Maple syrup urine disease in calves: a clinical, pathological and biochemical study

The clinical, pathological and biochemical findings of a study of 30 Poll Hereford, Hereford, Poll Hereford cross or Hereford cross calves affected with branched chain ketoacid dehydrogenase (BCKAD) complex deficiency or maple syrup urine disease (MSUD) are presented. In breeding studies, 6 of 21 calves from obligate heterozygote matings were affected with MSUD, suggesting the disease is inherited in an autosomal recessive manner. Calves were clinically affected from birth, but there were variations in the subsequent course of progressive deterioration of central nervous system function. Concentrations of the branched chain amino acids and keto acids were elevated in pre-suckle plasma and cerebellar water content was higher in affected calves. Activity of BCKAD complex was minimal in fibroblasts cultured from an affected calf. Spongiform encephalopathy and elevated ratios of the branched to straight chain amino acids in formalin fixed cerebral tissue were found in a stillborn foetus and a 3-month-old Hereford calf. These findings suggest the disease occurs prenatally and that a delayed form may exist.     

Numbers and percent of T lymphocytes in bovine peripheral blood during the periparturient period.

To determine if periparturient immunosuppression in dairy cattle might be due to an alteration in total numbers of percent of T lymphocytes, we examined the numbers and percent of T lymphocyte subsets in peripheral blood from periparturient dairy cows, some of which received recombinant bovine granulocyte colony stimulating factor (rbG-CSF) during the study. Beginning 2 weeks preparatum through 4 weeks postpartum, peripheral blood mononuclear cells (PBMC) were collected and labeled with monoclonal antibodies to BoCD5, BoCD4, and BoCD8, and the percent of cells positive for each marker measured by flow cytometry. The percent of PBMC expressing BoCD5 (total T cells), and BoCD8 (T suppressor/cytotoxic cells) was not significantly different between the groups, or at different times before and after calving. The percent of PBMC expressing BoCD4 (T helper cells) was not significantly different between the groups, however, within both groups there was a higher percent of BoCD4+ cells after calving than during the prepartum period. In cows receiving rbG-CSF, total numbers of PBMC were significantly increased compared to controls during the postpartum treatment period.     

Expression, purification, and in vitro biological activities of recombinant bovine granulocyte-colony stimulating factor

Neutrophils are essential components of the innate immune system and they play a critical role in the defense of host against bacterial and fungal infections. The colony stimulating factors are a class of glycoproteins that are required for proliferation, differentiation, and functional activation of hematopoietic progenitor cells. Granulocyte-colony stimulating factor (G-CSF) is a member of this regulatory family of cytokines that specifically stimulates proliferation and maturation of precursor cells in the bone marrow into fully differentiated and functional neutrophils. G-CSF also modulates the biological activities of mature neutrophils in circulation. A bovine G-CSF (bG-CSF) cDNA clone (previously isolated and sequenced in our laboratory) was expressed in Escherichia coli and the biological activities of the solubilized protein from purified inclusion bodies were examined. Flow cytometric analysis of membrane antigen density of neutrophils activated with bG-CSF revealed an upregulation in the expression of CD11a (>114%), CD11b (>148%), CD11c (>87%), and CD18 (>109%). Expression of L-selectin was decreased by more than 43%. There was no change, however, in the expression of CD14. These findings indicate that recombinant bG-CSF (rbG-CSF) expressed in E. coli is biologically active and exerts the same type of effects on neutrophils in vitro as those of human G-CSF (hG-CSF).     

Comparison of carbon-stock changes,eddycovariance carbon fluxes and model estimates in coastal Douglas-fir stands in British Columbia

Background: The global network of eddy-covariance(EC) flux-towers has improved the understanding of the terrestrial carbon(C) cycle, however, the network has a relatively limited spatial extent compared to forest inventory data and plots. Developing methods to use inventory-based and EC flux measurements together with modeling approaches is necessary evaluate forest C dynamics across broad spatial extents.Methods: Changes in C stock change(ΔC) were computed based on repeated measurements of forest inventory plots and compared with separate measurements of cumulative net ecosystem productivity(ΣNEP) over four years(2003 – 2006) for Douglas-fir(Pseudotsuga menziesii var menziesii) dominated regeneration(HDF00), juvenile(HDF88and HDF90) and near-rotation(DF49) aged stands(6, 18, 20, 57 years old in 2006, respectively) in coastal British Columbia. ΔC was determined from forest inventory plot data alone, and in a hybrid approach using inventory data along with litter fall data and published decay equations to determine the change in detrital pools. These ΔC-based estimates were then compared with ΣNEP measured at an eddy-covariance flux-tower(EC-flux) and modelled by the Carbon Budget Model- Canadian Forest Sector(CBM-CFS3) using historic forest inventory and forest disturbance data.Footprint analysis was used with remote sensing, soils and topography data to evaluate how well the inventory plots represented the range of stand conditions within the area of the flux-tower footprint and to spatially scale the plot data to the area of the EC-flux and model based estimates.Results: The closest convergence among methods was for the juvenile stands while the largest divergences were for the regenerating clearcut, followed by the near-rotation stand. At the regenerating clearcut, footprint weighting of CBM-CFS3 ΣNEP increased convergence with EC flux ΣNEP, but not for ΔC. While spatial scaling and footprint weighting did not increase convergence for ΔC, they did provide confidence that the sample plots represented site conditions as measured by the EC tower.Conclusions: Methods to use inventory and EC flux measurements together with modeling approaches are necessary to understand forest C dynamics across broad spatial extents. Each approach has advantages and limitations that need to be considered for investigations at varying spatial and temporal scales.     

Identification of Apoptotic Bodies in Equine Semen

Apoptosis in the testis is required to ensure an efficient spermatogenesis. However, sometimes, defective germ cells that are marked for elimination during this process escape elimination in the testes, giving rise to ejaculates with increased percentages of abnormal and apoptotic spermatozoa and a high percentage of apoptotic bodies. Apoptosis markers in the ejaculate have been associated with low fertility, either in animals or humans. Therefore, the goal of this study was to investigate whether fresh equine semen contains apoptotic bodies [initially named Merocyanine 540 (M540) bodies] and to study the relationship between the quantity of these bodies and cell concentration, the volume of ejaculate, viability and motility. Moreover, we also studied whether the presence apoptotic bodies in fresh semen was related to the resistance of the stallion spermatozoa to being incubated at 37°C or being frozen and thawed. Fresh equine semen was stained with fluorescent dyes such as M540 and Annexin‐V. Active Caspase 3 was studied in fresh semen through Western blotting and immunofluorescence with a specific antibody. Sperm kinematics was assessed in fresh, incubated and thawed samples using computer‐assisted semen analysis, and viability was evaluated with the LIVE/DEAD Sperm Viability Kit. Overall, our results demonstrate for the first time the presence of apoptotic bodies in equine semen. The quantity of apoptotic bodies was highly variable among stallions and was positively correlated with Caspase 3 activity in fresh samples and negatively correlated with the viability and motility of stallion spermatozoa after the cryopreservation process.     

Anti‐Mullerian Hormone Concentration and Antral Ovarian Follicle Population in Murrah Heifers Compared to Holstein and Gyr Kept Under the Same Management

This study was performed to evaluate plasma concentrations of anti‐Mullerian hormone (AMH) and the ovarian antral follicle population (AFP) in different genetic groups. Cyclic heifers (13 Bubalus bubalis [Murrah]; 15 Bos taurus [Holstein] and 10 Bos indicus [Gyr]) were maintained under the same management and were synchronized with two doses of 150 μg IM d‐cloprostenol administered 14 days apart. After the second d‐cloprostenol treatment, heifers had their ovaries scanned daily by ultrasound to define the day of ovulation. On the same day, the AFP was determined and a plasma sample was collected to measure AMH. Murrah heifers had less AFP (25.6 ± 2.1 follicles; p = 0.01) and plasma AMH concentration (0.18 ± 0.03 ng/ml; p < 0.001) than Gyr (60.0 ± 12.2 follicles and 0.60 ± 0.12 ng/ml of AMH); however, data were similar when compared to Holstein (35.9 ± 6.8 follicles and 0.24 ± 0.06 ng/ml of AMH) heifers. Regardless of genetic background, there was a positive relationship between the AFP and plasmatic AMH concentration (Murrah [r = 0.62; p < 0.01], Holstein [r = 0.66; p < 0.001] and Gyr [r = 0.88; p < 0.001]). Also, when heifers were classified according to high‐ or low‐AMH concentration based on the average within each genetic group, high‐AMH heifers had greater (p < 0.0001) AFP than low‐AMH heifers. In conclusion, both Murrah and Holstein heifers presented lower plasma AMH concentration and AFP when compared to Gyr.     

Reconstruction of Current Flow and Imaging of Current-Limiting Defects in Polycrystalline Superconducting Films

Magneto-optical imaging was used to visualize the inhomogeneous penetration of magnetic flux into polycrystalline TlBa2Ca2Cu3Ox films with high critical current densities, to reconstruct the local two-dimensional supercurrent flow patterns and to correlate inhomogeneities in this flow with the local crystallographic misorientation. The films have almost perfect c-axis alignment and considerable local a- and b-axis texture because the grains tend to form colonies with only slightly misaligned a and b axes. Current flows freely over these low-angle grain boundaries but is strongly reduced at intermittent colony boundaries of high misorientation. The local (<10-micrometer scale) critical current density Jc varies widely, being up to 10 times as great as the transport Jc (scale of approximately 1 millimeter), which itself varies by a factor of about 5 in different sections of the film. The combined experiments show that the magnitude of the transport Jc is largely determined by a few high-angle boundaries.     

Effects of colostral antibody on susceptibility of calves to Cryptosporidium parvum infection

Effects of colostral antibody on susceptibility of calves to Cryptosporidium parvum infection were examined. Six calves were fed pooled colostrum that contained C parvum antibody, 6 times daily (at 4-hour intervals) for 7 days and then milk replacer for 7 days. Colostrum was obtained from healthy cows or cows inoculated parenterally with C parvum oocysts before parturition. Antibody content was determined in serum and colostrum whey, using an ELISA for anticryptosporidia immunoglobulin. Six calves were fed colostrum from healthy cows 1 time, and then milk replacer 6 times daily for 14 days. On day 1, all calves were challenge exposed with C parvum, PO, and were monitored daily for diarrhea and oocyst shedding. Bovine colostrum containing specific antibody to C parvum, at ELISA titers up to 10,240, was not effective in protecting calves against challenge exposure to C parvum.