Temporal trends in bulk tank milk antibody ELISA and PCR test results for bovine viral diarrhoea in New Zealand pastoral dairy herds.

Aims: To describe temporal trends in bulk milk antibody ELISA and PCR testing for bovine viral diarrhoea (BVD) in New Zealand pastoral dairy herds and to assess the use of historical accession data to predict herd-level BVD incursions. Methods: Data on all diagnostic testing of bulk milk for BVD performed by the Livestock Improvement Corporation (Hamilton, NZ) over eight lactation seasons from 1 June 2010 to 31 May 2018 were analysed. This included anonymised herd identification, geographic location, herd size, sample collection date, sample to positive (S/P) ratio for antibody ELISA results, and cycle threshold values for PCR detecting viral RNA. Multivariable logistic regression was used to explore the relationship between historical accession data and the risk of herds having at least one positive bulk milk PCR test result in the 2017 season. Results: There were 156,034 bulk milk BVD diagnostic testing accessions for 10,495 uniquely identified dairy herds over the 8-season period. The prevalence of tested herds with at least one positive bulk milk PCR test result decreased from 14.6% (407/2,786) in the 2010 season to 5.6% (355/6,309) in the 2017 season with similarly marked declines in S/P ratios. In the 2017 season, 2,961/6,309 (46.9%) herds had S/P ratios greater than the 0.75 cut-off value indicating recent or active BVD virus transmission within the herd while 1,422/6,309 (22.5%) herds were classified as having negative or low S/P ratios. Herds that cleared BVD from the milking herd experienced a mean decline in S/P ratio of 0.11 units per year (min 0.05; max 0.18). In the multivariable analysis, the overall incidence risk of herds experiencing a BVD incursion in the 2017 season was 3.8% (146/3,848) and there were three significant predictors in the final model: herd size, PCR status in the 2014 season, and change in S/P ratio between the 2014 and 2015 seasons. The area under the receiver operating curve for the final model was 0.695 indicating poor discrimination. Conclusions and clinical relevance: The prevalence of dairy herds in New Zealand with positive bulk milk PCR test results and high S/P ratios has decreased over time, suggesting fewer herds are actively infected with BVD and that herd immunity may also be declining. Although monitoring trends in bulk milk test results provides useful information on changes in individual herd status, it is difficult to accurately predict when new incursions will occur and farmers should continue to maintain good biosecurity.

Use of an enzyme-linked immunosorbent assay for detecting bovine viral diarrhoea virus antibodies in individual cow milk samples.

AIMS: To calculate and verify an adjustment for the test result cut-off when testing milk samples using a bovine viral diarrhoea (BVD) antibody test so that it is a better approximation of the test on serum as the reference. Previous experience has demonstrated that different cut points may be required for results from milk and serum. METHODS: Paired milk and serum samples were collected from 90 cows from three herds in two regions of New Zealand and were tested using a commercially available bovine viral diarrhoea virus (BVD) antibody ELISA. A regression equation with second-order polynomial was developed to estimate a milk sample to positive-control optical density ratio (S/P ratio) value from a serum S/P ratio, and the milk values equivalent to the recommended serum cut points were calculated. The change in milk test result agreement with the result based on serum samples was assessed primarily using weighted Kappa statistics. RESULTS: The new cut points between negative and suspect, and suspect and positive milk samples were defined as S/P ratios of ≥ 0.08 and ≥ 0.13 respectively, compared to cut points for serum samples of ≥ 0.20 and ≥ 0.30, respectively. Using the new cut points increased the level of agreement (weighted Kappa) between serum and milk from 0.58 to 0.82. CLINICAL RELEVANCE: Milk BVD antibody testing can be used to estimate serum BVD antibody status. This is particularly useful for investigations where susceptibility to infection or the incidence of new infections is of interest. CONCLUSIONS: Using new adjusted cut points for milk, ELISA BVD antibody testing resulted in a higher level of agreement with serum samples.

Survey of bulk tank milk in New Zealand for Mycoplasma bovis , using species-specific nested PCR and culture.

AIM: To survey the dairy cattle population in New Zealand for the presence or absence of Mycoplasma bovis. METHODS: A random cross-sectional survey of bulk tank milk from dairy herds in New Zealand based on regionally proportioned sampling, weighted towards herds with a high bulk tank milk somatic cell count (SCC) was used to detect M. bovis at a between-herd prevalence of 2%, with 99% confidence. Bulk tank milk samples collected on-farm were tested using a nested M. bovis PCR, and bacteriological culture employing enrichment in mycoplasma broth and direct plating onto mycoplasma agar. RESULTS: Mycoplasma bovis was not detected in any of the 244 bulk tank milk samples by either PCR or culture. CONCLUSIONS: This survey provides further evidence that M. bovis is not present in the dairy cattle population in New Zealand.

Does control of bovine viral diarrhoea infection make economic sense?

AIM: To provide an economic analysis of the costs of control or eradication of bovine viral diarrhoea (BVD) against the estimated costs of the disease. METHODS: A decision-tree approach was adapted to an analysis of the costs of bovine viral diarrhoea virus (BVDV) infection and that of three main control options (vaccination, test-and-cull, and increased biosecurity) and their combinations, to the dairy industry in New Zealand. The model was based on an average herd of 322 milking cows. Endemic, epidemic and sporadic effects of BVDV infection were modelled in the herd, to derive an estimate of costs. RESULTS: The cost of BVDV infection to an infected average-sized dairy herd in New Zealand was estimated to be NZ $11,334 (or NZ $35.19 per cow) per annum, and NZ $48,311 over 10 years. Based on these calculations, the estimate of the annual cost of BVDV infection to the dairy industry in New Zealand was in excess of NZ $23 million per annum. While all of the control options required financial input, the rate of return compared with the cost of BVD, when viewed over a 10-year term, was as high as 123%. CONCLUSIONS: All control options offered considerable savings compared with the cost of BVD infection, and control is economically favourable. Uncertainty over the likely efficacy of the control options under field conditions in New Zealand would not allow a firm choice of one option over another at this stage, and more work on determining the efficacy of those control options in New Zealand is needed.

Insemination of susceptible heifers with semen from a non-viraemic bull with persistent bovine virus diarrhoea virus infection localized in the testes.

Bulls shedding bovine viral diarrhoea virus (BVDV) in semen and simultaneously having a high concentration of circulating antibodies may cause reproductive problems and spread the viral infection within cattle populations. To investigate this in detail, three heifers were inseminated with BVDV-infected semen from a non-viraemic, seropositive Holstein-Friesian bull, named 'Cumulus'. One control heifer was inseminated with semen from a healthy bull that was free of BVDV. All four heifers remained clinically healthy throughout the experiment. The conception succeeded in the control animal and in two of the three heifers inseminated with semen containing BVDV. The heifer with the failed conception was the only one that became systemically infected with BVDV. This animal was deemed non-pregnant by ultrasonic examination on day 34 after insemination and showed no signs of subsequent oestrus during the entire experimental period. At slaughter, 42 days after insemination, there were no histopathological changes in the ovaries and virus was not detected in ovarian tissue. The fact that seronegative dams served with semen from persistently infected bulls have occasionally produced persistently infected calves together with the present findings and the fact that non-viraemic, seropositive bulls can constantly shed BVDV, suggest that the use of semen from such bulls in BVDV-free herds could have far-reaching consequences, especially if it led to the birth of persistently infected (P1) calves.

Persistent bovine pestivirus infection localized in the testes of an immuno-competent, non-viraemic bull.

A post-pubertal bull on an artificial insemination station was found to be persistently shedding bovine viral diarrhoea virus (BVDV) in semen over a period of eleven months, while demonstrating no viraemia. Circulating antibodies to BVDV were consistently high, suggesting that the immune system was challenged repeatedly. Post-mortem findings confirmed that the virus was sequestered in the testes of the bull. It is hypothesized that the BVDV in this immuno-competent bull was protected from the bull's immune response by the blood-testes barrier. The barrier becomes functional only at puberty when tight junctions form between adjacent Sertoli cells, suggesting that this bull became persistently infected with BVDV during puberty.

Identification and imaging of OH (nu'' = O) and O(2) (nu'' = 6 or 7) in an automobile spark-ignition engine using a tunable KrF excimer laser.

Planar laser-induced predissociative fluorescence is applied to image state-specific densities of OH and hot O(2) inside an internal-combustion car engine. Improved instrumentation is described. It includes better imaging optics and a spectrometer that permits desired molecular quantum states to be selected and identified in real time. The OH (nu'' = 0) images are cleanly separated from the isooctane fuel and they display a thin superequilibrium region at the flame front. In contrast, vibrationally excited O(2) (nu'' = 6 or nu'' = 7) is uniformly distributed behind the front. Uneven and broken flame fronts are commonly observed.

Fluorescence imaging inside an internal combustion engine using tunable excimer lasers.

Tunable excimer lasers are used to obtain 2-D images of molecular (and some state-specific) density distributions inside a cylinder of a modified four-cylinder in-line engine that has optical access. Natural fluorescence (i.e., without a laser) is used for some OH pictures, normal laser-induced fluorescence (LIF) for those of NO and of the isooctane fuel, and laser-induced predissociative fluorescence (LIPF) for other OH pictures and for those of O(2). Relevant spectroscopy is done to find the laser and fluorescence frequencies needed to measure isolated species. LIPF works well at high pressures, is state specific, and is ideally suited to follow turbulent processes. No similar measurements in engines have been previously reported. Pictures are taken in succeeding engine cycles. Their sequence is either at a particular point of the engine's cycle to show cyclic fluctuations, or at succeeding portions of the cycle to illustrate the progress of the gasdynamics or of the combustion.

High-sensitivity detection of NO in a flame using a tunable ArF laser.

A tunable, narrow-band ArF laser has been used for laser-induced fluorescence detection of NO in natural abundance in a flame experiment. P and R branches of the D(2)Sigmaupsilon' = 0?X(2)Pi(3/2,1/2)upsilon'' transition were observed probing rotational states between J'' = 19.5 and J'' = 44.5. A single-shot detection limit of 1 part in 10(6) was found with a monochromator-based, dispersed-fluorescence detection system. In an experimental setup, determination of undispersed laser-induced fluorescence detection limits at or below the 1-part-in-10(9) range should be possible, because the narrow-band laser can be used to suppress all other sources of contaminating fluorescence even for detection of trace NO. The NO B(2)Piupsilon' = 7 ? X(2)Piupsilon'' = 0 transition was also observed in a cell experiment but not in the flame and is reported here.