Optimization of a Plaque Neutralization Test (PNT) to Identify the Exposure History of Pacific Herring to Viral Hemorrhagic Septicemia Virus (VHSV)

Abstract

Methods for a plaque neutralization test (PNT) were optimized for the detection and quantification of viral hemorrhagic septicemia virus (VHSV) neutralizing activity in the plasma of Pacific Herring Clupea pallasii. The PNT was complement dependent, as neutralizing activity was attenuated by heat inactivation; further, neutralizing activity was mostly restored by the addition of exogenous complement from specific-pathogen-free Pacific Herring. Optimal methods included the overnight incubation of VHSV aliquots in serial dilutions (starting at 1:16) of whole test plasma containing endogenous complement. The resulting viral titers were then enumerated using a viral plaque assay in 96-well microplates. Serum neutralizing activity was virus-specific as plasma from viral hemorrhagic septicemia (VHS) survivors demonstrated only negligible reactivity to infectious hematopoietic necrosis virus, a closely related rhabdovirus. Among Pacific Herring that survived VHSV exposure, neutralizing activity was detected in the plasma as early as 37 d postexposure and peaked at approximately 64 d postexposure. The onset of neutralizing activity was slightly delayed in fish reared at 7.4°C relative to those in warmer temperatures (9.9°C and 13.1°C); however, neutralizing activity persisted for at least 345 d postexposure in all temperature treatments. It is anticipated that this novel ability to assess VHSV neutralizing activity in Pacific Herring will enable retrospective comparisons between prior VHS infections and year-class recruitment failures. Additionally, the optimized PNT could be employed as a forecasting tool capable of identifying the potential for future VHS epizootics in wild Pacific Herring populations.

Received November 7, 2016; accepted January 14, 2017 Published online April 4, 2017     

Effects of sensor spatial resolution on landscape structure parameters

We examined the effects of increasing grain size from 20 m to 1100 m on landscape parameters characterizing spatial structure in the northern Wisconsin lake district. We examined whether structural parameters remain relatively constant over this range and whether aggregation algorithms permit extrapolation within this range. Images from three different satellite sensors were employed in this study: (1) the SPOT multispectral high resolution visible (HRV), (2) the Landsat Thematic Mapper (TM), and (3) the NOAA Advanced Very High Resolution Radiometer (AVHRR). Each scene was classified as patches of water in a matrix of land. Spatial structure was quantified using several landscape parameters: percent water, number of lakes (patches), average lake area and perimeter, fractal dimension, and three measures of texture (homogeneity, contrast, and entropy). Results indicate that most measures were sensitive to changes in grain size. As grain size increased from 20 m using HRV image data to 1100 m (AVHRR), the percent water and the number of lakes decreased while the average lake area, perimeter, the fractal dimension, and contrast increased. The other two texture measures were relatively invariant with grain size. Although examination of texture at various angles of adjacency was performed to investigate features which vary systematically with angle, the angle did not have an important effect on the texture parameter values. An aggregation algorithm was used to simulate additional grain sizes. Grain was increased successively by a factor of two from 20 m (the HRV image) to 1280 m. We then calculated landscape parameter values at each grain size. Extrapolated values closely approximated the actual sensor values. Because the grain size has an important effect on most landscape parameters, the choice of satellite sensor must be appropriate for the research question asked. Interpolation between the grain sizes of different satellite sensors is possible with an approach involving aggregation of pixels.     

The Effects of Sample Size on the Outcome of Ovarian Tissue Cryopreservation

Cryopreservation of ovarian tissue is known to affect follicular survival. Several variables may be responsible for this. Little attention has focused on the effect of the size of the fragment to be cryopreserved. This study was conducted to assess the effect of the size of the tissue on follicular histology after freezing with 1,2-propanediol. Histological evaluations were performed of control and cryopreserved tissue. Fragments were cut 10 × 3 × 2 mm³ (2 mm group) or 10 × 3 × 4 mm³ (4 mm group). Percentages of normal follicles in control fragments cut into 2 and 4 mm slices were 56% and 34%, respectively. The relative risks to obtain normal follicles in the 2 mm and the 4 mm fragments after cryopreservation were 0.63 and 0.47, respectively. Freezing reduced follicle survival to a significantly greater extent in the larger tissue fragments. There is an increased risk of damage to primary and primordial follicles, when the tissue slices are cut with all dimensions larger than 2 mm.     

Management-induced change in labile soil organic matter under continuous corn in eastern Canadian soils

Soil samples taken from four experimental sites that had been cropped to continuous corn for 3–11 years in Ontario and Quebec were analyzed to evaluate changes in quantity and quality of labile soil organic carbon under different nitrogen (N) fertility and tillage treatments. Addition of fertilizer N above soil test recommendations tended to decrease amounts of water-soluble organic carbon (WSOC) and microbial biomass carbon (MBC). The quality of the WSOC was characterized by ¹³C nuclear magnetic resonance and infrared spectrophotometry and the results indicated that carbohydrates, long-chain aliphatics and proteins were the major components of all extracts. Similar types of C were present in all of the soils, but an influence of management was evident. The quantity of soil MBC was positively related to the quantities of WSOC, carbohydrate C, and organic C, and negatively related to quantities of long-chain aliphatic C in the soil. The quantity of WSOC was positively related to the quantities of protein C, carbohydrate C, and negatively related to the quantity of carboxylic C. The quantity of soil MBC was not only related to quantities of soil WSOC but also to the quality of soil WSOC. Received: 2 April 1997     

Environmental conditions and isozyme polymorphism in Chenopodium album L.

Electrophoretic variations within four enzyme systems (α-esterase, acid phosphatase, malate dehydrogenase and leucine amino peptidase), were studied for various garden and field populations of Chenopodium album. A correlation was found between the degree of polymorphism for the electrophoretic traits and environmental conditions. The amount of phenotypic polymorphism is very high in gardens, intermediate in an untreated and, low in herbicide-treated fields and nil for a triazine resistant population in a maize (Zea mats L.) monoculture. Furthermore, the tetraploid versus hexaploid ratio is higher within garden populations. Higher variability in garden populations is due, in part, to a high proportion of polymorphic phenotypes, and in part to a low effect of selection pressure in such types of habitat. Although there is a difference between phenotypes of populations from acid soils and those from more basic ones, we cannot be sure that the variation of polymorphism is due only to selection.     

Developing stochastic epidemiological models to quantify the dynamics of infectious diseases in domestic livestock

A stochastic model describing disease transmission dynamics for a microparasitic infection in a structured domestic animal population is developed and applied to hypothetical epidemics on a pig farm. Rational decision making regarding appropriate control strategies for infectious diseases in domestic livestock requires an understanding of the disease dynamics and risk profiles for different groups of animals. This is best achieved by means of stochastic epidemic models. Methodologies are presented for 1) estimating the probability of an epidemic, given the presence of an infected animal, whether this epidemic is major (requires intervention) or minor (dies out without intervention), and how the location of the infected animal on the farm influences the epidemic probabilities; 2) estimating the basic reproductive ratio, R0 (i.e., the expected number of secondary cases on the introduction of a single infected animal) and the variability of the estimate of this parameter; and 3) estimating the total proportion of animals infected during an epidemic and the total proportion infected at any point in time. The model can be used for assessing impact of altering farm structure on disease dynamics, as well as disease control strategies, including altering farm structure, vaccination, culling, and genetic selection.     

Stability of resistance to Phytophthora infestans in potato: an international evaluation

Ten institutions in nine countries joined together to test the stability of resistance of 14 potato genotypes to the oomycete pathogen Phytophthora infestans in three separate trials. Seven of the genotypes were tested in one trial involving seven locations, and all 14 were tested in two subsequent trials, each involving eight locations. Stability of resistance was tested with nonparametric tests and with an additive main effects and multiplicative interaction (AMMI) model. Overall, resistance to P. infestans was robust; resistant genotypes were consistently resistant in all locations and trials. The nonparametric analysis indicated that specific genotypes were basically stable across sites for resistance. In trial 3, the Z statistic for overall stability was significant at 0·05%, indicating a significant level of interaction across the trial, but there were no significant interactions for specific genotypes in this trial. The genotype by environment (G × E) effect of the AMMI model was highly significant in both trials, but the mean square of G × E was less than 10% of the genotype effect in each trial. The first two principal components (PCA1 and PCA2) of the AMMI analyses together explained 75 and 80% of the interaction effects in trials 2 and 3, respectively. Based on both nonparametric and AMMI analyses, Ecuador and Argentina were locations of relatively high interaction effects for both trials 2 and 3, although in Ecuador this interaction was not associated with any particular potato genotype. Other locations also had high interaction effects, but these occurred in only one trial. The genotypes Chata Blanca and, to a lesser extent, Torridon were relatively unstable in trials 2 and 3, but in the case of Torridon, resistant, this did not represent a significant loss of resistance.