Engineering challenges in high density cell culture systems

High density cell culture systems offer the advantage of production of bio-pharmaceuticals in compact bioreactors with high volumetric production rates; however, these systems are difficult to design and operate. First of all, the cells have to be retained in the bioreactor by physical means during perfusion. The design of the cell retention is the key to performance of high density cell culture systems. Oxygenation and media design are also important for maximizing the cell number. In high density perfusion reactors, variable cell density, and hence the metabolic demand, require constant adjustment of perfusion rates. The use of cell specific perfusion rate (CSPR) control provides a constant environment to the cells resulting in consistent production. On-line measurement of cell density and metabolic activities can be used for the estimation of cell densities and the control of CSPR. Issues related to mass transfer and mixing become more important at high cell densities. Due to the difference in mass transfer coefficients for oxygen and CO₂, a significant accumulation of dissolved CO₂ is experienced with silicone tubing aeration. Also, mixing is observed to decrease at high densities. Base addition, if not properly done, could result in localized cell lysis and poor culture performance. Non-uniform mixing in reactors promotes the heterogeneity of the culture. Cell aggregation results in segregation of the cells within different mixing zones. This paper discusses these issues and makes recommendations for further development of high density cell culture bioreactors.     

Particle size influences decay rates of environmental DNA in aquatic systems

Environmental DNA (eDNA) analysis is a powerful tool for remote detection of target organisms. However, obtaining quantitative and longitudinal information from eDNA data is challenging, requiring a deep understanding of eDNA ecology. Notably, if the various size components of eDNA decay at different rates, and we can separate them within a sample, their changing proportions could be used to obtain longitudinal dynamics information on targets. To test this possibility, we conducted an aquatic mesocosm experiment in which we separated fish-derived eDNA components using sequential filtration to evaluate the decay rate and changing proportion of various eDNA particle sizes over time. We then fit four alternative mathematical decay models to the data, building towards a predictive framework to interpret eDNA data from various particle sizes. We found that medium-sized particles (1–10 μm) decayed more slowly than other size classes (i.e., <1 and > 10 μm), and thus made up an increasing proportion of eDNA particles over time. We also observed distinct eDNA particle size distribution (PSD) between our Common carp and Rainbow trout samples, suggesting that target-specific assays are required to determine starting eDNA PSDs. Additionally, we found evidence that different sizes of eDNA particles do not decay independently, with particle size conversion replenishing smaller particles over time. Nonetheless, a parsimonious mathematical model where particle sizes decay independently best explained the data. Given these results, we suggest a framework to discern target distance and abundance with eDNA data by applying sequential filtration, which theoretically has both metabarcoding and single-target applications.     

Integrating the spatial proximity effect into the assessment of changes in ecosystem services for biodiversity conservation

The assessment of the value of ecosystem services is a valuable tool for biodiversity conservation that can facilitate better environmental policy decision-making and land management, and can help land managers develop interventions to compensate for biodiversity loss at the patch level. Previous studies have suggested that it is appropriate to assess the value of biodiversity for conservation planning by considering both the condition of the landscape and the spatial configuration of adjacent land uses that can be reflected as a proximity effect. This research examines the influence of spatial proximity on biodiversity conservation from the ecosystem service perspective based on the assumption that the variation in the proximity effect caused by land cover change has positive or negative impacts on ecological services. Three factors related to the spatial characteristics of the landscape were considered in this approach: the relative artificiality of the land cover types, the distance decay effect of patches and the impact of one land cover type on others. The proximity effect change (PE_C) parameter reflected the relationship between the spatial proximity effect and biodiversity conservation. The results of a quantitative and spatial comparative analysis of the proposed method and the conventional method in Yingkou for the periods of 2000–2005 and 2005–2010 showed that the former can account for the temporal and spatial changes in ecosystem services for biodiversity conservation that were caused by patch-level changes as well as the interaction between the altered and adjacent patches from a spatial perspective. The metric can also identify the most critical areas for biodiversity protection and inform the efficient allocation of limited land resources for nature conservation to maximize the benefit to biodiversity by guiding the process of land-use change, particularly urbanization and agriculture. Future studies should focus on the other important factors that are applicable to the assessment of the value of biodiversity conservation in socio-ecological systems, where society and nature are mutually capable of fulfilling their roles.     

国家种质库部分作物种子生活力监测结果与分析

对国家种质库中23种作物1.8万多份种子进行了生活力监测,结果表明,贮存10～12年后,96%以上的被监测种子发芽率仍保持在85%以上,但有95份种子(占被监测份数的0.51%)出现了明显的下降(发芽率从80%以上降至70%以下).胡萝卜、莴苣、棉花、胡麻、蓖麻等5种作物种子整体监测发芽率平均值都出现了显著下降,而胡萝卜、莴苣这二种作物的监测发芽率下降更为显著.不同起始发芽率水平及贮存前所处环境条件对贮存种子的生活力下降有一定影响.     

Impending disaster or sliver of hope for Southeast Asian forests? The devil may lie in the details

Here, I report on how forest area in Southeast Asia has changed for different types of forest and across different countries between the periods of 1990–2000 and 2000–2005. The loss of old growth forests has accelerated in Indonesia, Cambodia and Vietnam but have ceased in Thailand, Malaysia, Laos and the Philippines. Secondary forests continue to be lost in Malaysia, Cambodia, Laos, Myanmar, Indonesia, Thailand and the Philippines. Plantation forests have increased in area by 25.0% from 1990 to 2005 but still comprise only 6.2% of the total forest area in Southeast Asia. Overall, the loss of native forests (old growth and secondary forests) has slowed down in Thailand, the Philippines and Brunei but has worsened in Laos, Myanmar, Indonesia, Malaysia and Cambodia.     

Effect of Environmental Factors on Cyanobacteria Richness in Some Agricultural Soils

The physicochemical variations of soil, such as temperature, pH, nutrients, and the type of plant cultivation, affect the diversity of cyanobacteria, whether heterocystous or not. The aim of this study was to identify the species of cyanobacteria in a soil and the effect of environmental characteristics on cyanobacteria. Soil samples collected from six different agricultural sites in Al Diwaniyah city/Iraq during September 2016 in the autumn season were analyzed, and the physicochemical characteristics of the samples were measured using approved methods.

The results showed significant correlation and differences between cyanobacteria composition, distribution, and physicochemical factors among soil sites. The Agricultural soil was slightly alkaline and moderately saline and contained abundant nutrients, cations and a high percentage of organic matter. All these characteristics influenced the distribution and diversity of cyanobacteria. Ninety-six species were identified, including four heterocystous species represented by Anabaena, Calothrix, Cylnidrospermum, and Nostoc. However, the non-heterocystous were represented by 13 species: Aphanocapsa, Aphanothece, Arthrospira, Chroococcus, Gloeocapsa, Lyngbya, Merismopedia, Microcystis, Microcoleus, Oscillatoria, Phormidium, Schizothrix, and Spirulina. The dominant species of cyanobacteria was Oscillatoria, followed by Phormidium, Chroococcus, Gleocapsa and Lyngbya. The highest value of Shannon’s and Simpson’s diversities were registered in the Ghammas site, which is a paddy field, but the lowest was registered in the Afak site, cultivated with the alfalfa plant. Soil was classified as finely textured with silty clayey characterization, favorable for cyanobacteria growth.     

Helianthus maximiliani and species fine‐scale spatial pattern affect diversity interactions in reconstructed tallgrass prairies

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Estimating site occupancy and detectability of the threatened partridge pigeon (Geophaps smithii) using camera traps

Since European settlement, many granivorous birds of northern Australia's savanna landscapes have declined. One such example, the partridge pigeon (Geophaps smithii), has suffered a significant range contraction, disappearing from at least half of its pre‐European range. Multiple factors have been implicated in this decline, including the loss of traditional Aboriginal burning practices, grazing by large exotic herbivores and predation by feral cats (Felis catus). While populations of partridge pigeon on the Tiwi Islands may be particularly important for the long‐term persistence of this species, they too may be at risk of decline. However, as a reliable method to detect this species has not yet been developed and tested, we lack the ability to identify, at an early stage, the species' decline in a given location or region. This severely limits our capacity to make informed management decisions. Here, we demonstrate that the standard camera trapping approach for native mammal monitoring in northern Australia attained an overall probability of detecting partridge pigeon greater than 0.98. We thus provide a robust estimate of partridge pigeon site occupancy (0.30) on Melville Island, the larger of the two main Tiwi Islands. The information presented here for the partridge pigeon represents a critical first step towards the development of optimal monitoring programmes with which to gauge population trajectories, as well as the response to remedial management actions. In the face of ongoing biodiversity loss, such baseline information is vital for management agencies to make informed decisions and should therefore be sought for as many species as possible.     

Tricin‐lignins: occurrence and quantitation of tricin in relation to phylogeny

Tricin [5,7‐dihydroxy‐2‐(4‐hydroxy‐3,5‐dimethoxyphenyl)‐4H‐chromen‐4‐one], a flavone, was recently established as an authentic monomer in grass lignification that likely functions as a nucleation site. It is linked onto lignin as an aryl alkyl ether by radical coupling with monolignols or their acylated analogs. However, the level of tricin that incorporates into lignin remains unclear. Herein, three lignin characterization methods: acidolysis; thioacidolysis; and derivatization followed by reductive cleavage; were applied to quantitatively assess the amount of lignin‐integrated tricin. Their efficiencies at cleaving the tricin‐(4′–O–β)‐ether bonds and the degradation of tricin under the corresponding reaction conditions were evaluated. A hexadeuterated tricin analog was synthesized as an internal standard for accurate quantitation purposes. Thioacidolysis proved to be the most efficient method, liberating more than 91% of the tricin with little degradation. A survey of different seed‐plant species for the occurrence and content of tricin showed that it is widely distributed in the lignin from species in the family Poaceae (order Poales). Tricin occurs at low levels in some commelinid monocotyledon families outside the Poaceae, such as the Arecaceae (the palms, order Arecales) and Bromeliaceae (Poales), and the non‐commelinid monocotyledon family Orchidaceae (Orchidales). One eudicotyledon was found to have tricin (Medicago sativa, Fabaceae). The content of lignin‐integrated tricin is much higher than the extractable tricin level in all cases. Lignins, including waste lignin streams from biomass processing, could therefore provide a large and alternative source of this valuable flavone, reducing the costs, and encouraging studies into its application beyond its current roles.     

Behavioral Phenotyping of Murine Disease Models with the Integrated Behavioral Station (INBEST)

Due to rapid advances in genetic engineering, small rodents have become the preferred subjects in many disciplines of biomedical research. In studies of chronic CNS disorders, there is an increasing demand for murine models with high validity at the behavioral level. However, multiple pathogenic mechanisms and complex functional deficits often impose challenges to reliably measure and interpret behavior of chronically sick mice. Therefore, the assessment of peripheral pathology and a behavioral profile at several time points using a battery of tests are required. Video-tracking, behavioral spectroscopy, and remote acquisition of physiological measures are emerging technologies that allow for comprehensive, accurate, and unbiased behavioral analysis in a home-base-like setting. This report describes a refined phenotyping protocol, which includes a custom-made monitoring apparatus (Integrated Behavioral Station, INBEST) that focuses on prolonged measurements of basic functional outputs, such as spontaneous activity, food/water intake and motivated behavior in a relatively stress-free environment. Technical and conceptual improvements in INBEST design may further promote reproducibility and standardization of behavioral studies.