Scrotal tick damage as a cause of infertility in communal bulls in Moretele, South Africa.

Calving rate in communal cattle influences both food security and socio-economics in rural households. A previous study indicated that scrotal damage caused by ticks could affect the fertility of communal bulls and reduce the annual calving rate. The objectives of the study were to investigate the annual calving rate in communal herds by counting calves during herd visits, perform breeding soundness examinations on bulls and identify adult ticks attached to their genitalia. This prospective longitudinal survey was based on participatory rural appraisal. Calving rates were estimated in cows (n = 2398) from 100 randomly selected communal herds in Moretele over 12 months in 2013, during routine visits by animal health technicians. Randomly selected bulls (n = 50) from these herds were tested for Brucella abortus, Trichomonas foetus and Campylobacter fetus subspecies venerealis. The calving rate was 35.86% (0.359). The mean scrotal circumference was 37.63 ± 3.42 cm. Total sperm motility was 78.73 ± 35.73%; progressive sperm motility was 27.39 ± 15.81% and non-progressive sperm motility was 51.34 ± 19.92%. Thirty-five of the 38 bulls examined for breeding soundness exhibited severe scrotal and preputial lesions caused by the adult ticks Amblyomma hebraeum and Hyalomma rufipes. Tick control methods used included spraying (n = 20), pour-on (n = 11), no control (n = 1) and various (n = 18). It was concluded that in Moretele genital tick damage had a more serious impact on the fertility of communal bulls than contagious diseases. Targeted acaricidal spot treatment of the genitalia of communal bulls to prevent infestation is recommended, as tick control strategies used by farmers appeared to be inadequate.

Effect of using frozen-thawed bovine semen contaminated with lumpy skin disease virus on in vitro embryo production.

Lumpy skin disease (LSD) is an important transboundary animal disease of cattle with significant economic impact because of the implications for international trade in live animals and animal products. LSD is caused by a Capripoxvirus, LSD virus (LSDV), and results in extensive hide and udder damage, fever and pneumonia. LSDV can be shed in semen of infected bulls for prolonged periods and transmitted venereally to cows at high doses. This study examined the effects of LSDV in frozen-thawed semen on in vitro embryo production parameters, including viral status of media and resulting embryos. Bovine oocytes were harvested from abattoir-collected ovaries and split into three experimental groups. After maturation, the oocytes were fertilized in vitro with frozen-thawed semen spiked with a high (HD) or a lower (LD) dose of LSDV, or with LSDV-free semen (control). Following day 7 and day 8 blastocyst evaluation, PCR and virus isolation were performed on all embryonic structures. After completing sufficient replicates to reach 1,000 inseminated oocytes, further in vitro fertilization (IVF) runs were performed to provide material for electron microscopy (EM) and embryo washing procedures. Overall, in vitro embryo yield was significantly reduced by the presence of LSDV in frozen-thawed semen, irrespective of viral dose. When semen with a lower viral dose was used, significantly lower oocyte cleavage rates were observed. LSDV could be detected in fertilization media and all embryo structures, when higher doses of LSDV were present in the frozen-thawed semen used for IVF. Electron microscopy demonstrated LSDV virions inside blastocysts. Following the International Embryo Transfer Society washing procedure resulted in embryos free of viral DNA; however, this may be attributable to a sampling dilution effect and should be interpreted with caution. Further research is required to better quantify the risk of LSDV transmission via assisted reproductive procedures.

Serum albumin concentration of donor cows as an indicator of developmental competence of oocytes.

Adequate nutrition is required for maintenance of normal reproduction in cattle. Albumin, the best marker and fundamental part of nutrition, most abundant plasma protein and major component of fetal bovine serum, is the best predictor of malnourishment in South African cattle. The aim of this study was to determine if serum albumin concentrations of donor cows predict the developmental competence of oocytes, and if additional protein supplementation of the in vitro culture media improves embryo outcomes in oocytes from cows with inadequate serum albumin concentrations. Oocytes (n = 1024) were recovered from donors with inadequate (≤35.9 g/L), or adequate serum albumin concentrations (≥36.0 g/L). Four hundred and sixty oocytes originated from cows with inadequate serum albumin and 564 from cows with adequate serum albumin. Oocytes of these cohorts were randomly allocated to a control and supplemented fetal bovine serum in vitro embryo culture protocol. Multiple linear, logistic and Poisson regression analyses were performed to estimate the effects of different covariates on linear, binary and count data respectively. Mixed effects Poisson regression was performed for the number of oocytes that developed into blastocysts by the seventh day of culture. Adequate serum albumin concentration of donor cows independently resulted in 46% increased blastocyst formation in the control protocol (P = 0.02). Although fetal bovine serum supplementation of the culture protocol did not affect blastocyst formation in oocytes originating from cows with inadequate serum albumin, it independently reduced blastocyst formation by 30% in oocytes originating from cows with adequate serum albumin (P = 0.02). Other independent predictors of blastocyst outcome included higher serum urea nitrogen, lower beta (ß)-hydroxybutyric acid concentrations and lower fat classification of donor cows. It is concluded that adequate serum albumin of donor cows is a significant predictor of developmental competence of oocytes, and that in vitro supplementation of fetal bovine serum does not improve developmental competence of oocytes and can lead to negative blastocyst outcomes. Further research is required to determine optimal protein supplementation for oocytes originating from inadequately nourished cows.

Effect of semen processing methods on lumpy skin disease virus status in cryopreserved bull semen.

Lumpy skin disease is an economically important disease of cattle, caused by the lumpy skin disease virus (LSDV; Capripoxvirus). It has a variable clinical appearance but, in severely affected animals, is associated with extensive skin damage, pneumonia and death. The LSDV can be found in the semen of infected bulls for prolonged periods of time, from where it can be transmitted by mating or artificial insemination and cause clinical disease in heifers and cows. In this study, an ejaculate was collected from a LSDV seronegative bull and confirmed free from LSDV DNA by PCR. The ejaculate was split into a control sample (C), a sample spiked with a 4 log TCID50 dose of an LSDV isolate (HD) and a 103 dilution of the virus suspension (ND) and frozen routinely. Two straws from each of the different semen treatment groups (HD, ND and C) were subsequently thawed and subjected to swim-up, single layer centrifugation, Percoll® density gradient and a Percoll® density gradient with added trypsin. For one set of straws, semen quality variables were recorded, and viral DNA status determined using PCR; the other set was used for positive staining electron microscopy. Samples determined to be positive for LSDV DNA by PCR were then subjected to virus isolation (VI). Complete elimination of LSDV from semen did not occur with use of any of the processing methods. Trypsin did reduce the viral load, and eliminated LSDV from the ND sample, but severely negatively influenced semen quality. The LSDV virions, as assessed by electron microscopy, were associated with the sperm plasma membrane. Further investigation is needed to establish the efficacy of immuno-extenders for rendering semen free from LSDV.

Standardization criteria for an ischemic surgical model of acute hepatic failure in pigs.

The establishment of a reliable large animal model of acute hepatic failure (AHF) is critical for the evaluation of supportive therapies such as bioartificial liver support systems (BALSS). Large animal surgical models bear some resemblance to the clinical syndrome of AHF in humans. However, these are inherently complex and are predisposed to inter-individual variation. The development of considerable skill is required and no precise standardization criteria for such models have been defined as yet. This study investigates a surgically induced ischemic model of AHF in 15 female Landrace pigs. A large set of systemic and biochemical variables were measured. The absolute values of systemic variables during the surgery and the rates of change of the biochemical variables following the surgery were correlated with the duration of survival of each animal. A multivariate prognosis was revealed, with several variables simultaneously determining survival. Those of importance in the surgical period included the duration of portal occlusion, the pulse rate during this time, the total blood lost, the mean arterial pressure and the body temperature after the procedure. In the post-surgical period the rates of change of blood ammonia, branch chain amino acids, haemoglobin, hematocrit, body temperature, total urinary excretion and mean blood pH all demonstrated some importance to survival. Based on the above and clinical experience, standardization criteria specific for this model were defined. Additional variables that posses value in AHF but demonstrate lower correlations with survival were also included, e.g. the prothrombin time, clotting factors, liver enzymes, potassium, bilirubin, creatinine and lactate. Valuable procedural information was generated, which stabilized the model, limited unnecessary blood sampling and cut laboratory costs. The criteria allow the early exclusion of compromized animals and the identification of those demonstrating characteristics predictive of longer survival prior to BALSS connection. This is likely to allow more accurate comparisons between small treatment and control groups. In effect, the above are the converse of the King's college criteria (for humans) as they are designed to exclude animals with prognoses that are insufficiently bleak or due to factors other than AHF. The statistical analysis employed also identified the variables that in future may allow the evaluation of prognosis in real-time.