Evaluation of hormones as candidate early biomarkers of Holstein bull performance around the first semen collection at artificial insemination centres.

Artificial insemination (AI) centres select bulls as calves according to their genetic breeding values and raise them until the first semen collection; yet, a high dropout rate of reared bulls is a problem for AI centres. Potential hormonal indicators of bull sexual maturation (cortisol, dehydroepiandrosterone (DHEA), testosterone, oestradiol, insulin-like growth factor 1 (IGF-1)) were observed and evaluated in relation to the performance parameters to perhaps identify candidate biomarkers allowing an early selection of bulls as suitable sires. Blood samples from 102 German Holstein calves at 4 ± 1, 8 ± 1 and 12 ± 2 months of age from six AI centres were analysed using validated immunoassays for cortisol, DHEA, testosterone, oestradiol and IGF-1. Semen analyses included native and thawed diluted semen. Bulls were classified at the first semen collection into groups with good versus poor performance (GP vs. LP). After 2 years, the subsequent differentiation was done in high (HPP), medium (MPP) and low performance persistency (LPP). Age at first semen collection was an important factor for sperm quality. Cortisol concentrations decreased with age, but the cortisol/DHEA ratio decreased with age only in GP bulls (p < .05). Oestradiol and testosterone concentrations both correlated with libido behaviour (p < .05). Testosterone and IGF-1 concentrations were higher at the time of first semen collection in GP bulls and increased with age (p < .05). In conclusion, testosterone and IGF-1 concentrations at first semen collection are associated with performance at first semen collection and future performance persistency, and might be useful early biomarkers for consistent sperm producing bulls on AI centres.

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.

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).

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.

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 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.

Modulation of growth hormone receptor-insulin-like growth factor 1 axis by dietary protein in young ruminants.

A reduced protein intake causes a decrease in insulin-like growth factor 1 (IGF1) concentrations and modulates Ca homoeostasis in young goats. IGF1 is synthesised by the liver in response to stimulation by growth hormone (GH). Due to rumino-hepatic circulation of urea, ruminants are suitable for investigating the effects of protein reduction despite sufficient energy intake. The present study aimed to investigate the impact of a protein-reduced diet on the expression of components of the somatotropic axis. Male young goats were divided into two feeding groups receiving either a control diet (20 % crude protein (CP)) or a reduced-protein diet (9 % CP). Blood concentrations of IGF1 and GH were measured, and a 24-h GH secretion profile was compiled. Moreover, ionised Ca and insulin concentrations as well as mRNA and protein expression levels of hepatic proteins involved in GH signalling were quantified. Due to the protein-reduced diet, concentrations of ionised Ca, insulin and IGF1 decreased significantly, whereas GH concentrations remained unchanged. Expression levels of the hepatic GH receptor (GHR) decreased during protein reduction. GHR expression was down-regulated due to diminished insulin concentrations as both parameters were positively correlated. Insulin itself might be reduced due to reduced blood Ca levels that are involved in insulin release. The protein-reduced diet had an impact on the expression of components of the somatotropic axis as a disruption of the GH-IGF1 axis brought about by diminished GHR expression was shown in response to a protein-reduced diet.

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.

Rhodesian Ridgebacks have an increased risk to develop benign prostatic hyperplasia.

Benign prostatic hyperplasia (BPH) is an age-dependent primarily non-inflammatory enlargement of the accessory gland in the intact dog. The aim of the present study was to control a previously raised suspicion of a breed-related higher incidence of BPH in dogs of the Rhodesian Ridgeback breed. For this, 18 Labrador Retrievers/LR and 20 Rhodesian Ridgebacks/RR were assigned to the age groups 18-24 months (n = 12), 25-48 months (n = 13) and 49-72 months (n = 13). Prostate gland status was determined by rectal palpation, B-mode ultrasound, calculation of the prostate gland volume and semen analysis regarding haemospermia and was classified according to blood plasma concentrations of canine prostate-specific arginine esterase (CPSE) (normal ≤ 60 ng/ml, increased ≥ 61 ng/ml; Pinheiro et al., 2017). Concentrations of testosterone, 5α-dihydrotestosterone and estradiol were analysed in peripheral blood serum or plasma for detecting breed-specific conditions regarding the endocrine metabolism. Prostatic volume was significantly larger in RR irrespective of the CPSE status. In RR, BPH occurred more frequently and started at an earlier age compared with the LR. Breed-related specificities in steroid metabolism in the RR were indicated by correlations of 5α-dihydrotestosterone and estradiol with age and of testosterone with prostate gland volume. Although the incidence of sonographic signs of BPH and haemospermia did not fit with normal and increased CPSE concentrations, a breed-specific higher incidence of BPH in the RR breed could be clearly verified.

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 2012, a controversial study on the long-term toxicity of a Roundup herbicide and the glyphosate-tolerant genetically modified (GM) maize NK603 was published. The EC-funded G-TwYST research consortium tested the potential subchronic and chronic toxicity as well as the carcinogenicity of the glyphosate-resistant genetically modified maize NK603 by performing two 90-day feeding trials, one with GM maize inclusion rates of 11 and 33% and one with inclusion rates of up to 50%, as well as a 2-year feeding trial with inclusion rates of 11 and 33% in male and female Wistar Han RCC rats by taking into account OECD Guidelines for the testing of chemicals and EFSA recommendations on the safety testing of whole-food/feed in laboratory animals. In all three trials, the NK603 maize, untreated and treated once with Roundup during its cultivation, and the conventional counterpart were tested. Differences between each test group and the control group were evaluated. Equivalence was assessed by comparing the observed difference to differences between non-GM reference groups in previous studies. In case of significant differences, whether the effects were dose-related and/or accompanied by changes in related parameters including histopathological findings was evaluated. It is concluded that no adverse effects related to the feeding of the NK603 maize cultivated with or without Roundup for up to 2 years were observed. Based on the outcome of the subchronic and combined chronic toxicity/carcinogenicity studies, recommendations on the scientific justification and added value of long-term feeding trials in the GM plant risk assessment process are presented.

Bovine embryo elongation is altered due to maternal fatty acid supplementation.

The pre-implantation period is prone to embryonic losses in bovine. Embryo-maternal communication is crucial to support embryo development. Thereby, factors of the uterine fluid (UF) are of specific importance. The maternal diet can affect the UF composition. Since omega 3 fatty acids (omega 3 FA) are considered to be beneficial for reproduction, we investigated if dietary omega 3 FA affected factors in the UF related to embryo elongation. Angus heifers (n = 37) were supplemented with either 450 g of rumen-protected fish oil (omega 3 FA) or sunflower oil (omega 6 FA) for a period of 8 weeks. Following cycle synchronization and artificial insemination, the uteri were flushed post mortem to recover the embryos on day 15 of pregnancy. The UF and tissue samples of endometrium and corpus luteum (CL) were collected. Strikingly, the embryo elongation in the omega 3 group was enhanced compared to the omega 6 group. No differences were observed in uterine prostaglandins, even though the endometrial concentration of their precursor arachidonic acid was reduced in omega 3 compared to omega 6 heifers. The dietary FA neither led to differential expression of target genes in endometrium nor CL nor to a differential abundance of low-density lipoprotein cholesterol, cortisol or amino acids in the UF. Interestingly, the omega 3 group displayed a higher plasma progesterone concentration during luteal growth than the omega 6 group, possibly promoting embryo elongation. Further research should include an ovarian perspective to understand the functional link between dietary omega 3 FA and reproductive outcome.

Nutrient digestibility, organ morphometry and performance in vaccinated or non-vaccinated Lawsonia intracellularis infected piglets.

BACKGROUND: Lawsonia intracellularis is one of the world's most important infectious diseases in pork production with regard to economic losses. So far, studies are missing that describe the effects of a natural infection of piglets on the digestibility of nutrients, possible effects on performance and the morphometrics of the intestine depending on whether piglets are vaccinated, clinically healthy or clinically affected with regard to Lawsonia intracellularis induced diarrhoea. RESULTS: Digestibility studies were performed on a total of 27 eight-week-old piglets with naturally occurring Lawsonia intracellularis infection in a trial with three repetitions. Nine out of 27 animals were vaccinated as suckling pigs with a commercial Lawsonia intracellularis vaccine (vac; Enterisol®Ileitis). Half of the remaining 18 animals were without clinical signs of infection (non-vac/cs-), half showed moderate clinical signs of Lawsonia intracellularis induced diarrhoea (non-vac/cs+). All three groups were fed one identical complete diet ad libitum. Faecal shedding of Lawsonia intracellularis was found in all groups (25 out of 27 animals). Numerically, the mean excretion in the group non-vac/cs + (7.69 ± 1.65 log10 copies/ g faeces) was higher in comparison to the group non-vac/cs- (5.83 ± 2.35 log10 copies/ g faeces) and vaccinated animals (vac: 6.00 ± 2.89log10 copies/ g faeces). The average daily weight gain (ADG; Ø 8.66 day period) differed significantly (vac: 894a ± 73.3, non-vac/cs-: 857ab ± 86.3, non-vac/cs+: 785b ± 137 g/day). The apparent total tract digestibility (ATTD) of nitrogen was significantly lower in clinically affected animals (vac: 83.0a ± 1.72, non-vac/cs-: 83.9a ± 2.03, non-vac/cs+: 80.7b ± 2.57).The total length of the small intestine in clinically affected animals increased significantly (vac: 15.9ab ± 1.57, non-vac/cs-: 14.6b ± 1.12, non-vac/cs+: 16.2a ± 1.37 m). The relative body weight depending on the length of the small intestine was lower for clinically affected animals (vac: 1.72a ± 0.21, non-vac/cs-: 1.83a ± 0.17, non-vac/cs+: 1.56b ± 0.12 kg/m). CONCLUSION: These studies show that clinically moderate L. intracellularis infections lead to significantly lower ADGs in comparison to vaccinated animals. The disease is also found in altered intestinal morphometry and reduced total N digestibility if clinical signs occur.

Sea Buckthorn Pomace Supplementation in the Diet of Growing Pigs-Effects on Fatty Acid Metabolism, HPA Activity and Immune Status.

There is evidence that sea buckthorn, as a source of n-3 polyunsaturated fatty acids (n-3 PUFA), possesses health-enhancing properties and may modulate neuroendocrine and immune functions. In the present study, we investigated the effect of sea buckthorn pomace (SBP) supplementation in the diet of growing German Landrace pigs on fatty acids in the blood and hypothalamus, peripheral immune parameters and mRNA expression of corticotropin-releasing hormone (CRH), mineralocorticoid receptor (MR) and glucocorticoid receptor (GR) in the hypothalamus and spleen. Pigs were fed diets supplemented with 12% of dried SBP or 0% SBP (control group) over an intervention period of eight weeks. The fatty acid profiles in blood plasma were significantly affected by SBP supplementation only for C18:2n-6 and n-6/n-3 PUFA ratio compared with the control group. SBP supplementation did not significantly affect the fatty acid concentrations in the hypothalamus. Furthermore, there were no significant differences in mRNA expression of CRH, MR and GR in the hypothalamus or of GR mRNA expression in the spleen. Concerning the immune status, the plasma IgG levels tended to be higher in SBP pigs, whereas the leukocyte distribution, mitogen-stimulated lymphocyte proliferation, and serum IgM levels remained unchanged. In conclusion, the SBP supplementation of the diet only caused moderate effects on fatty acid metabolism, but no significant effects on hypothalamic-pituitary-adrenal (HPA) activity and immunity in growing pigs. It seems that a beneficial effect of dietary n-3 PUFA on health and welfare is more likely to be expected during stressful situations.

NEW APPROACH TO STRESS RESEARCH IN PHOCIDS-POTENTIAL OF DEHYDROEPIANDROSTERONE AND CORTISOL/DEHYDROEPIANDROSTERONE RATIO AS MARKERS FOR STRESS IN HARBOR SEALS ( PHOCA VITULINA) AND GRAY SEALS ( HALICHOERUS GRYPUS).

Cortisol is known to reflect the level of the stress response. However, measuring the cortisol concentration only once fails to provide sufficient information about the duration of the stress exposure. Moreover, handling is an acute stressor and increases cortisol secretion especially in wildlife species. Yet, in phocids reliable indicators are missing that reflect potential chronic effects of stress. The adrenal-derived steroid hormone dehydroepiandrosterone (DHEA) has been described as biomarker for the assessment of the stress status but has not been applied to marine mammal stress research. Therefore, DHEA, the sulfated precursor DHEAS and the cortisol/DHEA ratio were determined in serum of different seals. One group consisted of harbor ( Phoca vitulina) and gray seals ( Halichoerus grypus) that were habituated to human handling. The other two groups included healthy and free-ranging seals suffering from a disease. Blood samples were taken from 11 habituated (six males, five females), 17 wild_healthy (13 males, four females), and nine wild_diseased seals (four males, five females). No differences in serum cortisol levels could be detected between wild_healthy and wild_diseased seals. On the contrary, wild_diseased seals showed the lowest DHEA concentration compared to the other two groups. Wild_diseased seals also revealed the highest cortisol/DHEA ratio compared to the habituated_zoo (2,074.7 ± 351.4 vs. 121.5 ± 26.8, P < 0.001) and wild_healthy seals (827.9 ± 214.3, P < 0.01). DHEA and the cortisol/DHEA ratio may reflect differences in the functionality of the hypothalamic-pituitary-adrenal axis and therefore represent valuable tools for the assessment of stress-related effects in seals.

Oral Supplementation with a Special Additive of Retinyl Palmitate and Alpha Tocopherol Reduces Growth Retardation in Young Pancreatic Duct Ligated Pigs Used as a Model for Children Suffering from Exocrine Pancreatic Insufficiency.

Pancreatic exocrine insufficiency (PEI) is a disease of diverse aetiology-e.g., majority of patients suffering from cystic fibrosis (CF) show PEI congenitally. Malnutrition and malabsorption of nutrients impair growth and nutritional status. As reduced fat digestion leads to a deficiency of fat-soluble vitamins the supplementation is standard, but absorption is a critical point in PEI-patients. The pancreatic duct ligated (PL) pig is an established model for PEI in humans and has been proven to be a suitable model to compare different vitamin additives for supplementation. In a former study, PEI caused distinct growth retardation in young piglets, but did not affect growth in older ones. Our study hypothesised that this age-dependent effect is caused by exhausted body reserves of fat-soluble vitamins and, therefore, extra supply reduces growth retardation. PEI was induced by PL at the age of seven (PL-7) or 16 weeks (PL-16). Controls (C) underwent a sham surgery. Some PL-7 pigs (PL-7 + Vit) were fed a special vitamin additive. PEI reduced the mean final body weight (kg) at 26 weeks of age significantly with lower effect in PL-16-pigs (C:117; PL-7:49.5; PL-7 + Vit:77.1; PL-16:96.4). Extra vitamin supply resulted in an increased growth and normalised serum concentration of alpha-tocopherol, underlining the importance of special supplementation in PEI-patients.

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.

Effects of dietary nitrogen and/or phosphorus reduction on mineral homeostasis and regulatory mechanisms in young goats.

Introduction: The reduction of nitrogen (N) and phosphorus (P) in ruminant feed is desirable due to costs and negative environmental impact. Ruminants are able to utilize N and P through endogenous recycling, particularly in times of scarcity. When N and/or P were reduced, changes in mineral homeostasis associated with modulation of renal calcitriol metabolism occurred. The aim of this study was to investigate the potential effects of dietary N- and/or P-reduction on the regulatory mechanisms of mineral transport in the kidney and its hormonal regulation in young goats. Results: During N-reduction, calcium (Ca) and magnesium (Mg) concentrations in blood decreased, accompanied by a lower protein expression of cytochrome P450 family 27 subfamily B member 1 (CYP27B1) (p = 0.016). The P-reduced fed goats had low blood phosphate concentrations with simultaneously high Ca and Mg levels. The insulin-like growth factor 1 concentrations decreased significantly with P-reduction. Furthermore, gene expression of CYP27B1 (p < 0.001) and both gene (p = 0.025) and protein (p = 0.016) expression of the fibroblast growth factor receptor 1c isoform in the kidney were also significantly reduced during a P-reduced diet. ERK1/2 activation exhibited a trend toward reduction in P-reduced animals. Interestingly, calcitriol concentrations remained unaffected by either restriction individually, but interacted significantly with N and P (p = 0.014). Additionally, fibroblast growth factor 23 mRNA expression in bone decreased significantly with P-restriction (p < 0.001). Discussion: These results shed light on the complex metabolic and regulatory responses of mineral transport of young goats to dietary N and P restriction.

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.

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.

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.