The potential effects and interactions of oxidative stress and trace minerals on fresh and frozen semen in bulls - a review.

Reproduction is one of the most important factors determining successful cattle farming systems. Management practices, such as nutritional supplementation, can influence the reproductive performance of cattle. The objective of this literature review is to determine the potential value of injectable trace mineral administration on fresh and cryopreserved semen quality of bulls. A search of keywords related to the topic was performed on published articles and textbooks. The search was narrowed to the 40 most relevant references. Several studies have demonstrated a positive association between trace mineral supplementation and bull semen quality. Moderate amounts of reactive oxygen species (ROS) play an important role in normal spermatogenesis, but oxidative stress (OS), as experienced with adverse environmental conditions or disease, can contribute to idiopathic male infertility by negatively impacting spermatogenesis. Trace minerals such as selenium, copper, zinc, and manganese have been demonstrated to have antioxidant effects in mammals. Due to the complexity of oral ingested trace mineral bioavailability, injectable trace mineral supplementation prior to physiological periods with known deficiencies or increased requirement can benefit the animal. The potential benefits of injectable trace mineral supplementation to minimise oxidative damage to spermatogenesis in breeding bulls need further investigation. Positive results from such studies can lead to the implementation of injectable trace mineral supplementation strategies prior to the breeding season to minimise the detrimental effects of OS and can improve semen quality.

Anthelmintic resistance of gastrointestinal nematodes in dairy calves within a pasture-based production system of south West Western Australia.

The objective of this study was to determine the prevalence of gastrointestinal nematodes among post-weaned calves aged between 4 and 12 months old within a pasture-based system of south west Australia and quantify the level of anthelmintic resistance. Pre-treatment FECs were monitored on 14 dairy farms. Anthelmintic resistance was assessed on 11 of the farms. Control FECs were compared with anthelmintic FECs at 14 days post-treatment with doramectin (injectable), levamisole (oral), fenbendazole (oral) and a levamisole/abamectin combination (pour-on). Results demonstrate a strong level of anthelmintic resistance, with at least one class of anthelmintic failing to achieve a 95% reduction in FEC in one or more gastrointestinal nematode species. Doramectin was fully effective against Ostertagia, but C. oncophora displayed resistance in 91% of the farms. Conversely, levamisole was fully effective against C. oncophora, but Ostertagia displayed resistance in 80% of the farms. Fenbendazole resistance was present in both C. onocphora and Ostertagia in 64% and 70% of the farms, respectively. Trichostrongylus showed low resistance, occurring in doramectin (14%) and levamisole/abamectin combination (14%). This study confirms that anthelmintic resistance is common. Regular FEC reduction testing is recommended to monitor and guide decision-making for appropriate anthelmintic usage.

A pilot study on bacterial isolates associated with purulent vaginal discharge in dairy cows in the south-west region of Western Australia.

This study aimed to determine the bacterial isolates associated with postpartum endometritis among dairy cows in Western Australia and their antimicrobial susceptibility profiles. A cross-sectional study was conducted between June-October 2020. Endometritis was defined as evidence of mucopurulent to purulent vaginal discharge 60-100 days postpartum. Vaginal discharge samples were obtained, cultured, identified and tested for antimicrobial susceptibility. A total of 118 bacterial isolates were grown from 46 animals, representing 36 species. The bacteria isolated from both aerobic and anaerobic cultures included Bacillus (60.2%), Streptococcus (12.7%), Trueperella (10.1%), Escherichia (6.7%) and Staphylococcus (5.9%). The remaining genera <5% were Histophilus, Aeroccocus, Enterococcus and Moraxella. Resistance was variable between isolates, but the highest resistance levels were observed in Streptococcal and Bacillus isolates to enrofloxacin, clindamycin and erythromycin, respectively. All Streptococcal isolates exhibited 100% resistance to enrofloxacin, and the greatest resistance levels were found in Streptococcus luteinises to trimethoprim-sulfamethoxazole 83%, clindamycin 66% and 33% quinupristin-dalfopristin. There was 84.5% resistance to clindamycin and 35.2% to erythromycin in the Bacillus isolates, with the highest resistance found in Bacillus licheniformis and Bacillus subtilis. Escherichia coli exhibited 12.5% resistance to gentamycin, ceftiofur, whereas amoxicillin-clavulanic acid exhibited 37.5%. Within the Staphylococcal isolates, 28.5%, 28.5%, 42.8% and 14.2% resistance to ceftiofur, erythromycin, cefoxitin, penicillin and tetracycline were observed, respectively. The presence of resistance to important antimicrobials for human use, such as cephalosporins, macrolides and fluoroquinolones, highlights the need for judicious use of antimicrobials in dairy cattle.

Seminal transmission of lumpy skin disease virus in heifers.

It is known that lumpy skin disease virus (LSDV) can be shed in bull semen following infection and also that artificial insemination (AI) poses a biosecurity risk. However, it is not known whether the use of LSDV infected semen in AI poses a biosecurity risk. The aim of this study was to investigate whether LSDV, transmitted through semen, can infect cows and their embryos. Two controlled trials were performed simultaneously. Eleven young beef heifers, naïve to LSDV, were synchronized using an OvSynch protocol and inseminated on Day 0 with fresh semen spiked with a field strain of LSDV on day 0. Six of the heifers were superovulated on Day 1 using pregnant mare serum gonadotropin, and embryos were flushed from these heifers on Day 6. Blood and serum samples were collected from Day 4 until Day 27 to determine the presence of LSDV by PCR and virus isolation, and the presence of antibodies against LSDV by SNT. The first clinical signs of LSD were noticed on Day 10, followed by severe generalized LSD in three heifers and mild LSD in two more heifers. Two heifers were humanely euthanized due to severe unresponsive stranguria. LSDV was detected by PCR, virus isolation or electron microscopy in blood, embryos and organs of experimentally infected animals; and eight heifers had seroconverted by Day 27. Two control animals were not affected. This is the first report of experimental seminal transmission of LSDV in cattle.

Reversibility of the effects of GnRH-vaccination used to suppress reproductive function in mares.

REASONS FOR PERFORMING STUDY: Active immunisation against gonadotrophin-releasing hormone (GnRH) provides a reversible method for control of oestrous behaviour and fertility in mares. Previous reports failed to demonstrate the interval to resumption of cyclic ovarian activity after GnRH-vaccination. HYPOTHESIS: Administration of the GnRH-vaccine Improvac in a large group of mares of various ages will result in effective, reliably reversible suppression of ovarian activity within a 2 year period. METHODS: The mares, subdivided into 3 age categories, were vaccinated twice (with a 35 day interval) using 400 µg Improvac and monitored via blood samples until Day 720 after initial vaccination for serum progesterone concentration determination by radioimmune assay and anti-GnRH antibody titre by enzyme immunoassay. Samples were collected until individuals resumed cyclic ovarian activity. RESULTS: All mares showed suppression of cyclic ovarian activity by clinical examination and serum progesterone concentration (SPC) ≤ 1 nmol/l by Day 70 and 92.2% resumed cyclic activity by SPC at Day 720 with a mean interval = 417.8 days (s.d. ± 23.9; range 232-488 days, median 344 days). A significant age effect (P = 0.028) on the interval, but not on GnRH-antibody titre response, was observed between the youngest (≤ 4 years) and oldest (≥ 11 years) categories. CONCLUSIONS: Immunising adult mares of all ages with Improvac resulted in a reversible suppression of cyclic ovarian activity in most mares. An age effect, with the youngest mares showing a longer interval to reversibility, was observed.

Sites of persistence of lumpy skin disease virus in the genital tract of experimentally infected bulls.

The objectives of this work were to determine the site of persistence of lumpy skin disease virus (LSDV) in bulls shedding the virus in semen for a period longer than 28 days, to determine if the virus is present in all fractions of semen and to study lesions that developed in the genital tract. Six serologically negative postpubertal bulls were experimentally infected with a virulent field isolate of LSDV. The polymerase chain reaction (PCR) was performed on sheath washes, vesicular fluid, supernatant and cell-rich fractions of semen from day 10 to day 26 postinfection (p.i.). Bulls that were positive by PCR on the whole semen sample collected on day 28 p.i. were slaughtered and tissue samples from their genital tracts submitted for histopathological evaluation, immunoperoxidase staining, virus isolation and PCR. Two of the bulls developed severe lumpy skin disease (LSD) and were found to be shedding viral DNA in their semen on day 28 p.i. Viral DNA was identified in all semen fractions from all bulls, but mostly from the cell-rich fraction and from the severely affected bulls. The PCR assay was positive on postmortem samples of testes and epididymides from the two severely affected bulls. Virus could be recovered from the testes of these two bulls and from the epididymis of one of them. Immunoperoxidase staining was positive for LSDV staining in sections of testes and epididymides exhibiting necrosis. This study suggests that the testis and epididymis are sites of persistence of LSDV in bulls shedding virus in semen for prolonged periods and revealed that viral DNA is present in all fractions of the ejaculate.

The use of electrochemically activated saline as a uterine instillation in pony mares.

Twelve pony mares were randomly assigned to either a control or a treatment group and inseminated with fresh, raw semen from a single stallion of known fertility in a cross-over trial design. Pregnancy was diagnosed by transrectal ultrasound 12-14 days post-ovulation and then terminated by administration of a luteolytic dose of cloprostenol. Treatment mares received a uterine instillation of 100 ml of electrochemically activated (ECA) saline 4-12 hours post-insemination. Control mares received no treatment post-insemination. Per cycle pregnancy rate was 58.3 % in the control group and 50 % in the treatment group. There was no statistical difference (P = 1.000) in pregnancy rate between the 2 groups. The principles of ECA and applications of ECA saline are discussed.

Absence of lumpy skin disease virus in semen of vaccinated bulls following vaccination and subsequent experimental infection.

Twelve serologically negative bulls were used, six were vaccinated with a modified live LSD vaccine and six unvaccinated. All were then experimentally infected with a virulent field strain of LSDV. No clinical abnormality was detected following vaccination, and mild clinical signs were seen in four vaccinated bulls following challenge. Virus was not found in semen of vaccinated bulls. Two of the unvaccinated bulls developed severe LSD and four showed mild symptoms, all excreted the virus in the semen following challenge. This study confirmed the ability of LSD vaccination to prevent the excretion of LSDV in semen of vaccinated bulls.

Elimination of toxicity and enhanced detection of lumpy skin disease virus on cell culture from experimentally infected bovine semen samples.

Lumpy skin disease virus (LSDV), a poxvirus of the genus Capripoxvirus, is shed in the semen of infected bulls. The screening of semen for infectious virus requires a sensitive diagnostic method. The isolation of the virus on cell cultures and/or the polymerase chain reaction (PCR) are sensitive diagnostic tests which may be used to screen semen for LSD viral DNA prior to artificial insemination. Although cell culture detects infectious virus and is a sensitive method, there are major difficulties in using this method due to the toxic effect of semen on the cells. The aim of this study was to find a method that decreases the toxic effect of semen and enhances the isolation of LSDV on cell culture. Semen samples from LSDV sero-negative bulls were collected and infected with a field isolate of LSDV, strain V248/93, with a titre of 6.5 log TCID50. The semen samples were treated with one of four different methods: centrifugation, serial dilution, filtration and chemical treatment with kaolin. The samples subjected to centrifugation, serial dilution and filtration were supplemented with gentamycin. Semen toxicity on cell cultures was eliminated when supernatants of semen samples centrifuged at 2000 rpm for 1, 3 and 5 min and serially diluted were used to inoculate confluent monolayer bovine dermis cells. The toxicity recorded when the pellet fractions of semen samples centrifuged for 5 min at 2000 rpm was comparable to results obtained from serially diluted samples supplemented with gentamycin. Filtration and kaolin treatment of semen samples did not remove the toxic effect.