Comparative analysis of replication characteristics of BoHV-1 subtypes in bovine respiratory and genital mucosa explants: a phylogenetic enlightenment

ABSTRACT: In general, members of the Alphaherpesvirinae use the epithelium of the upper respiratory and/or genital tract as a preferential site for primary replication. Bovine herpesvirus type 1 (BoHV-1) may replicate at both sites and cause two major clinical entities designated as infectious bovine rhinotracheitis (IBR) and infectious pustular vulvovaginitis/balanoposthitis (IPV/IPB) in cattle. It has been hypothesized that subtype 1.1 invades preferentially the upper respiratory mucosa whereas subtype 1.2 favors replication at the peripheral genital tract. However, some studies are in contrast with this hypothesis. A thorough study of primary replication at both mucosae could elucidate whether or not different BoHV-1 subtypes show differences in mucosa tropism. We established bovine respiratory and genital organ cultures with emphasis on maintenance of tissue morphology and viability during in vitro culture. In a next step, bovine respiratory and genital mucosa explants of the same animals were inoculated with several BoHV-1 subtypes. A quantitative analysis of viral invasion in the mucosa was performed at 0 h, 24 h, 48 h and 72 h post inoculation (pi) by measuring plaque latitude and penetration depth underneath the basement membrane. All BoHV-1 subtypes exhibited a more profound invasion capacity in respiratory tissue compared to that in genital tissue at 24 h pi. However, at 24 h pi plaque latitude was found to be larger in genital tissue compared to respiratory tissue and this for all subtypes. These similar findings among the different subtypes take the edge off the belief of the existence of specific mucosa tropisms of different BoHV-1 subtypes.     

The Use of Decapsulated Cysts of the Brine Shrimp Artemia as Direct Food for Carp Cyprinus carpio L. Larvae

Decapsulated Arremia cysts have been evaluated as a direct food source for larvae of the carp Cyprinus carpio L. All decapsulated cyst diets gave excellent survival of carp larvae during the first two weeks of culturing. Unlike traditionally brine-stored decapsulated cysts, dried Artemia embryos provided growth results comparable to those obtained with freshly hatched Artemia nauplii. Furthermore, except for Artemia embryos that lost their hatchability after long-term storage in air, several other inactivation treatments, simulating improper harvesting and processing of cysts, did not produce a significant decrease in the nutritional quality of the decapsulated cysts.
Using decapsulated cysts 89 a direct food source for carp larvae, instead of nauplii, the quantity of cysts needed can be reduced by about 25% to 35% after one and two weeks of culturing, respectively. Moreover, the use of decapsulated cysts advances the possible commercialition of poor-hatching and less expensive cyst products for culture of carp larvae.     

Limitations of Irrigation Water Quality Guidelines from a Multiple Use Perspective

The standards for irrigation water qualityare currently provided by global andnational guidelines that are mostly basedon the assumption that threshold valuescould be applied to protect crops. Thisapproach can create problems for a largelyunrecognized group of people who make useof irrigation water for non-agriculturalpurposes. At the same time, increasingwater scarcity will lead to the need forrecycling of water in irrigated riverbasins, and the use of low qualityirrigation water. Apart from hazards ofhigh pollutant levels, a sustainabilitycriterion has to be included in the waterquality guidelines to account for long-termlow-level application of certain pollutantsthat can accumulate in the environment.Using the example of cadmium, it is arguedthat the current guidelines need to berevised and should take local factors andfuture developments into account.     

Control and detection of singlet-triplet mixing in a random nuclear field

We observed mixing between two-electron singlet and triplet states in a double quantum dot, caused by interactions with nuclear spins in the host semiconductor. This mixing was suppressed when we applied a small magnetic field or increased the interdot tunnel coupling and thereby the singlet-triplet splitting. Electron transport involving transitions between triplets and singlets in turn polarized the nuclei, resulting in marked bistabilities. We extract from the fluctuating nuclear field a limitation on the time-averaged spin coherence time T2* of 25 nanoseconds. Control of the electron-nuclear interaction will therefore be crucial for the coherent manipulation of individual electron spins.     

Ex-post evaluation of erosion control measures in southern Mali

As part of an impact study of a soil and water conservation (SWC) project in southern Mali, the effect of erosion control measures on soil erosion was evaluated. In one village, a baseline situation from 1988 was compared with the situation in 2003, after farmers had installed stone rows, live fences and grass strips, and had started cultivating across to the slope. This comparison showed a spectacular decrease in gully volume in cultivated fields of 87%, from 58 to 8 m³ ha⁻¹. Estimated annual soil loss decreased with 77% from 42 to 10 t ha⁻¹ year⁻¹. However, baseline data on erosion gullies were not available for other villages. In the absence of baseline data, a simple ‘with–without’ comparison does not allow a correct evaluation because farmers install erosion control measures especially in fields with more erosion. Two alternative methods were used: a reconstructed baseline and a virtual time series. Using the reconstructed baseline, looking not only at active gullies but also at (partly) reclaimed gullies, we concluded that line interventions, gully interventions, and a combination of both, reduced the proportion of active gullies by 48%, 47% and 70%, respectively. Using a virtual time series, comparing erosion in fields with erosion control measures installed in different years, we concluded that erosion gradually decreased by 50% during the 3 years after installation of erosion control measures. In a separate study, we found a positive effect of gully interventions and sowing across to the slope in reducing sheet erosion. A reduction of the slope of the sowing direction by 1% reduced the cover of runoff deposit of coarse sand with 8%. Although a documented baseline is preferred, both a reconstructed baseline and a virtual time series are useful tools and make ex-post evaluations more relevant than a simple with–without comparison.     

Long-term dynamics and population viability in one of the last populations of the endangered Spiranthes spiralis (Orchidaceae) in the Netherlands

During the last decades, most orchid species in much of Western Europe have suffered significant declines and the long-term survival of the remaining populations remains to a large extent uncertain. In particular, populations at range margins may be more prone to extinction than more central populations, as the former tend to be small and isolated, occur in ecologically marginal habitats and have a lower per-capita reproductive rate. In this study, we investigated the long-term dynamics and population viability of a population at the margin of its range of Spiranthes spiralis in the Netherlands. At present, only 2 out of 40 previously known populations persist. Individual plants were monitored for 24 years and their life span, flowering frequency and vegetative growth were determined. Individual plants showed large temporal variation in sexual and vegetative growth among years. The proportion of flowering plants varied from 0 (no plants were flowering) to 100 (all plants were flowering). Vegetative growth, on the other hand, increased when the number of individuals decreased. Dormancy was present, but occurred only in a few individuals. Using a non-structured population viability model, future prospects of this species were assessed. Calculation of extinction probabilities and estimated times to extinction using the diffusion approximation model showed that the species had a relatively high probability (79%) of surviving the next 20 years, whereas the median time to extinction was forty years. However, because 95% confidence intervals of the population growth included 1, we suggest that continued monitoring and additional genetic research are needed to assess the long-term viability of this species.     

A simple and accurate method for determining wheat grain fructan content and average degree of polymerization

An improved method for the measurement of fructans in wheat grains is presented. A mild acid treatment is used for fructan hydrolysis, followed by analysis of the released glucose and fructose with high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD). Not only the amount of fructose set free from fructans but also the released glucose can be quantified accurately, allowing determination of the average degree of polymerization of fructans (DP(av)). Application of the mild acid treatment to different grain samples demonstrated that a correction should be made for the presence of sucrose and raffinose, but not for stachyose or higher raffinose oligosaccharides. The fructan content and DP(av) of spelt flour, wheat flour, and whole wheat flour were 0.6%, 1.2%, and 1.8% of the total weight and 4, 5, and 6, respectively. Validation experiments demonstrate that the proposed quantification method is accurate and repeatable and that also the DP(av) determination is precise.     

Community patterns of soil bacteria and nematodes in relation to geographic distance

Ecosystems consist of aboveground and belowground subsystems and the structure of their communities is known to change with distance. However, most of this knowledge originates from visible, aboveground components, whereas relatively little is known about how soil community structure varies with distance and if this variability depends on the group of organisms considered. In the present study, we analyzed 30 grasslands from three neighboring chalk hill ridges in southern UK to determine the effect of geographic distance (1–198 km) on the similarity of bacterial communities and of nematode communities in the soil. We found that for both groups, community similarity decayed with distance and that this spatial pattern was not related to changes either in plant community composition or soil chemistry. Site history may have contributed to the observed pattern in the case of nematodes, since the distance effect depended on the presence of different nematode taxa at one of the hill ridges. On the other hand, site-related differences in bacterial community composition alone could not explain the spatial turnover, suggesting that other factors, such as biotic gradients and local dispersal processes that we did not include in our analysis, may be involved in the observed pattern. We conclude that, independently of the variety of causal factors that may be involved, the decay in similarity with geographic distance is a characteristic feature of both communities of soil bacteria and nematodes.