Effects of three-dimensional spheroid culture on equine mesenchymal stem cell plasticity

Mesenchymal stem cells (MSCs) are useful candidates for tissue engineering and cell therapy fields. We optimize culture conditions of equine adipose tissue-derived MSCs (eAD-MSCs) for treatment of horse fractures. To investigate enhancing properties of three-dimensional (3D) culture system in eAD-MSCs, we performed various sized spheroid formation and determined changes in gene expression levels to obtain different sized spheroid for cell therapy. eAD-MSCs were successfully isolated from horse tailhead. Using hanging drop method, spheroid formation was generated for three days. Quantitative real-time PCR was performed to analyze gene expression. As results, expression levels of pluripotent markers were increased depending on spheroid size and the production of PGE₂ was increased in spheroid formation compared to that in monolayer. Ki-67 showed a remarkable increase in the spheroid formed with 2.0?×?10⁵ cells/drop as compared to that in the monolayer. Expression levels of angiogenesis-inducing factors such as VEGF, IL-6, IL-8, and IL-18 were significantly increased in spheroid formation compared to those in the monolayer. Expression levels of bone morphogenesis-inducing factors such as Cox-2 and TGF-β1 were also significantly increased in spheroid formation compared to those in the monolayer. Expression levels of osteocyte-specific markers such as RUNX2, osteocalcin, and differentiation potential were also significantly increased in spheroid formation compared to those in the monolayer. Therefore, spheroid formation of eAD-MSCs through the hanging drop method can increases the expression of angiogenesis-inducing and bone morphogenesis-inducing factors under optimal culture conditions.     

Weeds,aphids, and specialist parasitoids and predators benefit differently from organic and conventional cropping of winter cereals

The aphid–natural enemy interaction in winter wheat fields constitutes a complex system that has been frequently studied because of its implication for biological control. However, not all of the aphids living in cereal fields are crop pests, as there are also aphids living on weeds that may serve as alternative hosts or prey for aphid parasitoids or predators. In this context, a concomitant survey of the plant and insect communities was conducted to understand how different plant communities affect the abundance and richness of aphids and the interactions with their natural enemies. The plant community was split into functional groups (grasses, legumes and forbs), and the aphid community was divided into feeding groups according to their host preferences (specialists in grasses or forbs). The grass aphids, which dominated the total aphid catches, responded positively to grass cover, which was particularly enhanced in the conventional fields. Conversely, the forb aphids, which mainly conditioned the total species richness of the aphids, were closely correlated with the local abundance of legumes. The system of cereal aphid-parasitoids was enhanced in the conventional fields, where the abundance of grasses was higher, whereas the legumes of the organic fields indirectly played a key role in enhancing the richness of the parasitoids and the abundance of predators. Our findings indicate that a bottom-up effect exists throughout the plant community, aphids, and aphidophagous insects and that plant community characteristics should be considered to better understand cereal aphid control.     

Analysis of a site x year experiment with timothy polycross progeny

Summary Results from a cooperative breeding programme with timothy for the northern areas of Scandinavia are presented. The main aim of the programme was to identify genotypes for synthetic populations that are high yielding, adapted over the whole area and stable over years. A polycross comprising 12 genotypes from each of five sites within the region was formed. The subsequent 60 half-sib families and four reference varieties were then compared under sward conditions at the same sites. The trials lasted for three years and results for total dry matter yields are presented.Significant differences in yield between lines were found. There were also significant two and three factor interactions between lines, sites and years. A new two-step procedure is presented to select the genotypes to make up a synthetic variety. Firstly, a superiority measure (Q-value) is used to select a group of genotypes that are high yielding and stable over sites and years. Secondly, the GxE part of the Q-value is split into two terms. One measures adaptation to predictable environmental conditions. The other one measures stability to unpredictable changes in the environment. A plot of these two measures provides a tool for discarding unstable or poorly adapted genotypes. The statistical properties of the selection parameters are discussed.     

Estimating the spatial scales of landscape effects on abundance

Context

Spatial variation in abundance is influenced by local- and landscape-level environmental variables, but modeling landscape effects is challenging because the spatial scales of the relationships are unknown. Current approaches involve buffering survey locations with polygons of various sizes and using model selection to identify the best scale. The buffering approach does not acknowledge that the influence of surrounding landscape features should diminish with distance, and it does not yield an estimate of the unknown scale parameters.

Objectives

The purpose of this paper is to present an approach that allows for statistical inference about the scales at which landscape variables affect abundance.

Methods

Our method uses smoothing kernels to average landscape variables around focal sites and uses maximum likelihood to estimate the scale parameters of the kernels and the effects of the smoothed variables on abundance. We assessed model performance using a simulation study and an avian point count dataset.

Results

The simulation study demonstrated that estimators are unbiased and produce correct confidence interval coverage except in the rare case in which there is little spatial autocorrelation in the landscape variable. Canada warbler abundance was more highly correlated with site-level measures of NDVI than landscape-level NDVI, but the reverse was true for elevation. Canada warbler abundance was highest when elevation in the surrounding landscape, defined by an estimated Gaussian kernel, was between 1300 and 1400 m.

Conclusions

Our method provides a rigorous way of formally estimating the scales at which landscape variables affect abundance, and it can be embedded within most classes of statistical models.

     

Feeding behavior under dark conditions in larvae of sutchi catfish <Emphasis Type="Italic">Pangasianodon hypophthalmus</Emphasis>

Sutchi catfish Pangasianodon hypophthalmus hatch with morphologically immature sensory organs; however, sensory organs develop rapidly with larval growth. Two-day-old larvae commenced ingesting Artemia nauplii. The larvae displayed many taste buds on the barbels, the head surface, and in the buccal cavity. Other sense organs were also well developed at this stage. Feeding experiments revealed that 2-day-old larvae ingested Artemia under both light and dark conditions, moreover, the larvae could ingest frozen dead Artemia. The ingestion rates for 4- and 7-day-old larvae were significantly higher under dark conditions than under light conditions. The rates using frozen dead Artemia were mostly higher than the rates using live Artemia. Therefore, feeding behavior under dark conditions is most likely not mediated by visual or mechanical senses, but rather by chemosensory senses, such as taste buds. Larval fish are vulnerable to predators; thus, if they can search for and eat food at night, they can avoid diurnal predators. The behavior observed here appears to represent their survival strategy. Moreover, these results suggest a new possibility that sutchi catfish larvae can be reared under dark or dim light conditions in order to improve survival and growth rates as in the case of African catfish Clarias gariepinus.     

A NMR study of water distribution in hardwoods at several equilibrium moisture contents

The water state of one tropical (Robinia coccinea) and two temperate (Acer saccharum and Fagus grandifolia) hardwoods was determined at different equilibrium moisture contents (EMC) during desorption at 25°C. NMR technique was used to separate different components of water in wood. The species studied presented different structures, which were apparent on the spin–spin relaxation T₂ values. Three different water components were separated: slow T₂ (liquid water in vessel elements), medium T₂ (liquid water in fiber and parenchyma elements) and fast T₂ (bound or cell wall water). The NMR results showed that even at equilibrated conditions a region exists where loss of liquid water and bound water takes place simultaneously. This region will vary according to the wood structure. Finally, liquid water was present at EMC lower than the fiber saturation point, which contradicts the concept of this point when considered as a bulk property of wood.     

Effects of a plastic cover on soil moisture change in a Mediterranean climatic regime

Water deficit is a serious problem for most agricultural crops, especially in arid–semiarid regions, and limits sustainable development. Production can be improved by reducing evapotranspiration and loss of infiltrated water by the use of a plastic cover. We monitored soil moisture fortnightly over 1 yr using a neutron probe near four trees in an olive grove (Olea europaea, var. Arbequina), two of which had a plastic cover buried in the topsoil around them. These trees were monitored using three neutron probe access tubes per tree to compare the behaviour of soil moisture over time with two other identically instrumented and nearby trees with no cover. Analysis was based on the resultant moisture profiles. The plastic cover retained moisture and increased soil water residence time. During the dry season, the amount of water retained in the plots was at most 15–20% greater in the mid‐season and at least 5–6% greater at the end of season than in the central part of the plots near the trunk. The plastic cover was effective to ca. 50 cm with maximum water content near the soil surface. During the wet season, the cover did not affect soil water. Soil moisture was greater near the tree trunk as a result of stemflow and throughfall.     

Relating tradable credits for biodiversity to sustainability criteria in a dynamic landscape

Tradable biodiversity credit systems provide flexible means to resolve conflicts between development and conservation land-use options for habitats occupied by threatened or endangered species. We describe an approach to incorporate the influence of habitat fragmentation into the conservation value of tradable credits. Habitat fragmentation decreases gene flow, increases rates of genetic drift and inbreeding, and increases probabilities of patch extinction. Importantly, tradable credit systems will change the level of fragmentation over time for small and/or declining populations. We apply landscape equivalency analysis (LEA), a generalizable, landscape-scale accounting system that assigns conservation value to habitat patches based on patch contributions to abundance and genetic variance at landscape scales. By evaluating habitat trades using two models that vary the relationship between dispersal behaviors and landscape patterns, we show that LEA provides a novel method for limiting access to habitat at the landscape-scale, recognizing that the appropriate amount of migration needed to supplement patch recruitment and to offset drift and inbreeding will vary as landscape pattern changes over time. We also found that decisions based on probabilities of persistence alone would ignore changes in migration, genetic drift, and patch extinction that result from habitat trades. The general principle of LEA is that habitat patches traded should make at least equivalent contributions to rates of recruitment and migration estimated at a landscape scale. Traditional approaches for assessing the “take” and “jeopardy” standards under the Endangered Species Act based on changes in abundance and probability of persistence may be inadequate to prevent trades that increase fragmentation.     

Response of Thawed Epidi dymal Red Deer Spermatozoa to Increasing Concentrations of Hydrogen Peroxide,and Importance of Individual Male Variability

Oxidative stress represents a challenge during sperm manipulation. We have tested the effect of increasing hydrogen peroxide (H₂O₂) levels on red deer spermatozoa after cryopreservation, and the role of male‐to‐male variation in that response. In a first experiment, eight thawed samples were submitted to 0, 25, 50, 100 and 200 μm H₂O₂ for 2 h at 37°C. Intracellular reactive oxygen species (H₂DCFDA‐CM) increased with H₂O₂ concentration, but we only detected a decrease in sperm function (motility by CASA and chromatin damage by sperm chromatin structure assay) with 200 μm . Lipoperoxidation assessed by the thiobarbituric acid reactive substance (TBARS) method increased slightly with 50 μm H₂O₂ and above. In a second experiment, samples from seven males were submitted to 0 and 200 μm H₂O₂ for 2 h, triplicating the experiment within each male. Males differed at thawing and regarding their response to incubation and H₂O₂ presence. We found that the kinematic parameters reflected male‐to‐male variability, whereas the response of the different males was similar for lipid peroxidation and viability. A multiparametric analysis showed that males grouped differently if samples were assessed after thawing, after incubation without H₂O₂ or after incubation with H₂O₂. Red deer spermatozoa are relatively resilient to H₂O₂ after thawing, but it seems to be a great male‐to‐male variability regarding the response to oxidative stress. The acknowledgement of this individual variability might improve the development of optimized sperm work protocols.     

Bioprospection of Microalgae and Cyanobacteria as Biocontrol Agents Against Vibrio campbellii and Their Use in White Shrimp Litopenaeus vannamei Culture

Vibrio spp. are the most common and harmful shrimp pathogenic bacteria; however, microalgae and cyanobacteria have the ability to produce antimicrobial substances against these species. In this study, the organic and aqueous extracts of 28 species of marine microalgae and cyanobacteria were screened against Vibrio campbellii M1. Two of these phytoplankton species with antibacterial activity in aqueous extracts (Dunaliella tertiolecta and Skeletonema costatum) and nontoxic to brine shrimp Artemia franciscana nauplii were used to evaluate their anti‐Vibrio effect when used as green‐water cultures in Vibrio‐challenged white shrimp Litopenaeus vannamei cultures. No differences in mortality of juvenile L. vannamei were observed between treatments tested, suggesting that the pathogenicity of V. campbellii could be related to the growth stage of shrimp. The proximal composition of D. tertiolecta and S. costatum was in the recommended range for penaeid shrimp nutrition, allowing shrimp supplemented with these microalgae to have significantly greater total length and weight than control shrimp. Shrimp supplemented with S. costatum presented the highest values of organic mass (11.48 mg/organism) and growth rate (0.31 mg/d) in comparison to D. tertiolecta. These results indicate that microalgae are not only capable of producing antibacterial compounds against Vibrio but can also help shrimp nutrition.