Oncolytic reovirus synergizes with chemotherapeutic agents to promote cell death in canine mammary gland tumor

The oncolytic effects of reovirus in various cancers have been proven in many clinical trials in human medicine. Oncolytic virotherapy using reovirus for canine cancers is being developed in our laboratory. The objective of this study was to examine the synergistic anti-cancer effects of a combination of reovirus and low doses of various chemotherapeutic agents on mammary gland tumors (MGTs) in dogs. The first part of this study demonstrated the efficacy of reovirus in canine MGTs in vitro and in vivo. Reovirus alone exerted significant cell death by means of caspase-dependent apoptosis in canine MGT cell lines. A single injection of reovirus impeded growth of canine MGT tumors in xenografted mice, but was insufficient to induce complete tumor regression. The second part of this study highlighted the anti-tumor effects of reovirus in combination with low doses of paclitaxel, carboplatin, gemcitabine, or toceranib. Enhanced synergistic activity was observed in the MGT cell line treated concomitantly with reovirus and in all the chemotherapeutic agents except toceranib. In addition, combining reovirus with paclitaxel or gemcitabine at half dosage of half maximal inhibitory concentration (IC₅₀) enhanced cytotoxicity by activating caspase 3. Our data suggest that the combination of reovirus and low dose chemotherapeutic agents provides an attractive option in canine cancer therapy.     

Digital mapping of soil ecosystem services in Scotland using neural networks and relationship modelling. Part 2: Mapping of soil ecosystem services

A framework for estimating the distribution of soil ecosystem service (ES) supply is described that is based on the concept of matrix multiplication. This approach enables relationships between fundamental soil variables and associated environmental characteristics to be linked to soil processes, and hence to ecosystem functions and ecosystem services. The parameterization of these relationships was achieved using a combination of data from the Scottish Soils Database and expert knowledge. Baseline data to allow mapping of processes, functions and services across Scotland is given by digital maps of soil classes. The matrix multiplication approach constrains the relationship linkages to linear relationships and ignores potential synergies between factors at each stage, but does provide a mechanism for relating fundamental soil characteristics to ecosystem services. The approach has been tested by developing maps of selected ecosystem services in Scotland and comparing these with existing maps of the same or similar ESs. While the values and their ranges differ in each case, the spatial distribution of services is similar. The proposed mechanism is extensible at every level and can also be used to explore the impacts of land management options on environmental characteristics. This is demonstrated by using the model to estimate impacts of liming on three ecosystem services: Agricultural Capability, Carbon Sequestration and Drinking Water Provision. The model is shown to produce reasonable estimation of the impacts of this management option. Further discussion of improvements to the system and its potential applications is given.     

Detection and spread of Phytophthora austrocedri within infected Juniperus communis woodland and diversity of co‐associated Phytophthoras as revealed by metabarcoding

Phytophthora austrocedri is a recently invasive soilborne pathogen which is causing widespread mortality of Juniperus communis in northern Britain. The pathways by which a single genotype of P. austrocedri has spread to infect such a geographically dispersed range of woodland sites within a relatively short timeframe are unknown. This study examined the detectability of P. austrocedri in soil and water within infected J. communis woodland using qPCR to gain a better understanding of the pathogen's key mechanisms of spread. A Phytophthora metabarcoding method was also applied to investigate the wider diversity of Phytophthora species present in water at one of the sites. qPCR analyses of P. austrocedri in soil samples at a J. communis woodland exhibiting low‐to‐moderate levels of disease suggested a slow natural spread of the pathogen in soil, requiring high moisture conditions. However, the ubiquity of P. austrocedri DNA in soil samples collected across a heavily infected J. communis site suggests that once established at a site the pathogen can be spread readily in soil locally, most likely vectored by animal movements and/or human activities. The hypothesis that P. austrocedri is aerially transmitted in rainwater was not adequately proven, and an alternative hypothesis for the widespread distribution of the pathogen on J. communis in northern Britain is presented. Metabarcoding identified DNA from a diverse range of Phytophthora species in river and rainwater samples although the main target pathogen, P. austrocedri, was not amplified which disagreed with some of the qPCR findings. Possible reasons for this are discussed.     

Application of multiplex quantitative polymerase chain reaction methods to detect common bacterial fish pathogens in Nile tilapia,Oreochromis niloticus,hatcheries in Costa Rica

Edwardsiella spp., Streptococcus spp., and Francisella noatunensis subsp. orientalis are some of the most important fish pathogens affecting global tilapia, Oreochromis spp., aquaculture. In Costa Rica, the aquaculture industry is dominated by freshwater‐cultured Nile tilapia, Oreochromis niloticus, which are raised in all seven national provinces. At present, little is known regarding the diversity of pathogens present in these facilities, and definitive identification of agents associated with disease outbreaks are rare. To evaluate the prevalence of common bacterial pathogens in these systems, this study used multiplex quantitative polymerase chain reaction (qPCR) assays targeting Edwardsiella, Streptococcus, and Francisella species as a diagnostic and surveillance tool. In 2017, seven different tilapia hatcheries were visited, and 350 fingerlings were subjected to necropsy and molecular diagnostic evaluation. Fish exhibiting gross signs of disease were subjected to histological and microbiological analysis. For the first time, Edwardsiella anguillarum was recovered and molecularly confirmed from diseased tilapia in Costa Rica. In addition, F. noatunensis subsp. orientalis was identified in a region of Costa Rica where it had not been previously reported.     

Effects of Mosquitofish,Gambusia affinis,on Channel Catfish,Ictalurus puncatatus,Production Ponds

Mosquitofish, Gambusia sp., have been spread throughout the world to biologically control mosquitoes. However, the fish has gained a reputation as an invasive species and has been implicated in displacing native aquatic species. Gambusia affinis are native to the southeastern United States and commonly occur in commercial channel catfish, Ictalurus punctatus, production ponds. We investigated effects of mosquitofish presence on zooplankton populations, water quality, disease occurrence, and fish production in experimental ponds. There were no differences between ponds with or without mosquitofish in numbers of calanoid copepods, cyclopoid copepods, total copepods, Bosmina sp., Ceriodaphnia sp., Moina sp., Daphnia sp., or total cladocerans. There were also no differences in copepod and cladoceran sizes. Copepod nauplii were more numerous during the summer months in ponds with mosquitofish. There were no differences in water quality variables (soluble reactive phosphorus, nitrate, nitrite, ammonia, total nitrogen, total phosphorus, pH) or phytoplankton density between ponds stocked with and without mosquitofish. Catfish production and disease occurrence were also similar between ponds with and without mosquitofish. Although mosquitofish may cause problems when stocked outside their native range, there does not appear to be any adverse effects of mosquitofish presence in catfish production ponds.     

Outplanting Wyoming Big Sagebrush Following Wildfire: Stock Performance and Economics

Finding ecologically and economically effective ways to establish matrix species is often critical for restoration success. Wyoming big sagebrush (Artemisia tridentata subsp. wyomingensis) historically dominated large areas of western North America, but has been extirpated from many areas by large wildfires; its re-establishment in these areas often requires active management. We evaluated the performance (survival, health) and economic costs of container and bare-root stock based on operational plantings of more than 1.5 million seedlings across 2 200 ha, and compared our plantings with 30 other plantings in which sagebrush survival was tracked for up to 5 yr. Plantings occurred between 2001 and 2007, and included 12 combinations of stock type, planting amendment, and planting year. We monitored 10 500 plants for up to 8 yr after planting. Survival to Year 3 averaged 21% and was higher for container stock (30%) than bare-root stock (17%). Survival did not differ among container-stock plantings, whereas survival of bare-root stock was sometimes enhanced by a hydrogel dip before planting, but not by mycorrhizal amendments. Most mortality occurred during the first year after planting; this period is the greatest barrier to establishment of sagebrush stock. The proportion of healthy stock in Year 1 was positively related to subsequent survival to Year 3. Costs were minimized, and survival maximized, by planting container stock or bare-root stock with a hydrogel dip. Our results indicate that outplanting is an ecologically and economically effective way of establishing Wyoming big sagebrush. However, statistical analyses were limited by the fact that data about initial variables (stock quality, site conditions, weather) were often unrecorded and by the lack of a replicated experimental design. Sharing consistent data and using an experimental approach would help land managers and restoration practitioners maximize the success of outplanting efforts.     

Abiotic factors limiting photosynthesis in Abies lasiocarpa and Picea engelmannii seedlings below and above the alpine timberline

Most research on the occurrence and stability of alpine timberlines has focused on correlations between adult tree growth and mean temperatures rather than on specific mechanisms. Timberline migration to higher altitude is dependent on new seedling establishment in the tree-line ecotone; however, reductions in photosynthetic carbon gain in establishing seedlings have previously been interpreted solely in terms of decreased seedling survival. Our objective was to evaluate the impact of abiotic factors (temperature, light and water) on photosynthetic carbon gain in young seedlings of the two dominant conifer tree species occurring naturally above (tree-line ecotone site, TS) and below (forest site, FS) a Rocky Mountain timberline in southeastern Wyoming, USA. Coincidentally, measurements were made during an unusually dry summer. Mean daily photosynthesis in seedlings of both Abies lasiocarpa (Hook.) Nutt. (subalpine fir) and Picea engelmannii Parry ex Engelm. (Engelmann spruce) was less at TS than at FS (19 and 29%, respectively). Minimum nighttime temperatures below 2 degrees C were more frequent at TS than at FS and were associated with reduced maximum photosynthesis the following day. Low midday water potentials were associated with a reduction in carbon gain at both sites early in the season, prior to snowmelt, as well as late in the season when soils began to dry. However, the lower photosynthetic rates at TS than at FS appeared to be unrelated to seedling water status because seedlings at both sites had similar xylem pressure potentials. Solar irradiance was highly variable at both sites as a result of uneven shading by neighboring trees, although this variation was substantially reduced on cloudy days (44% of all days observed). Compared with sunny days, cloudy days resulted in greater integrated daily carbon gain at both sites (41% increase at TS and 69% increase at FS), based on a simulated photosynthesis model. Photosynthetic responses to temperature, sunlight and water suggest that variable solar irradiance and nighttime temperatures were major abiotic factors limiting photosynthetic carbon acquisition in these young seedlings, especially for seedlings growing in the tree-line ecotone.     

Cost-benefit tradeoffs in engineered lac operons

Cells must balance the cost and benefit of protein expression to optimize organismal fitness. The lac operon of the bacterium Escherichia coli has been a model for quantifying the physiological impact of costly protein production and for elucidating the resulting regulatory mechanisms. We report quantitative fitness measurements in 27 redesigned operons that suggested that protein production is not the primary origin of fitness costs. Instead, we discovered that the lac permease activity, which relates linearly to cost, is the major physiological burden to the cell. These findings explain control points in the lac operon that minimize the cost of lac permease activity, not protein expression. Characterizing similar relationships in other systems will be important to map the impact of cost/benefit tradeoffs on cell physiology and regulation.     

Beta-lactam antibiotics prevent Salmonella-mediated bovine encephalopathy regardless of the β-lactam resistance status of the bacteria

This study assessed the capacity of β-lactam antibiotics to prevent salmonella-mediated encephalopathy in calves given the putative neuroprotective effects of these drugs of increasing glutamate export from the brain. Both ampicillin and ceftiofur prevented the development of encephalopathy despite resistance of the inoculated Salmonella enterica serovar Saint-Paul isolate to both drugs. A glutamate receptor antagonist also prevented this salmonella-mediated encephalopathy. Glutamate exporters were hyper-expressed in the presence of β-lactam antibiotics while a glutamate export inhibitor obviated the effects of these antibiotics, demonstrating a neuroprotective effect through glutamate export from the brain. The findings indicate that β-lactam antibiotics remain an important treatment option for this atypical form of bovine salmonellosis.