Reactive oxygen species (ROS)-induced oxidative stress is well known to play a major role in male infertility. Sperm are sensitive to ROS damaging effects because as male germ cells form mature sperm they progressively lose the ability to repair DNA damage. However, how oxidative DNA lesions in sperm affect early embryonic development remains elusive. Using cattle as model, we show that fertilization using sperm exposed to oxidative stress caused a major developmental arrest at the time of embryonic genome activation. The levels of DNA damage response did not directly correlate with the degree of developmental defects. The early cellular response for DNA damage, γH2AX, is already present at high levels in zygotes that progress normally in development and did not significantly increase at the paternal genome containing oxidative DNA lesions. Moreover, XRCC1, a factor implicated in the last step of base excision repair (BER) pathway, was recruited to the damaged paternal genome, indicating that the maternal BER machinery can repair these DNA lesions induced in sperm. Remarkably, the paternal genome with oxidative DNA lesions showed an impairment of zygotic active DNA demethylation, a process that previous studies linked to BER. Quantitative immunofluorescence analysis and ultrasensitive LC–MS-based measurements revealed that oxidative DNA lesions in sperm impair active DNA demethylation at paternal pronuclei, without affecting 5-hydroxymethylcytosine (5hmC), a 5-methylcytosine modification that has been implicated in paternal active DNA demethylation in mouse zygotes. Thus, other 5hmC-independent processes are implicated in active DNA demethylation in bovine embryos. The recruitment of XRCC1 to damaged paternal pronuclei indicates that oxidative DNA lesions drive BER to repair DNA at the expense of DNA demethylation. Finally, this study highlighted striking differences in DNA methylation dynamics between bovine and mouse zygotes that will facilitate the understanding of the dynamics of DNA methylation in early development. The data demonstrate that oxidative stress in sperm has an impact not only on DNA integrity but also on the dynamics of epigenetic reprogramming, which may harm the paternal genetic and epigenetic contribution to the developing embryo and affect embryo development and embryo quality.

Background: Recumbent cows are a diagnostic challenge because of a wide range of differential diagnoses, which include trauma, neurological and metabolic disorders, malnutrition and mineral deficiencies. This case report describes recumbent suckler cows that presented as a herd problem. In addition to weakness due to inanition, Cu and Se deficiencies were considered as possible aetiologies of the recumbency. Furthermore, Trypanosoma (T.) theileri, a blood parasite of unknown importance in Germany, was detected in the blood of some cows. Case presentation: Three recumbent cows were referred to the Clinic for Ruminants and Swine, Faculty of Veterinary Medicine of the University of Leipzig. They were unable to rise and had low body condition scores and rough hair coats. Haematological and serum biochemical analyses showed neutrophilia, electrolyte imbalances, increased activities of muscle and liver enzymes and decreased concentrations of trace elements, especially Copper (Cu) and Selenium (Se). T. theileri was detected in a routine blood smear from one cow. The cows did not respond to an intensive care protocol, which included intravenous fluids and electrolytes, mineral substitution, non-steroidal anti-inflammatories and antibiotics, and were therefore euthanized or died. Postmortem examination showed cachexia, subcutaneous and scleral oedema and muscular dystrophy, especially in the hind limbs. Follow-up examination of the herd of origin produced similar findings including the detection of T. theileri in a large proportion of the herd. Ration analysis revealed considerable undersupply of several nutrients. Conclusions: Based on all findings, an aetiological diagnosis of trace mineral and nutrient deficiency with possible involvement of T. theileri was made.

The extent of oxidative damage transferred by the damaged sperm to the progeny is likely to be limited by the oocyte’s repair and antioxidative capacity. We aimed to assess the association between Brilliant Cresyl Blue (BCB) staining in oocytes and their competence for embryo development after in vitro fertilisation (IVF) with damaged sperm. For this purpose, bovine sperm were incubated without (non-oxidised sperm, NOX S) or with 100 µM H₂O₂ (oxidised sperm, OX S) and were used to fertilise in-vitro-matured bovine oocytes (BCB-pos./BCB-neg.). Unstained oocytes served as controls (US). Development was assessed at 30, 46, 60 h and on Days (D) 7 and 8 after IVF. Total cell number and apoptotic index were analysed in D7 blastocysts. BCB-neg. oocytes showed lower cleavage rates and blastocyst rates than unstained oocytes after IVF with NOX S (p < 0.05). They showed the highest reduction in D7 blastocyst rate upon fertilisation with OX S and showed a delayed embryo development at 46 and 60 h after IVF compared to embryos produced with NOX S (p < 0.05). Total cell number in blastocysts produced with BCB-neg. oocytes was lower (p < 0.05) in the embryos produced with OX S than in embryos after IVF with NOX S. In conclusion, BCB-neg. oocytes have a lower competence to support embryo development after in vitro fertilisation with oxidised sperm.

Animal experimentation is required to investigate complex physiological relationships and facilitates development of evidence-based knowledge. However, experimental protocols can interfere with the daily routine of the animals, result in stress and pain, and have adverse effects on health and production. The goal of this study was to investigate the effects of an intensive experimental protocol on health traits and production in transition dairy cows. Eighty experimental dairy Holstein cows (EXP group) underwent serial protocol-based clinical and ultrasonographic examinations, puncture of the jugular vein for blood collection or drug application, and liver biopsy samples, 14 d before until 42 d after parturition. Controls (CTR group) included 206 cows from the same herd, which fulfilled the same inclusion criteria and were kept under the same production management but were not handled for the purpose of this study. Procedure-related effects with a potentially negative effect on health and production were recorded. Furthermore, production, fertility and culling traits of the 2 groups (CTR, EXP) were compared. Most procedure-related adverse effects were associated with transcutaneous liver biopsies and included diffuse inflammation of the skin incision in 11.9% (42 of 320), abscessation of the skin or subcutis in 4.6% (11 of 240), and increased liver echogenicity of the biopsy site in 10.4% (27 of 240). The experimental procedures had a negative effect on milk yield at the start [days in milk (DIM) 5-50, difference: 2.3 kg, standard error (SE): 0.8 kg] and end of lactation (DIM 251-300, difference: 2.0 kg, SE: 1.0 kg; DIM 301-350, difference: 2.3 kg, SE: 1.2 kg) resulting in a lower 305-d milk yield in the EXP group than in the CTR group (difference: 472 kg, SE 214 kg). The milk fat % was higher in the EXP group than in the CTR group from 251 DIM onward (DIM 251-300, difference: 0.20%, SE: 0.09%; DIM 301-350, difference: 0.41%, SE: 0.17%). Also, the somatic cell score was higher in the EXP group than in the CTR group, during early (5-50 DIM, difference: 0.43, SE: 0.22) and from mid-lactation onward (DIM 151-200, difference: 0.43, SE: 0.2; DIM 201-250, difference: 0.49, SE: 0.22; DIM 251-300, difference: 0.55, SE: 0.25; DIM 301-350, difference: 0.61, SE: 0.28). Experimental procedures had no effect on first service conception rate and time to pregnancy, but had a positive effect on stillbirth rate with fewer stillbirths in the CTR group (0%) than in the EXP group (3.9%). Furthermore, experimental handling had no effect on time to culling or type of culling, whereby poor production was a more frequent reason noted for culling in the EXP group. Procedure-associated impairment of production in dairy cows is rarely reported and allows the estimation of the effects of such a study protocol on animal health and production. As a limitation for the interpretation of the results, the number of animals included and conduction in one single herd have to be considered.

Bovine veterinarians are regularly confronted with teat lesions in cows. The number of studies on the diagnosis and treatment of teat lesions as well as the exchange of practical experience among clinicians are extensive in dairy cows compared with suckler cows. The aim of this case report was to describe the successful treatment of teat stenosis in a suckler cow and discuss possible challenges. A four-year-old Simmental cow, in her third lactation and 4 days in milk, was referred to our clinic along with her calf because of teat stenosis in the front left quarter. The owner had repeatedly used a rigid teat cannula in an attempt to relieve the stenosis during the previous lactation. However, the cow had refused to allow the current calf to suckle the affected teat and resisted attempts by the owner to cannulate the teat. The results of clinical examination, ultrasonography, and milk sampling showed stenosis of the proximal, middle, and distal parts of the front left teat cistern, accompanied by thelitis and cisternitis and mild chronic clinical mastitis. Based on published recommendations, treatment of the thelitis, cisternitis, and mastitis was initiated before resolution of the stenosis surgically. The first week of treatment included the administration of an intramammary product containing cefapirin and prednisolone, a systemic non-steroidal anti-inflammatory drug, a wax teat-boogie, and bandaging of the teat. Thereafter, the treatment was reduced to insertion of a wax-teat boogie and bandaging. Conservative treatment resulted in resolution of the mastitis, cisternitis, and stenosis in the proximal and middle parts of the teat, which had most likely been caused by repeated cannulation of the teat by the owner. Lateral theloscopy was then used to remove the distal stenosis, which was the primary lesion. Healing of the surgical wound and resolution of the swelling occurred several days postoperatively, and the calf’s first attempt to suckle the teat was successful. The cow and calf were discharged from the clinic 2 weeks after surgery. A follow-up visit 4.5 months after surgery revealed that the calf was still nursing the teat and the operated quarter was producing a normal amount of milk.