Pen-Based Swine Oral Fluid Samples Contain Both Environmental and Pig-Derived Targets.

Laboratory methods for detecting specific pathogens in oral fluids are widely reported, but there is little research on the oral fluid sampling process itself. In this study, a fluorescent tracer (diluted red food coloring) was used to test the transfer of a target directly from pigs or indirectly from the environment to pen-based oral fluid samples. Pens of ~30, ~60, and ~125 14-week-old pigs (32 pens/size) on commercial swine farms received one of two treatments: (1) pig exposure, i.e., ~3.5 mL of tracer solution sprayed into the mouth of 10% of the pigs in the pen; (2) environmental exposure, i.e., 20 mL of tracer solution was poured on the floor in the center of the pen. Oral fluids collected one day prior to treatment (baseline fluorescence control) and immediately after treatment were tested for fluorescence. Data were evaluated by receiver operating characteristic (ROC) analysis, with Youden's J statistic used to set a threshold. Pretreatment oral fluid samples with fluorescence responses above the ROC threshold were removed from further analysis (7 of 96 samples). Based on the ROC analyses, oral fluid samples from 78 of 89 pens (87.6%), contained red food coloring, including 43 of 47 (91.5%) pens receiving pig exposure and 35 of 42 (83.3%) pens receiving environmental exposure. Thus, oral fluid samples contain both pig-derived and environmental targets. This methodology provides a safe and quantifiable method to evaluate oral fluid sampling vis-à-vis pen behavior, pen size, sampling protocol, and target distribution in the pen.

Static Aerated Composting of African Swine Fever Virus-Infected Swine Carcasses with Rice Hulls and Sawdust.

Identifying and ensuring the inactivation of the African Swine Fever virus in deadstock is a gap in the swine industry's knowledge and response capabilities. The results of our study demonstrate that ASFv in deadstock was inactivated using static aerated composting as the carcass disposal method. Replicated compost piles with whole market hogs and two different carbon sources were constructed. In-situ bags containing ASFv-infected spleen tissue were placed alongside each of the carcasses and throughout the pile. The bags were extracted at days 0, 1, 3, 7, 14, 28, 56, and 144 for ASFv detection and isolation. Real-time PCR results showed that DNA of ASFv was detected in all samples tested on day 28. The virus concentration identified through virus isolation was found to be below the detection limit by day 3 in rice hulls and by day 7 in sawdust. Given the slope of the decay, near-zero concentration with 99.9% confidence occurred at 5.0 days in rice hulls and at 6.4 days in sawdust. Additionally, the result of virus isolation also showed that the virus in bone marrow samples collected at 28 days was inactivated.

Viability of African Swine Fever Virus with the Shallow Burial with Carbon Carcass Disposal Method.

African swine fever (ASF) is a highly contagious swine disease with high mortality. In many countries, culling pigs infected and exposed to the ASF virus is mandatory to control the disease, which poses a real challenge in the disposal of large numbers of carcasses during ASF outbreaks. Shallow burial with carbon (SBC) Thanks ew mortality disposal method developed from deep burial and composting. The present study investigates the effectiveness of SBC in disposing of ASF virus-infected pigs. The real-time PCR results showed that DNA of the ASF virus was still detected in bone marrow samples on day 56, while the virus isolation test revealed that the infectious ASF virus was destroyed in both spleen and bone marrow samples on day 5. Interestingly, decomposition was found to occur rapidly in these shallow burial pits. On day 144, only large bones were found in the burial pit. In general, the results of this study indicated that SBC is a potential method for the disposal of ASF-infected carcasses; however, further studies are needed to provide more scientific evidence for the efficacy of SBC in different environment conditions.

Virus viability in spiked swine bone marrow tissue during above-ground burial method and under in vitro conditions.

The emergence of high consequence animal diseases usually requires managing significant mortality. A desirable aspect of any carcass management method is the ability to contain and inactivate the target pathogen. The above-ground burial (AGB) technique was recently developed and proposed as an alternative carcass management method. Here, we investigate the tenacity of swinepox virus (SwPV), as a surrogate model for African swine fever virus (ASFV) in swine carcasses during the AGB process. For this, SwPV was inoculated intrafemorally in 90 adult swine carcasses, which were subsequently disposed under AGB conditions. Bone marrow samples were recovered periodically throughout 12 months and virus viability was assessed by virus isolation (VI), whereas the presence of SwPV DNA was evaluated by quantitative polymerase chain reaction (qPCR). Additionally, an in vitro study assessed the inactivation rate of SwPV, Senecavirus A (SVA), and bovine viral diarrhoea virus (BVDV). Viral suspensions were mixed with bone marrow material and maintained at 21-23°C for 30 days. Virus viability was assessed by VI and viral titration. In the field study, SwPV remained viable only in 11 (55%) bone marrow samples collected on day 7; only viral DNA (and not infectivity) was detected afterwards. SwPV inactivation was estimated to have occurred by day 11. The in vitro testing revealed a variable tenacity of the studied viruses. The viability period was estimated in 28, 80, and 118 days, respectively, for BVDV, SwPV, and SVA. Overall, these findings indicate that the AGB technique was effective in quickly inactivating SwPV. Additionally, the SwPV inactivation rate is comparable to ASFV under field studies and poses a potential model for preliminary ASFV inactivation studies with reduced biosecurity requirements. Moreover, this study contributes to understanding the inactivation kinetics of viruses under specific conditions, which is critical when designing and applying countermeasures in case of biosecurity breaches in sites managing animal mortality.

Assessing the value of PCR assays in oral fluid samples for detecting African swine fever, classical swine fever, and foot-and-mouth disease in U.S. swine.

INTRODUCTION: Oral fluid sampling and testing offers a convenient, unobtrusive mechanism for evaluating the health status of swine, especially grower and finisher swine. This assessment evaluates the potential testing of oral fluid samples with real-time reverse-transcriptase polymerase chain reaction (rRT-PCR) to detect African swine fever, classical swine fever, or foot-and-mouth disease for surveillance during a disease outbreak and early detection in a disease-free setting. METHODS: We used a series of logical arguments, informed assumptions, and a range of parameter values from literature and industry practices to examine the cost and value of information provided by oral fluid sampling and rRT-PCR testing for the swine foreign animal disease surveillance objectives outlined above. RESULTS: Based on the evaluation, oral fluid testing demonstrated value for both settings evaluated. The greatest value was in an outbreak scenario, where using oral fluids would minimize disruption of animal and farm activities, reduce sample sizes by 23%-40%, and decrease resource requirements relative to current individual animal sampling plans. For an early detection system, sampling every 3 days met the designed prevalence detection threshold with 0.95 probability, but was quite costly. LIMITATIONS: Implementation of oral fluid testing for African swine fever, classical swine fever, or foot-and-mouth disease surveillance is not yet possible due to several limitations and information gaps. The gaps include validation of PCR diagnostic protocols and kits for African swine fever, classical swine fever, or foot-and-mouth disease on swine oral fluid samples; minimal information on test performance in a field setting; detection windows with low virulence strains of some foreign animal disease viruses; and the need for confirmatory testing protocol development.

Use of Temperature, Humidity, and Slaughter Condemnation Data to Predict Increases in Transport Losses in Three Classes of Swine and Resulting Foregone Revenue.

The United States Department of Agriculture (USDA) Animal and Plant Health Inspection Service (APHIS) conducts weekly surveillance of slaughter condemnation rates to provide early warning for emerging diseases and to monitor health trends in swine. Swine deaths in-transit are an animal welfare concern and represent lost revenue for the swine industry. This retrospective observational study used ambient temperature and humidity data from weather stations near United States slaughter plants collected from 2010 to 2015 to predict the incidence and risk of death among swine in-transit and just prior to slaughter. The risk of death for market swine at a heat index (HI), which combines the effects of temperature and humidity, indicating moderately hot weather conditions between 85 and 92°F was 1.37 times greater than that of the baseline temperature range of 54-79°F. The risk of death for cull sows at an HI between 85 and 92°F was 1.93 times greater than that of average temperatures ranging from 54 to 79°F. Roaster swine (weigh < 220 lbs and often used for whole carcass roasting), however, had 0.80 times the risk when the HI was 85-92°F compared to a baseline temperature of 54-79°F. The risk of death for roaster swine at a minimum temperature between 40 and 50°F was 1.21 times greater than that of average temperatures ranging from 54 to 79°F. The risk of death for market swine at a minimum temperature range of 40-50°F was 0.97 times that of average temperatures ranging from 54 to 79°F. And for cull sows, the risk of death at a minimum temperature range of 40-50°F was 0.81 times the risk at the average temperature ranging from 54 to 79°F. Across the study period, cumulative foregone revenue, or revenue not realized due to swine condemnations, for all swine was $18.6 million and $4.3 million for cold temperatures and high HI ranges above the baseline, respectively. Marginal foregone revenue per hog in hotter months is higher due to seasonal peaks in swine prices. As a result of this study, the USDA-APHIS swine condemnation surveillance can incorporate weekly estimated HI values and ambient temperature data for slaughter establishments to provide additional information for analysts investigating signals (noteworthy increases above baseline) for "dead" condemnations. This study suggests that current mitigation measures are often not sufficient to prevent swine deaths due to ambient temperature extremes.

Analysis of the diagnostic accuracy of the gamma interferon assay for detection of bovine tuberculosis in U.S. herds.

The goal of this study was to evaluate the test sensitivity (SE) and specificity (SP) of the gamma interferon (G-IFN) assay used for the detection of bovine tuberculosis (bTB) in U.S. cattle herds. In addition, the study assessed the association between G-IFN test results and bTB status of cattle, and explored different cut off values for classification of test results in adult cattle using receiver operating characteristics (ROC) curve analysis. Test SE was estimated using a population of 87 confirmed infected cattle from 14 herds distributed in 6 states. Test SP was estimated using a population of 4123 cattle representing 3000 premises in 3 states. These animals were from bTB free areas, accredited bTB free herds, or herds that were historically bTB free based on the absence of lesions found at slaughter and historical records of negative tests performed for bTB surveillance. The distribution of G-IFN results and its association with bTB infection status was also explored in a group of 914 exposed cattle in which infection was not confirmed. The results showed that the SE of the G-IFN for a cut-off value ≥0.1 was 83.9% (76.1, 91.6). The SP of the G-IFN was 90.7% (95% CI: 89.8, 91.6), 97% (95% CI: 96.5, 97.5), and 98.6%(95% CI: 98.2, 98.9), for cut off values of 0.1, 0.3, and 0.5, respectively. For a cut off value ≥0.1, the likelihood ratio of a positive G-IFN test was 9.03 (95% CI: 7.90, 10.31), and the likelihood ratio of a negative G-IFN test was 0.18 (95% CI: 0.11, 0.29). The area under the ROC curve was 0.976 (95% CI: 0.97, 0.98), characteristic of a highly accurate test. ROC analysis also showed that lower cut-off values, such as 0.1, have high SE with suitable SP for use in parallel testing, while cut-off values ranging between 0.3 and 0.6 provide the high SP desired in series-testing protocols with lower SE values. Findings from this study indicated that the G-IFN performs with high accuracy in the field, yielding SE and SP estimates comparable to those reported in previous evaluations (Ryan et al., 2000; Ameni et al., 2000; de la Rua-Domenech et al., 2006; Gormley et al., 2006).

Plant uptake of depleted uranium from manure-amended and citrate treated soil.

Six plant species were tested for their ability to accumulate depleted uranium in their above-ground biomass from deployed munitions contaminated soil in New Mexico. In greenhouse experiments, Kochia (Kochia scoparia L. Schrad.) and pigweed (Amaranthus retroflexus L) were grown with steer manure added at rates of 22.4, 44.8, and 89.6 Mg ha(-1). Citric acid and glyphosate (N-(phosphonomethyl) glycine) applied at the end of the growing season increased DU concentrations from 2.5 to 17 times. Leaf and stem DU concentrations in kochia increased from 17.0 to 41.9 mg kg(-1) and from 3.5 to 18.0 mg kg(-1), respectively. In pigweed, leaf and stem DU concentrations increased from 1.0 to 17.3 and from 1.0 to 4.7 mg kg(-1), respectively. Manure generally decreased or had no effect on DU uptake. The effect of citric acid and ammonium citrate on DU uptake by kochia, sunflower (Helianthus annuus L), and sweet corn (Zea mays L) was also studied. Ammonium citrate was just as effective in enhancing DU uptake as citric acid. This implies that the citrate ion is more important in DU uptake and translocation than the solubilization of DU through acidification. In both experiments, leaves had higher DU concentrations than stems.