Detection and quantification of equine herpesvirus-1 viremia and nasal shedding by real-time polymerase chain reaction.

Equine herpesvirus-1 (EHV-1) infection is common in young horses throughout the world, resulting in respiratory disease, epidemic abortion, sporadic myelitis, or latent infections. To improve on conventional diagnostic tests for EHV-1, a real-time polymerase chain reaction (PCR) technique was developed, using primers and probes specific for the EHV-1 gB gene. Amplification efficiencies of 100% +/- 5% were obtained for DNA isolated from a plasmid, infected peripheral blood mononuclear cells (PBMCs), and nasal secretions from infected ponies. The dynamic range of the assay was 8 log10 dilutions, and the lower limit of detection was 6 DNA copies. Fifteen ponies, seronegative for EHV-1, were experimentally infected with EHV-1, and nasal samples were used to quantify shedding of virus by both virus isolation and real-time PCR analysis. Virus isolation identified nasal shedding of EHV-1 in 12/15 ponies on a total of 25 days; real-time PCR detected viral shedding in 15/15 ponies on 75 days. Viremia was quantified using PBMC DNA, subsequent to challenge infection in 3 additional ponies. Viremia was identified in 1/3 ponies on a single day by virus isolation; real-time PCR detected viremia in 3/3 ponies on 17 days. When real-time PCR was used to analyze PBMC DNA from 11 latently infected ponies (documented by nested PCR), EHV-1 was not detected. We conclude that real-time PCR is a sensitive and quantitative test for EHV-1 nasal shedding and viremia and provides a valuable tool for EHV-1 surveillance, diagnosis of clinical disease, and investigation of vaccine efficacy.     

Performance of a foot-and-mouth disease virus reverse transcription-polymerase chain reaction with amplification controls between three real-time instruments.

The foot-and-mouth disease virus (FMDV) is a member of the picornavirus family, possessing an 8-kb single-stranded RNA genome of positive polarity. It is highly contagious among several livestock species and can lead to severe economic consequences, as evidenced by the UK outbreak in 2001. The usage of real-time polymerase chain reaction has facilitated rapid detection of FMDV. Several real-time PCR instruments are available with various capabilities, such as portability and high sample volume analysis. Primers and a dual-labeled TaqMan probe were optimized to detect a single, highly conserved 88-bp segment of the FMDV 3D (RNA polymerase) gene. To increase the confidence of the RT-PCR result, a positive amplification control was synthesized to detect potential false-positive results due to contamination if a wild-type virus is used as positive control. In addition, a preventative measure against false-negative results was developed in which endogenous beta actin mRNA is coamplified by RT-PCR. Assay performance was compared on the LightCycler1.2 (Roche), the SmartCyclerII (Cepheid), and the SDS 7900HT (ABI). These assays successfully identified the FMDV genome and beta actin mRNA from several sources of infected nasal and oral swabs, as well as probang samples.     

Diagnosis and treatment of presumptive postobstructive pulmonary edema in a Florida panther (Puma concolor coryi).

A free-ranging, adult male Florida panther (Puma concolor coryi) was immobilized and evaluated for hematuria following routine capture. Prior to anesthetic recovery, the panther was fitted with a telemetry collar. After an initially quiet recovery, the panther began thrashing in the transport cage, and was again immobilized. Pink foam was evident from the nostrils, and crackles were ausculted over the chest, indicating pulmonary edema. Postobstructive pulmonary edema was diagnosed based on history, clinical signs, radiographic evaluation, and blood gas analysis. The animal was treated intensively for several hours with diuretics, oxygen, and manual ventilation. The panther responded rapidly to therapy and was released back into the wild 48 hr after presentation. Postobstructive pulmonary edema, also called negative-pressure pulmonary edema, may be underrecognized in veterinary medicine. In this case, the telemetry collar, in conjunction with anesthetic recovery in a small transport crate, may have contributed to tracheal obstruction. Wildlife veterinarians and biologists should be aware of the risk of airway obstruction when placing tracking collars, and animals should be continuously monitored during anesthetic recovery to ensure the presence of a patent airway.     

Disease Assessment Concepts and the Advancements Made in Improving the Accuracy and Precision of Plant Disease Data

New concepts in phytopathometry continue to emerge, such as the evolution of the concept of pathogen intensity versus the well-established concept of disease intensity. The concept of pathogen severity, defined as the quantitative measurement of the amount of pathogen per sampling unit has also emerged in response to the now commonplace development of quantitative molecular detection tools. Although the concept of disease severity, i.e., the amount of disease per sampling unit, is a well-established concept, the accuracy and precision of visual estimates of disease severity is often questioned. This article will review disease assessment concepts, as well as the methods and assessment aides currently available to improve the accuracy and precision of visually-based disease severity data. The accuracy and precision of visual disease severity assessments can be improved by quantitatively measuring and comparing the accuracy and precision of rates and/or assessment methods using linear regression, by using computer-based disease assessment training programmes, and by developing and using diagrammatic keys (standard area diagrams).     

Landscape patterns and agriculture: modelling the long-term effects of human practices on Pinus sylvestris spatial dynamics (Causse Mejean, France)

This paper focuses on understanding human impact on landscape. Both ecological and human practices are analysed as interacting processes. An agent-based model integrating biological and historical knowledge is used to analyse the pattern of Scots Pine encroachment in a French Mediterranean upland. In the STIPA model, pine trees are autonomous agents and a cellular automaton simulates land-use. We test the effects of shifting cultivation on tree establishment at the landscape scale. This allows us to understand how agropastoral practices patterned this area from the 17th to 19th century: simulations show the importance of shifting cultivation in limiting woodland progression. Fallow duration linked to environmental heterogeneity is a significant factor for explaining pine dynamics and landscape patterning at the scale of the study region. We put this result in perspective with current rangeland management policies that often consider grazing as the most relevant tool for open landscape maintenance. Our results also show the importance of taking into account time-scale effects when linking landscape patterns to agricultural systems.     

Analysis and modeling of gas exchange processes in Scaevola aemula

Scaevola aemula is a popular ornamental crop cultivated as a bedding plant or for hanging baskets. We characterized gas exchange properties of S. aemula ‘New Wonder’ in response to photosynthetically active radiation (PAR), carbon dioxide concentration, and leaf temperature. Net CO₂ assimilation rate (A) was responsive to CO₂, exhibiting a saturation when intercellular CO₂ concentration (C_i) was greater than 600 μmol mol⁻¹. Net CO₂ assimilation rate and dark respiration rate (R_d) were 23.1 and 2.3 μmol m⁻² s⁻¹, respectively, at 25 °C and PAR = 1500 μmol m⁻² s⁻¹. Net CO₂ assimilation rates were similar at leaf temperatures between 20 and 30 °C but significantly reduced at 15 °C. These gas exchange results were used to test the extendibility of a coupled gas exchange model previously developed for cut-roses. Utilizing the gas exchange data measured at 25 °C leaf temperature, several model parameters were independently determined for S. aemula. Model predictions were then compared with observations at different leaf temperatures. The model predicted the rates of net CO₂ assimilation and transpiration of S. aemula reasonably well. Without additional calibration, the model was capable of predicting the temperature dependence of net CO₂ assimilation and transpiration rates. Applying the model to predict the effects of supplemental lighting and CO₂ enrichment on canopy photosynthesis and transpiration rates, we show that this model could be a useful tool for examining environmental control options for S. aemula production in the greenhouse.     

Empirical relationships between land use/cover and estuarine condition in the Northeastern United States

Land–water interactions were examined in three regions in the Virginian Biogeographic Province; the southern shore of Cape Cod, Massachusetts; the Hudson/Raritan region of New York; and the eastern shore of the Delmarva (Delaware/Maryland/Virginia) Peninsula. Cumulative distribution functions were used to evaluate similarity in environmental condition among estuaries. Spatial-setting variables (location in a river, coastal lagoon, or in open waters) were associated with variation for some measures of estuarine condition. Patterns of coastal urban and agriculture gradients were measured and their relationship with indicators of estuarine condition was modeled statistically. When estuaries were pooled, the highest variation explained by spatial-setting variables was found for dissolved oxygen (DO, R ² = 0.44) and salinity (R ² = 0.58), with DO decreasing in river locations and salinity decreasing with rainfall and sampling locations near rivers. The explanatory power for the other indicator variables was low and varied from 6% to 27%. Rainfall explained some of the variation (R ² = 0.23) in total suspended solids. Moderate (0.4 < | r | < 0.7) to strong (| r | ≥ 0.7) linear associations were found between total urban area and measures of estuarine condition. Within regions, total urban area was positively associated with Silver (r = 0.59), Cadmium (r = 0.65), and Mercury (r = 0.47) in Cape Cod, and inversely related to DO (r = −0.65) in the Hudson/Raritan region. No associations were found in the Delmarva Peninsula study area. Total area of agriculture showed a moderate association with Arsenic in Cape Cod, but no other associations were found in the other two regions. Our analyses show a measurable impact of urban land use on coastal ecosystem condition over large areas of the northeastern United States. This pattern was most evident when many different landscapes were considered simultaneously. The relationship between urban development and estuarine condition were weaker within the individual regions studied. The use of land use/cover models for predicting estuarine condition is a challenging task that warrants enhancements in the type, quantity, and quality of data to improve our ability to discern relationships between anthropogenic activities on land and the condition of coastal environments.