and immune system-related studies in harbor porpoises. If further advanced, blow sampling can become an important, non-invasive tool for studying and monitoring health, stress levels and diseases in harbor porpoises.

In vitro effects of Type I interferons (IFNτ and IFNα) on bovine hepatocytes cultured with or without Kupffer cells.

In cattle, maternal recognition of early pregnancy depends on the effects of the embryonic signal interferon (IFN)-τ. IFN-stimulated genes have been upregulated in the maternal liver during early pregnancy. In this study, primary hepatocyte cell culture models were evaluated for their suitability to test Type I IFN effects invitro. The expression of target genes (interferon-stimulated gene 15 (ISG-15), interferon-induced GTP-binding protein (MX-1), C-X-C motif chemokine 10 (CXCL-10), CXCL-5, insulin-like growth factor 1 (IGF-1), IGF binding protein 2 (IGFBP-2)) was measured using reverse transcription-quantitative polymerase chain reaction in hepatocytes from monoculture or in indirect coculture with Kupffer cells (HKCid) on Days 1, 2, 3 and 4 of culture (n=21 donor cows). Gene expression was also measured on Day 4 after challenging the cultures with recombinant IFNτ, IFNα, progesterone (P4), IFNτ+IFNα or IFNτ+P4 for 6h. A significant increase in the mRNA expression of target genes in hepatocytes was shown in response to stimulation with IFNτ. The Kupffer cells in coculture did not influence the effects of IFNτ in hepatocytes. In conclusion, primary bovine hepatocyte cultures are suitable for stimulation experiments with Type I IFNs and as an extrauterine model for embryo-maternal communication. The proposed endocrine action of IFNτ in the liver may affect maternal metabolism and immune function in the liver.

Hepatic lipidosis: Liver characteristics and acute phase proteins in affected turkeys.

The hepatic lipidosis (HL) in fattening turkeys is a disease has been known for a long time, but the cause and pathogenesis is still not clarified. A recent study reported unexplained high levels of iron in liver tissue of fattening turkeys suffering from HL. In this study, the iron status, possible infectious or inflammatory influences in form of an acute phase reaction and the analysis of fatty acid pattern in liver tissue of turkeys affected by HL were examined. Three cases of HL on three different fattening turkey farms were investigated during the outbreak of the disease. Clinically affected and non-affected animals were subjected to a pathological examination, where the diagnosis HL or non-affected was made. In total, 70 birds were examined (40 with HL, 30 without HL) and blood and liver samples were taken. Additionally, samples from 15 slaughtered birds were taken as a further control group. In liver tissue, the iron content and the content of long-chain fatty acids were determined; in blood samples, ferritin and transferrin were measured. The iron content in liver tissue was more than three times higher for animals with HL than among non-affected animals and the control group. The transferrin levels were lowest for animals with HL, highest in the control group and in between for non-affected animals. The fatty acid pattern in liver tissue of affected animals indicated a shift from polyunsaturated fatty acids to saturated fatty acids and monounsaturated fatty acids compared to the control group and the non-affected animals. Overall, the non-affected animals of a flock affected by HL were similar to the healthy animals of the abattoir. The low acute phase protein levels for animals with HL together with high iron contents could indicate a previous malnutrition/starvation period and/or severe liver damage for those animals suffering from HL.

In vitro effects of lipopolysaccharides on bovine uterine contractility.

Metritis is an important disorder in dairy cows during the early postpartum period. Myometrial contractility is a prerequisite for uterine involution; however, very scanty literature is available about the effect of metritis on this process and endocrine responsiveness. This study was aimed to evaluate the effect of inflammation on uterine contractility in vitro, and the inflammation was induced by incubating myometrial strips with lipopolysaccharides (LPS). Myometrial samples were collected from 17 healthy Holstein Friesian cows during caesarean section. Eight longitudinal strips from each cow were incubated in organ baths with LPS concentrations of 0 (LPS0 ), 0.1 (LPS0.1 ), 1 (LPS1 ) and 10 µg/ml (LPS10 ). Spontaneous contractility and contractility induced by increasing concentrations of oxytocin (10-10 - 10-7  mol/L) were recorded during nine 30-min intervals (T1 to T9). The minimum amplitude (minA), maximum amplitude (maxA), mean amplitude (meanA) and area under the curve (AUC) were calculated for each time interval. LPS had an effect (p ≤ .05) on maxA, meanA and AUC. In T1, myometrial strips incubated with LPS0.1 and LPS1 had higher (p ≤ .05) maxA, meanA and AUC than the strips incubated with LPS0 . In T9 without oxytocin, LPS0 led to higher (p ≤ .05) maxA, meanA and AUC than LPS0.1 and LPS1 . In T8 and T9 with oxytocin, LPS1 had lower (p ≤ .05) maxA, meanA and AUC than the other LPS concentrations. Interestingly, the results show that LPS has a transient positive effect on myometrial contractility in vitro and that this effect is dependent on LPS concentration and duration of incubation.

Hepatic Transcriptome Analysis Identifies Divergent Pathogen-Specific Targeting-Strategies to Modulate the Innate Immune System in Response to Intramammary Infection.

Mastitis is one of the major risks for public health and animal welfare in the dairy industry. Two of the most important pathogens to cause mastitis in dairy cattle are Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli). While S. aureus generally induces a chronic and subclinical mastitis, E. coli is an important etiological pathogen resulting in an acute and clinical mastitis. The liver plays a central role in both, the metabolic and inflammatory physiology of the dairy cow, which is particularly challenged in the early lactation due to high metabolic and immunological demands. In the current study, we challenged the mammary glands of Holstein cows with S. aureus or E. coli, respectively, mimicking an early lactation infection. We compared the animals' liver transcriptomes with those of untreated controls to investigate the hepatic response of the individuals. Both, S. aureus and E. coli elicited systemic effects on the host after intramammary challenge and seemed to use pathogen-specific targeting strategies to bypass the innate immune system. The most striking result of our study is that we demonstrate for the first time that S. aureus intramammary challenge causes an immune response beyond the original local site of the mastitis. We found that in the peripheral liver tissue defined biological pathways are switched on in a coordinated manner to balance the immune response in the entire organism. TGFB1 signaling plays a crucial role in this context. Important pathways involving actin and integrin, key components of the cytoskeleton, were downregulated in the liver of S. aureus infected cows. In the hepatic transcriptome of E. coli infected cows, important components of the complement system were significantly lower expressed compared to the control cows. Notably, while S. aureus inhibits the cell signaling by Rho GTPases in the liver, E. coli switches the complement system off. Also, metabolic hepatic pathways (e.g., lipid metabolism) are affected after mammary gland challenge, demonstrating that the liver restricts metabolic tasks in favor of the predominant immune response after infection. Our results provide new insights for the infection-induced modifications of the dairy cow's hepatic transcriptome following mastitis.

Histological Comparison of Testicular Needle Biopsy and En Bloc Samples in Abattoir Calves.

the aim of this study was to test whether a single testicular needle biopsy could provide histological results comparable to en bloc resection histology and whether one biopsy was sufficient to reflect the histology of an entire pair of testicles. Two methods of sample collection were tested on 32 bull calves aged five to eight months to compare histological parameters of needle biopsy with those of en bloc resection samples. One testicular needle biopsy of the right and three en bloc samples of both testicles were collected and compared for the number of tubular cross sections, tubules with elongated spermatids (ES), outer/inner diameter of tubules, thickness of tubular wall, and number of Sertoli cells (SC). Additionally, animal data were considered. No significant differences were found between the left and right testis or among the individual locations of en bloc samples. However, histologically significant differences (Bonferroni-adjusted significance level: p < 0.05/6 = 0.0083) were found between the needle biopsy and en bloc resection regarding the tubular cross sections per visual field (p < 0.05), the outer (p = 0.01) and inner diameter and the thickness of the tubular wall (both p < 0.01). In the SOX9 immunohistochemical staining, no significant differences (p > 0.05) could be observed for SC numbers between needle biopsy and en bloc samples. In conclusion, results of testicular needle biopsy do not have the same validity as the en bloc resection histology. Furthermore, one biopsy is insufficient to reflect the histology of the entire testicular pair.

Correction to: Lack of adverse effects in subchronic and chronic toxicity/carcinogenicity studies on the glyphosate-resistant genetically modified maize NK603 in Wistar Han RCC rats.

In the original publication, the starting point in time for the three feeding trials.

Influence of dental restoration under general anaesthesia on the low-dose dexamethasone suppression test in dogs.

It was the aim of the study to assess the impact of a minor surgical intervention under general anaesthesia on results of a low-dose dexamethasone suppression test (LDDST) in dogs. Five clinically healthy dogs underwent a LDDST (standard protocol) prior and 1, 4, 7, 14 and 28 days after a dental restoration under general anaesthesia. All LDDSTs revealed negative results. On all test days after intervention some dogs had basal cortisol concentrations below the reference range. Accordingly, plasma cortisol concentrations 4 and 8 h after dexamethasone injection were noticeably lower than before surgery and often even below the lower detection limit of 2.0 ng/ml. The study results may indicate a suppressive effect of a minor surgery under general anaesthesia on cortisol measurements during LDDSTs. It may be speculated that this could possibly lead to false negative test results in the postsurgical period, although transfer of these results to clinical cases is subject to limitations.