A sandwich enzyme immunoassay for bovine interferon-γ and its use for the detection of tuberculosis in cattle

An in vitro cellular assay for bovine tuberculosis has recently been developed. This assay detects gamma-interferon released in response to specific antigen in a whole blood culture system. The bio-assay previously described for the detection of bovine gamma-interferon (IFN-gamma) has now been replaced with a sandwich enzyme immunoassay (EIA) which utilises two monoclonal antibodies to bovine IFN-gamma. The EIA detects less than 25pg/ml of recombinant bovine IFN-gamma and is specific for biologically active bovine IFN-gamma; and does not detect bovine alpha or beta interferon. IFN-gamma from sheep, goat and buffalo, but not from pig, deer or man, are also recognised by the EIA. The bovine IFN-gamma EIA when used in conjunction with the whole blood culture system has resulted in a simple, rapid and sensitive in vitro assay for specific cell mediated immune responsiveness to M. bovis infection in cattle.     

Antibodies to iron-regulated outer membrane proteins of coliform bacteria isolated from bovine intramammary infections

Expression of iron-regulated outer membrane proteins (OMP) by Escherichia coli and Klebsiella pneumoniae initially isolated from bovine intramammary infections (IMI) was investigated. Additionally, the presence of antibodies in bovine serum and mammary secretion directed against the iron-regulated OMP was examined. Outer membrane proteins were separated by sodium-dodecyl polyacrylamide electrophoresis. Detection of immunoglobulin G directed against OMP was by immunoblotting. All Gram-negative bacteria expressed iron-regulated OMP when grown in skim milk or trypticase soy broth plus iron chelator, alpha-alpha'-dipyridyl. Immunoglobulin G directed against the iron-regulated OMP, as well as the major OMP and several other proteins, was detected in serum and milk of lactating cows with or without Gram-negative bacterial IMI. Antibody against the iron-regulated OMP was detected also in colostrum, secretion from the involuted gland, and in newborn calf serum 4 days after ingesting colostrum.     

Simulated rainfall evaluation of revegetation/mulch erosion control in the Lake Tahoe basin: 2. Bare soil assessment

Slopes that have been disturbed through roadway, ski slope or other construction often produce more sediment than less disturbed sites. Reduction or elimination of sediment loading from such disturbed slopes to adjacent streams is critical in the Lake Tahoe basin. Here, use of a portable rainfall simulator (RS), described in the first paper of this series, is used to evaluate slope effects on erosion from bare volcanic and granitic soils (road cut and ski run sites) common in the basin in order to establish a basis upon which revegetation treatment comparisons can be made. Rainfall simulations (60 mm h⁻¹, approximating a 100‐year, 15‐minute storm) at each site included multiple replications of bare soil plots as well as some adjacent ‘native’, or relatively undisturbed soils below trees where available. Field measurements of time to runoff, infiltration, runoff, sediment discharge rates, and average sediment concentration were obtained. Laboratory measurements of particle‐size distributions using sieve and laser counting methods indicated that the granitic soils had larger grain sizes than the volcanic soils and that road cut soils of either type also had larger grain sizes than their ski run counterparts. Particle‐size‐distribution‐based estimates of saturated hydraulic conductivity were 5–10 times greater than RS‐determined steady infiltration rates. RS‐measured infiltration rates were similar, ranging from 33–50 mm h⁻¹ for disturbed volcanic soils and 33–60 mm h⁻¹ for disturbed granitic soils. RS‐measured runoff rates and sediment yields from the bare soils were significantly correlated with plot slope with the exception of volcanic road cuts due to the narrow range of road cut slopes encountered. Sediment yields from bare granitic soils at slopes of 28 to 78 per cent ranged from ∼1 to 12 g m⁻² mm⁻¹, respectively, while from bare volcanic soils at slopes of 22 to 61 per cent they ranged from ∼3 to 31 g m⁻² mm⁻¹, respectively. Surface roughness did not correlate with runoff or erosion parameters, perhaps also as a result of a relatively narrow range of roughness values. The volcanic ski run soils and both types of road cut soils exhibited nearly an order of magnitude greater sediment yield than that from the corresponding native, relatively undisturbed, sites. Similarly, the granitic ski run soils produced nearly four‐times greater sediment concentration than the undisturbed areas. A possible goal of restoration/erosion control efforts could be recreation of ‘native’‐like soil conditions. Copyright © 2005 John Wiley & Sons, Ltd.     

Simulated rainfall evaluation of revegetation/mulch erosion control in the Lake Tahoe Basin—1: method assessment

Revegetation of road cuts and fills is intended to stabilize those drastically disturbed areas so that sediment is not transported to adjacent waterways. Sediment has resulted in water quality degradation, an extremely critical issue in the Lake Tahoe Basin. Many revegetation efforts in this semiarid, subalpine environment have resulted in low levels of plant cover, thus failing to meet project goals. Further, no adequate physical method of assessing project effectiveness has been developed, relative to runoff or sediment movement. This paper describes the use of a portable rainfall simulator (RS) to conduct a preliminary assessment of the effectiveness of a variety of erosion‐control treatments and treatment effects on hydrologic parameters and erosion. The particular goal of this paper is to determine whether the RS method can measure revegetation treatment effects on infiltration and erosion. The RS‐plot studies were used to determine slope, cover (mulch and vegetation) and surface roughness effects on infiltration, runoff and erosion rates at several roadcuts across the basin. A rainfall rate of ≈60 mm h⁻¹, approximating the 100‐yr, 15‐min design storm, was applied over replicated 0·64 m² plots in each treatment type and over bare‐soil plots for comparison. Simulated rainfall had a mean drop size of ≈2·1 mm and approximately 70% of ‘natural’ kinetic energy. Measured parameters included time to runoff, infiltration, runoff/infiltration rate, sediment discharge rate and average sediment concentration as well as analysis of total Kjeldahl nitrogen (TKN) and dissolved phosphorus (TDP) from filtered (0·45 μm) runoff samples. Runoff rates, sediment concentrations and yields were greater from volcanic soils as compared to that from granitic soils for nearly all cover conditions. For example, bare soil sediment yields from volcanic soils ranged from 2–12 as compared to 0·3–3 g m⁻² mm⁻¹ for granitic soils. Pine‐needle mulch cover treatments substantially reduced sediment yields from all plots. Plot microtopography or roughness and cross‐slope had no effect on sediment concentrations in runoff or sediment yield. RS measurements showed discernible differences in runoff, infiltration, and sediment yields between treatments. Runoff nutrient concentrations were not distinguishable from that in the rainwater used. Copyright © 2004 John Wiley & Sons, Ltd.