DNA Vaccination Partially Protects Muskellunge against Viral Hemorrhagic Septicemia Virus (VHSV-IVb)

Abstract

A DNA vaccine containing the glycoprotein (G) gene of the North American viral hemorrhagic septicemia virus (VHSV) genotype IVb was developed to evaluate the immune response of fish following vaccination and evaluate its efficacy in protecting a susceptible species, the Muskellunge Esox masquinongy, against VHSV-IVb challenge. Seven weeks (539 degree-days) following vaccination with 10 μg of either pVHSivb-G or a control plasmid, Muskellunge were challenged by immersion with 10⁵ plaque-forming units (pfu)/mL of VHSV-IVb. Fish vaccinated with pVHSivb-G had a relative percent survival (RPS) of 45%. Vaccinated fish also had significantly lower mean viral titers in tissues (4.2 × 10² pfu/g) and viral prevalence (4%) than fish receiving the plasmid control vaccine (3.3 × 10⁵ pfu/g; 82%). Neutralizing antibodies were detected 28 d (308 degree-days) postchallenge (11 weeks postvaccination) in 100% of Muskellunge vaccinated with pVHSivb-G compared with only 12% of plasmid-control-vaccinated Muskellunge, suggesting robust induction of a secondary, adaptive immune response. In addition, pVHSivb-G–vaccinated Rainbow Trout Oncorhynchus mykiss challenged 7 d (100 degree-days) postvaccination with the heterologous novirhabdovirus, infectious hematopoietic necrosis virus (IHNV), experienced an RPS of 61%, compared to control fish, suggesting induction of an early and transient nonspecific antiviral immune response. This study provides an important starting point for VHSV-IVb vaccine development and useful information about the antiviral immune response elicited by DNA vaccination in a nondomesticated fish species.

Received May 1, 2016; accepted September 1, 2016     

Livestock and livelihoods of smallholder cattle-owning households in Cambodia: the contribution of on-farm and off-farm activities to income and food security

The majority of smallholder farming households in Cambodia are rurally based and rely on agriculture to support their livelihoods. However, in recent years, growth in the agriculture sector has stagnated with farmers facing several challenges including declining prices for traditional crops and irregular rainfall patterns. This has led to a need for farmers to diversify income sources with livestock promoted as a more viable livelihood activity, particularly the raising of cattle and poultry. However, uncertain profitability of livestock activities is a common perception by smallholders, especially where animals have not been traditionally viewed as a primary income source. To address this, information is required which compares the income and expenses associated with livestock raising to other on-farm activities and off-farm sources. This study reports on a survey of livelihood survey of 17 male and 21 female representatives of 20 households owning cattle in Cambodia, comparing the associated income and expenses of the various livelihood activities in the 12-month period from January to December 2016. Combined total household income from both on-farm and off-farm sources ranged from USD875 to 17730 with an average of USD6779, representing 51% and 49% from on-farm and off-farm activities, respectively. Cattle raising was the most common source of on-farm income (85%), contributing USD1064 and representing 22% of total household income. General household expenses, such as food and transportation (including expenses associated with off-farm employment), represented the majority of total household expenses (79%). Gross profit calculations indicated higher values for pig raising (USD1841.79), cattle (USD950.80) and non-rice crops (USD884) whilst the highest gross margin value was recorded for cattle (89.33%) followed by vegetables (85.27%) and non-rice crops (83.08%). Whilst wet season and dry season rice returned a negative gross profit value of USD197.27 and USD90.60 on average per household, respectively, both were ranked as most important for household consumption, followed by poultry, providing the main source of energy (rice) and protein (chicken meat) and sustaining household food requirements. The study concludes that although smallholder cattle-owning households in Cambodia undertake a diverse range of on-farm activities, as cattle raising provides a superior income source due to higher returns and lower variable costs, it could be promoted as a preferred livelihood activity by agencies conducting rural extension activities. Although consideration of available labour and monetary funds to invest in cattle raising is required, it was observed that income from off-farm sources may prove beneficial in providing the additional monetary funds to support cattle-raising activities and assist in providing generally poor smallholder households with enhanced economic resilience.     

Carbohydrate profiles in the graft union of young sweet cherry trees grown on dwarfing and vigorous rootstocks

In composite (scion–rootstock) dwarfing fruit trees, an overgrowth at the graft union is often observed, the severity of which is correlated with degree of dwarfing. The graft union of dwarfing sweet cherry (Prunus avium L.) rootstocks may limit soluble sugar transport or starch mobilization, leading to localized accumulation. Soluble sugars and starch were measured in the tissues surrounding the graft union of young ‘Rainier’ (2002) and ‘Lapins’ (2003) sweet cherry trees on ‘Gisela 5’ (‘Gi 5’; dwarfing) and ‘Colt’ (vigorous) rootstocks. Separate rootstock shank, rootstock, graft union, and scion tissues were analyzed for both starch and soluble sugar content throughout the growing season in both years. Starch concentrations did not vary among locations within the graft union for ‘Rainier’ on either the dwarfing or vigorous rootstock, or for ‘Lapins’ on the dwarfing stock. However, for ‘Lapins’ on ‘Colt’, starch was highest in the rootstock shank and declined vertically (rootstock shank ? rootstock > union > scion). Soluble sugar concentrations were generally similar to or higher in scions on ‘Gi 5’ than on ‘Colt’, and were similar to or lower in the rootstock and rootstock shank on ‘Gi 5’ than on ‘Colt’. Results suggest that rootstock has a significant effect of localized accumulations of carbohydrates above and within the graft union of ‘Gi 5’ and below the graft union of ‘Colt’.     

Quantitation of flavor-related alkenylbenzenes in tobacco smoke particulate by selected ion monitoring gas chromatography-mass spectrometry

Little is known about the possible health effects associated with inhaling alkenylbenzenes through cigarette smoking, even though these flavor-related compounds have known toxic effects in animals. We developed a rapid and sensitive solid-phase extraction (SPE) method to quantify seven alkenylbenzenes and piperonal in mainstream cigarette smoke particulate. The smoke particulate fraction of a single cigarette was collected on Cambridge filter pads, solvent extracted, concentrated, purified with SPE, and analyzed by selected ion monitoring gas chromatography-mass spectrometry. We positively identified and quantified five alkenylbenzenes compounds (eugenol, isoeugenol, methyleugenol myristicin, and elemicin) and piperonal in the smoke particulate from eight U.S. brands with mean levels (measured in triplicate) ranging from 6.6 to 4210 ng per cigarette. Additionally, complete blocking of nearly invisible ventilation holes in the cigarette filter increased 2- to 7-fold the percent transfer of alkenylbenzenes from tobacco to the particulate fraction of mainstream smoke.     

Modeling juvenile salmon migration using A simple markov chain

We describe movement patterns of hatchery-raised, juvenile, spring chinook salmon, Oncorhynchus tshawytscha, using a two-state Markov chain model. The existence of two states, moving and holding, is suggestedby anecdotalinformation from a large radio telemetry study; yet adequate observations of these small-scale fish behaviors are not available for estimating transition probabilities directly. Instead, we estimate the transition probability matrix from travel times within each of 11 river segments using a method of moments approach. Bootstrapped confidence intervals are presented. Results suggest that fish behavior in the region of the confluence between the Grande Ronde and Snake Rivers includes many transitions between moving and staying while fish behavior in the Snake River is more likely to include long periods of staying.     

Application of analytical standards in method development: selected chlorinated phenols and herbicides

Pure analytical standards are key components in the successful development of accurate analytical methods. When such compounds are unavailable commercially, a laboratory synthesis activity can provide such materials for use in method development. An example of such an approach is the synthesis of ethyl ethers of chlorinated phenols and ethyl esters of several phenoxy herbicides. Individual synthesis steps and the characterization of such materials are described. The use of such analytical standards in method development is vital for defining method characteristics. In addition, such materials are helpful in diagnosing chromatographic and mass spectral problems in the course of this work. Key mass spectral information including parent and daughter scans obtained from these analytical standards by using tandem mass spectrometry provide the basis for a new analytical method with increased sensitivity and specificity.