A Quantitative Polymerase Chain Reaction Assay for the Detection and Quantification of Epizootic Epitheliotropic Disease Virus (EEDV; Salmonid Herpesvirus 3)

Epizootic epitheliotropic disease virus (EEDV; salmonid herpesvirus [SalHV3]; family Alloherpesviridae) causes a systemic disease of juvenile and yearling Lake Trout Salvelinus namaycush. No cell lines are currently available for the culture and propagation of EEDV, so primary diagnosis is limited to PCR and electron microscopy. To better understand the pervasiveness of EEDV (carrier or latent state of infection) in domesticated and wild Lake Trout populations, we developed a sensitive TaqMan quantitative PCR (qPCR) assay to detect the presence of the EEDV terminase gene in Lake Trout tissues. This assay was able to detect a linear standard curve over nine logs of plasmid dilution and was sensitive enough to detect single-digit copies of EEDV. The efficiency of the PCR assay was 99.4 ± 0.06% (mean ± SD), with a 95% confidence limit of 0.0296 (R² = 0.994). Methods were successfully applied to collect preliminary data from a number of species and water bodies in the states of Pennsylvania, New York, and Vermont, indicating that EEDV is more common in wild fish than previously known. In addition, through the development of this qPCR assay, we detected EEDV in a new salmonid species, the Cisco Coregonus artedi. The qPCR assay was unexpectedly able to detect two additional herpesviruses, the Atlantic Salmon papillomatosis virus (ASPV; SalHV4) and the Namaycush herpesvirus (NamHV; SalHV5), which both share high sequence identity with the EEDV terminase gene. With these unexpected findings, we subsequently designed three primer sets to confirm initial TaqMan qPCR assay positives and to differentiate among EEDV, ASPV, and NamHV by detecting the glycoprotein genes via SYBR Green qPCR.

Received April 20, 2015; accepted November 10, 2015     

Prevalence of antimicrobial resistance among Salmonella on midwest and northeast USA dairy farms

The objective of this study was to determine the prevalence of antimicrobial resistance among Salmonella isolated from dairy herds in New York, Minnesota, Michigan, and Wisconsin, USA. Serogroup and antimicrobial susceptibility characteristics were determined for Salmonella from cattle and environmental samples collected during August 2000–October 2001 as part of a longitudinal study where 129 herds were visited at 2-month intervals. Salmonella isolates were tested (using a broth microdilution method) for susceptibility to amoxicillin/clavulanic acid, ampicillin, ceftiofur, ceftriaxone, cephalothin, chloramphenicol, ciprofloxacin, gentamicin, kanamycin, nalidixic acid, streptomycin, sulfamethoxazole, tetracycline, and trimethoprim/sulfamethoxazole. Of the 1506 isolates tested for minimum inhibitory concentrations to these 14 antimicrobial agents, 81.2% were pan-susceptible and for most herds (81.6%) the predominant antimicrobial resistance pattern was pan-susceptible. At least 1 Salmonella isolate resistant to 5 or more antimicrobial agents was found on 23.6% of herds. This resistance phenotype was most common among serogroups B and E1 and among samples from calves and farmer-designated sick cows. Resistant samples most frequently exhibited resistance to tetracycline, streptomycin, and/or ampicillin. No samples were resistant to ceftriaxone (though 13 were in the intermediate range), and very few samples were resistant to ciprofloxacin (n = 1), nalidixic acid (n = 5), or trimethoprim/sulfamethoxazole (n = 7).     

A Greener UV and Peroxide-Based Chemical Oxygen Demand Test

Water quality assessment typically includes the determination of chemical oxygen demand (COD) by oxidation of organic matter with Cr(VI) in an acidic medium followed by digestion. Unfortunately, the required reagents are harmful and the reaction times are rather long. We investigated earlier the use of H₂O₂ as a more environmentally friendly oxidizing agent to replace the hazardous chromates. In the present study, we have furthered this possibility by incorporating the use of H₂O₂ in the presence of UV light. A protocol has been devised and tested with standards and real samples that replaces toxic Cr(VI), halves the amount of silver sulfate required, and greatly reduces the necessary reaction time, thus yielding a faster and more environmentally sound method.     

Multi-model comparison on the effects of climate change on tree species in the eastern U.S.: results from an enhanced niche model and process-based ecosystem and landscape models

Context

Species distribution models (SDM) establish statistical relationships between the current distribution of species and key attributes whereas process-based models simulate ecosystem and tree species dynamics based on representations of physical and biological processes. TreeAtlas, which uses DISTRIB SDM, and Linkages and LANDIS PRO, process-based ecosystem and landscape models, respectively, were used concurrently on four regional climate change assessments in the eastern Unites States.

Objectives

We compared predictions for 30 species from TreeAtlas, Linkages, and LANDIS PRO, using two climate change scenarios on four regions, to derive a more robust assessment of species change in response to climate change.

Methods

We calculated the ratio of future importance or biomass to current for each species, then compared agreement among models by species, region, and climate scenario using change classes, an ordinal agreement score, spearman rank correlations, and model averaged change ratios.

Results

Comparisons indicated high agreement for many species, especially northern species modeled to lose habitat. TreeAtlas and Linkages agreed the most but each also agreed with many species outputs from LANDIS PRO, particularly when succession within LANDIS PRO was simulated to 2300. A geographic analysis showed that a simple difference (in latitude degrees) of the weighted mean center of a species distribution versus the geographic center of the region of interest provides an initial estimate for the species’ potential to gain, lose, or remain stable under climate change.

Conclusions

This analysis of multiple models provides a useful approach to compare among disparate models and a more consistent interpretation of the future for use in vulnerability assessments and adaptation planning.

     

Effects of Pre-Stratification Storage Conditions on Black Walnut Seed Post-Stratification Germination Capacity

The objective of this work was to quantify changes in the germination capacity of Juglans nigra seeds following storage and then artificial stratification. Seeds were stored at 5 and –20 °C under 5% and 14% internal moisture content (IMC) for 0, 8, 10, and 12 months under conditions with oxygen or under vacuum. Germination percentage (GP) was affected by storage temperature, IMC, and storage period. Seeds stored with 5% IMC succeeded in germinating regardless of the temperature. Seeds stored with 14% IMC did not germinate (–20 °C) or germinated during storage (5 °C). The GP was reduced with the time of storage in seeds at –20 °C and 5% IMC and this effect was avoided in storage under vacuum. In conclusion, the germination capacity of black walnut seeds did not change for up to 12 months at low above-zero temperature and low IMC (5 °C, 5% IMC), being the most cost-effective alternative for long-term storage of black walnut seeds in commercial nurseries. Vacuum storage avoided the decrease of around 50% in average germination capacity of seeds stored for a year at –20 °C.