Estimation of sensitivity and specificity of pregnancy diagnosis using transrectal ultrasonography and ELISA for pregnancy-associated glycoprotein in dairy cows using a Bayesian latent class model

AIMS: To determine the sensitivity (Se) and specificity (Sp) of pregnancy diagnosis using transrectal ultrasonography and an ELISA for pregnancy-associated glycoprotein (PAG) in milk, in lactating dairy cows in seasonally calving herds approximately 85–100 days after the start of the herd’s breeding period.

METHODS: Paired results were used from pregnancy diagnosis using transrectal ultrasonography and ELISA for PAG in milk carried out approximately 85 and 100 days after the start of the breeding period, respectively, from 879 cows from four herds in Victoria, Australia. A Bayesian latent class model was used to estimate the proportion of cows pregnant, the Se and Sp of each test, and covariances between test results in pregnant and non-pregnant cows. Prior probability estimates were defined using beta distributions for the expected proportion of cows pregnant, Se and Sp for each test, and covariances between tests. Markov Chain Monte Carlo iterations identified posterior distributions for each of the unknown variables. Posterior distributions for each parameter were described using medians and 95% probability (i.e. credible) intervals (PrI). The posterior median estimates for Se and Sp for each test were used to estimate positive predictive and negative predictive values across a range of pregnancy proportions.

RESULTS: The estimate for proportion pregnant was 0.524 (95% PrI?=?0.485–0.562). For pregnancy diagnosis using transrectal ultrasonography, Se and Sp were 0.939 (95% PrI?=?0.890–0.974) and 0.943 (95% PrI?=?0.885–0.984), respectively; for ELISA, Se and Sp were 0.963 (95% PrI?=?0.919–0.990) and 0.870 (95% PrI?=?0.806–0.931), respectively. The estimated covariance between test results was 0.033 (95% PrI?=?0.008–0.046) and 0.035 (95% PrI?=?0.018–0.078) for pregnant and non-pregnant cows, respectively. Pregnancy diagnosis results using transrectal ultrasonography had a higher positive predictive value but lower negative predictive value than results from the ELISA across the range of pregnancy proportions assessed.

CONCLUSIONS AND CLINICAL RELEVANCE: Pregnancy diagnosis using transrectal ultrasonography and ELISA for PAG in milk had similar Se but differed in predictive values. Pregnancy diagnosis in seasonally calving herds around 85–100 days after the start of the breeding period using the ELISA is expected to result in a higher negative predictive value but lower positive predictive value than pregnancy diagnosis using transrectal ultrasonography. Thus, with the ELISA, a higher proportion of the cows with negative results will be non-pregnant, relative to results from transrectal ultrasonography, but a lower proportion of cows with positive results will be pregnant.     

Seroprevalence in Household Raised Pigs Indicate High Exposure to GII Noroviruses in Rural Nicaragua

Information about porcine norovirus (PoNoV), genetically similar to human NoV (HuNoV), is limited from rural areas where household‐raised pigs are heavily exposed to faecal material which could facilitate transmission. Histo‐blood group antigens (HBGAs) are known susceptibility factors to NoV in humans and in a germfree piglet model but their role in susceptibility in the porcine population remains unknown. This study reports: (i) the seroprevalence and antibody titres to human norovirus (NoV) VLPs in household raised pigs; (ii) the distribution of HBGAs in relation to NoV IgG antibody titres and further characterization by blocking of GII.4 VLP binding to pig gastric mucins (PGM). The majority of pigs were seropositive to all three VLPs tested (58–70%) with seropositivity and cross‐reactivity increasing significantly with age. However, pig sera could not block the binding of NoV GII.4 VLPs (Dijon) to PGM suggesting no previous infection with this genotype. The majority of the pigs were H‐positive (84%), a susceptibility factor for human infections. IgG antibody titres were however higher in H‐negative (GMT = 247) as compared with H‐positive (GMT = 57) pigs, but after age stratification, this difference in antibody titres was only observed in pigs ≤1 month of age. In conclusion, serological data show that the porcine population of Nicaragua is highly exposed to NoV infections, and the association to HBGAs warrants further investigation.     

Flow Cytometric Chromosomal Sex Sorting of Stallion Spermatozoa Induces Oxidative Stress on Mitochondria and Genomic DNA

To date, the only repeatable method to select spermatozoa for chromosomal sex is the Beltsville sorting technology using flow cytometry. Improvement of this technology in the equine species requires increasing awareness of the modifications that the sorting procedure induces on sperm intactness. Oxidative stress is regarded as the major damaging phenomenon, and increasing evidence regards handling of spermatozoa – including sex sorting – as basic ground for oxidative damage. The aim of this study was to disclose whether the flow cytometric sorting procedure increases the production of reactive oxygen species (ROS), and to identify if ROS production relates to DNA damage in sorted spermatozoa using specific flow cytometry‐based assays. After sorting, oxidative stress increased from 26% to 33% in pre‐ and post‐incubation controls, to 46% after sex sorting (p < 0.05). Proportions of DNA fragmentation index post‐sorting were approximately 10% higher (31.3%); an effect apparently conduced via oxidative DNA damage as revealed by the oxyDNA assay. The probable origin of this increased oxidative stress owes the removal of enough seminal plasma due to the unphysiological sperm extension, alongside a deleterious effect of high pressure on mitochondria during the sorting procedure.     

The Effect of Oxidative Stress on Thawed Bulk‐Sorted Red Deer Sperm

The aims of this study were to assess the effects of the sex‐sorting process on post‐thaw sperm quality as well as on induced oxidative stress damage (H₂O₂ 0 mm  = H000; H₂O₂ 50 mm  = H050; H₂O₂ 100 mm  = H100) and the protective action of reduced glutathione (GSH) and Trolox, when comparing sorted (BSS) and non‐sorted (NS) red deer spermatozoa incubated at 37°C. Sperm samples from three stags were collected by electroejaculation and frozen. Immediately after thawing, sperm motility was higher (p < 0.05) for NS (59% ± 3.3) than BSS (36.9% ± 5.8) sperm. Furthermore, the percentage of apoptotic sperm was higher (p < 0.05) for BSS (21.6% ± 5.0) than NS sperm (14.6% ± 1.2). The presence of H₂O₂ increased DNA damage in NS (H000 = 4.1% ± 0.9; H050 = 9.3% ± 0.7; and H100 = 10.9% ± 2.3), but not in BSS sperm. However, in the presence of oxidant, GSH addition improved (p < 0.05) sperm motility in both groups of sperm samples as compared to their controls (NS: 44.5 ± 4.8 vs 21.1 ± 3.9 and BSS: 33.3 ± 8.1 vs 8.9 ± 1.8). These results demonstrate that the sperm‐sorting process induces sublethal effects, albeit selecting a sperm population with a chromatin more resistant to oxidative stress than that in non‐sorted sperm. Moreover, addition of GSH at 1 mm may be a good choice for maintaining the quality of stressed sperm samples, unlike Trolox, which inhibited sperm motility.     

Toward integrated assessment in agriculture

Practitioners of agriculture increasingly find themselves in conflict with the broader society over issues such as siting of livestock rearing facilities and the presence of chemicals in water resources. This is probably a function both of urban development encroaching further into agricultural lands, and of the emerging norm of broader participation in environmental decisions. Numerous public debates over these issues shape the ways by which society reconciles its need for agriculture with the inevitable impacts on nature. Scientific knowledge plays an important role in these debates, but no more so than do human values. Agricultural scientists must seek ways to best inform these debates, while respecting the roles of values. The emerging approach of integrated assessment, now in use in larger-scale issues such as global change and acid rain, appears to have applicability to agriculture. A central component of integrated assessment is the integrated assessment model (IAM), a system model incorporating diverse facets of a problem, so interactions among these facets can be seen. The systems tradition in agriculture, including hard and soft variants, may be viewed as precursors to the IAM approach. Academic agriculturists should begin to create the IAMs that are needed to inform the emerging public debates surrounding agriculture, society, and nature.     

Carbon dioxide emission from european estuaries

The partial pressure of carbon dioxide (pCO2) in surface waters and related atmospheric exchanges were measured in nine European estuaries. Averaged fluxes over the entire estuaries are usually in the range of 0.1 to 0.5 mole of CO2 per square meter per day. For wide estuaries, net daily fluxes to the atmosphere amount to several hundred tons of carbon (up to 790 tons of carbon per day in the Scheldt estuary). European estuaries emit between 30 and 60 million tons of carbon per year to the atmosphere, representing 5 to 10% of present anthropogenic CO2 emissions for Western Europe.