Low genetic differentiation between apotheciate Usnea florida and sorediate Usnea subfloridana (Parmeliaceae,Ascomycota) based on microsatellite data

Accurate species delimitation has a pivotal role in conservation biology, and it is especially important for threatened species where decisions have political and economic consequences. Finding and applying appropriate character sets and analytical tools to resolve interspecific relationships remains challenging in lichenized fungi. The main aim of our study was to re-assess the species boundaries between Usnea subfloridana and Usnea florida, which have been phylogenetically indistinguishable until now, but are different in reproductive mode and ecological preferences, using fungal-specific simple sequence repeats (SSR), i.e. microsatellite markers. Bayesian clustering analysis, discriminant analysis of principal components (DAPC), minimal spanning network (MSN), and principal component analysis (PCA) failed to separate U. florida and U. subfloridana populations. However, a low significant differentiation between the two taxa was observed across all populations according to AMOVA results. Also, analysis of shared haplotypes and statistical difference in clonal diversity (M) supported the present-day isolation between the apotheciate U. florida and predominantly sorediate U. subfloridana. Our results do not provide a clear support either for the separation of species in this pair or the synonymization of U. florida and U. subfloridana. We suggest that genome-wide data could help resolve the taxonomic question in this species pair.     

海洋酸化对微藻关键生理过程的调控机制及环境因素的影响

人类活动排放大量的CO₂通过海气界面进入海洋,打破原有海水碳酸盐平衡进而造成海洋酸化(OA)。OA会影响海水和海洋污染物的理化性质,进而对生活在海洋表层的浮游藻类生理过程产生显著调控作用。海洋微藻作为海洋中主要的初级生产者,其生理功能与过程的正常进行对于海洋生态系统具有重要作用。本文综述了OA对海洋微藻光合固碳、钙化过程、固氮作用3个关键生理过程的调控作用和具体机制,总结了OA条件下,环境因素(如太阳辐射、温度、营养元素)对微藻生理过程和生长的影响,以及OA通过改变典型海洋污染物(如有机污染物、重金属、微塑料)的环境行为而对微藻生理过程的调控作用。最后,结合研究现状,对未来需要开展的研究方向进行展望。本文为进一步了解OA对海洋生态系统的潜在影响提供了重要信息。     

Model-based design and control of a small-scale integrated continuous end-to-end mAb platform

A continuous integrated bioprocess available from the earliest stages of process development allows for an easier, more efficient and faster development and characterization of an integrated process as well as production of small-scale drug candidates. The process presented in this article is a proof-of-concept of a continuous end-to-end monoclonal antibody production platform at a very small scale based on a 200 ml alternating tangential flow filtration perfusion bioreactor, integrated with the purification process with a model-based design and control. The downstream process, consisting of a periodic twin-column protein A capture, a virus inactivation, a CEX column and an AEX column, was compactly implemented in a single chromatography system, with a purification time of less than 4 hr. Monoclonal antibodies were produced for 17 days in a high cell density perfusion culture of CHO cells with titers up to 1.0 mg/ml. A digital twin of the downstream process was created by modelling all the chromatography steps. These models were used for real-time decision making by the implementation of control strategies to automatize and optimize the operation of the process. A consistent glycosylation pattern of the purified product was ensured by the steady state operation of the process. Regarding the removal of impurities, at least a 4-log reduction in the HCP levels was achieved. The recovery yield was up to 60%, and a maximum productivity of 0.8 mg/ml/day of purified product was obtained.     

Reduction of medium consumption in perfusion mammalian cell cultures using a perfusion rate equivalent concentrated nutrient feed

Media preparation for perfusion cell culture processes contributes significantly to operational costs and the footprint of continuous operations for therapeutic protein manufacturing. In this study, definitions are given for the use of a perfusion equivalent nutrient feed stream which, when used in combination with basal perfusion medium, supplements the culture with targeted compounds and increases the medium depth. Definitions to compare medium and feed depth are given in this article. Using a concentrated nutrient feed, a 1.8-fold medium consumption (MC) decrease and a 1.67-fold increase in volumetric productivity (PR) were achieved compared to the initial condition. Later, this strategy was used to push cell densities above 100 × 10⁶ cells/ml while using a perfusion rate below 2 RV/day. In this example, MC was also decreased 1.8-fold compared to the initial condition, but due to the higher cell density, PR was increased 3.1-fold and to an average PR value of 1.36 g L⁻¹ day⁻¹ during a short stable phase, and versus 0.46 g L⁻¹ day⁻¹ in the initial condition. Overall, the performance improvements were aligned with the given definitions. This multiple feeding strategy can be applied to gain some flexibility during process development and also in a manufacturing set-up to enable better control on nutrient addition.     

Integration of ion exchange resin materials for a downstream-processing approach of an imine reductase-catalyzed reaction

In this study, an ion exchange resin-based downstream-processing concept for imine reductase (IRED)-catalyzed reactions was investigated. As a model reaction, 2-methylpyrroline was converted to its corresponding product (S)-2-methylpyrrolidine with >99% of conversion by the (S)-selective IRED from Paenibacillus elgii B69. Under optimized reaction conditions full conversion was achieved using a substrate concentration of 150 and 500 mmol/L of d -glucose. Seven commercially available cation- and anion-exchange resins were studied with respect to their ability to recover the product from the reaction solution. Without any pretreatment, cation-exchange resins Amberlite IR-120(H), IRN-150, Dowex Monosphere 650C, and Dowex Marathon MSC showed high recovery capacities (up to >90%). A 150-ml preparative scale reaction was performed yielding ~1 g hydrochloride salt product with >99% purity. Any further purification steps, for example, by column chromatography or recrystallization, were not required.     

Meiofauna promotes litter decomposition in stream ecosystems depending on leaf species

Litter decomposition, a fundamental process of nutrient cycling and energy flow in freshwater ecosystems, is driven by a diverse array of decomposers. As an important component of the heterotrophic food web, meiofauna can provide a trophic link between leaf‐associated microbes (i.e., bacteria and fungi)/plant detritus and macroinvertebrates, though their contribution to litter decomposition is not well understood. To investigate the role of different decomposer communities in litter decomposition, especially meiofauna, we compared the litter decomposition of three leaf species with different lignin to nitrogen ratios in litter bags with different mesh sizes (0.05, 0.25, and 2 mm) in a forested stream, in China for 78 days. The meiofauna significantly enhanced the decomposition of leaves of high‐and medium‐ quality, while decreasing (negative effect) or increasing (positive effect) the fungal biomass and diversity. Macrofauna and meiofauna together contributed to the decomposition of low‐quality leaf species. The presence of meiofauna and macrofauna triggered different aspects of the microbial community, with their effects on litter decomposition varying as a function of leaf quality. This study reveals that the meiofauna increased the trophic complexity and modulated their interactions with microbes, highlighting the important yet underestimated role of meiofauna in detritus‐based ecosystems.     

Analysis of chemometric models applied to Raman spectroscopy for monitoring key metabolites of cell culture

The Food and Drug Administration (FDA) initiative of Process Analytical Technology (PAT) encourages the monitoring of biopharmaceutical manufacturing processes by innovative solutions. Raman spectroscopy and the chemometric modeling tool partial least squares (PLS) have been applied to this aim for monitoring cell culture process variables. This study compares the chemometric modeling methods of Support Vector Machine radial (SVMr), Random Forests (RF), and Cubist to the commonly used linear PLS model for predicting cell culture components—glucose, lactate, and ammonia. This research is performed to assess whether the use of PLS as standard practice is justified for chemometric modeling of Raman spectroscopy and cell culture data. Model development data from five small-scale bioreactors (2 × 1 L and 3 × 5 L) using two Chinese hamster ovary (CHO) cell lines were used to predict against a manufacturing scale bioreactor (2,000 L). Analysis demonstrated that Cubist predictive models were better for average performance over PLS, SVMr, and RF for glucose, lactate, and ammonia. The root mean square error of prediction (RMSEP) of Cubist modeling was acceptable for the process concentration ranges of glucose (1.437 mM), lactate (2.0 mM), and ammonia (0.819 mM). Interpretation of variable importance (VI) results theorizes the potential advantages of Cubist modeling in avoiding interference of Raman spectral peaks. Predictors/Raman wavenumbers (cm⁻¹) of interest for individual variables are X1139–X1141 for glucose, X846–X849 for lactate, and X2941–X2943 for ammonia. These results demonstrate that other beneficial chemometric models are available for use in monitoring cell culture with Raman spectroscopy.     

Parameter estimation for discretely observed linear birth‐and‐death processes

Birth‐and‐death processes are widely used to model the development of biological populations. Although they are relatively simple models, their parameters can be challenging to estimate, as the likelihood can become numerically unstable when data arise from the most common sampling schemes, such as annual population censuses. A further difficulty arises when the discrete observations are not equi‐spaced, for example, when census data are unavailable for some years. We present two approaches to estimating the birth, death, and growth rates of a discretely observed linear birth‐and‐death process: via an embedded Galton‐Watson process and by maximizing a saddlepoint approximation to the likelihood. We study asymptotic properties of the estimators, compare them on numerical examples, and apply the methodology to data on monitored populations.     

粪甲虫对绢蒿荒漠春秋两季牛粪分解的影响

为明确绢蒿荒漠草地牛粪的分解规律,分别在5月(春季)和9月(秋季)设置不同堆置时间点(0、7、29、48、58 h),明确粪便理化性质变化规律,同时设置不同开口的网笼(无网笼、上下开口、上开口、全封口),探讨各生态功能类群粪甲虫对粪便分解的影响。结果表明: 春季的粪甲虫种类数显著高于秋季,秋季的粪甲虫数量显著高于春季。春季粪便的水分、全碳、全氮、全磷下降主要集中在0～29 h,堆置29 h时粪便的水分、全碳、全氮、全磷分别降低39.4%、13.9%、32.1%、26.7%。堆置58 h时粪便的中性洗涤纤维和酸性洗涤纤维分别显著降低8.0%和16.0%。秋季粪便的水分、中性洗涤纤维、酸性洗涤纤维在0～7 h内下降速度最快,堆置7 h时分别降低85.6%、10.2%和20.2%。7～58 h内中性洗涤纤维和酸性洗涤纤维上升,58 h时粪便的中性洗涤纤维和酸性洗涤纤维分别升高20.0%和13.7%。全碳、全氮和全磷的分解主要集中在0～29 h内,堆置29 h时分别降低17.5%、55.0%和64.8%。不同开口的网笼有效阻止了相应生态功能类群粪甲虫的进驻,随着粪甲虫生态功能类群的增加,粪便分解速度加快,无网笼状态下的粪便分解速度显著高于其它处理。粪甲虫种类、数量以及粪便堆置时间均显著影响牛粪的分解过程。     

《国家重点保护野生植物名录》调整的必要性、原则和程序

我国是世界上植物多样性最丰富的国家之一, 1999年发布的《国家重点保护野生植物名录》(下称《名录》(第一批))明确了国家重点保护野生植物的范围, 为依法强化保护、规范无序开发利用、提高公众保护意识奠定了基础。20多年来, 我国野生植物多样性保护形势发生了很大变化, 需要对《名录》进行调整。2018年, 国家林业和草原局、农业农村部启动《名录》调整工作, 物种的遴选遵循了5条基本原则和4条补充性原则, 这些原则主要涉及中国珍稀濒危物种, 具有重要经济、文化、科研、生态等价值物种的入选以及部分物种的排除。经国务院批准, 2021年9月7日, 国家林业和草原局、农业农村部发布了调整后的《名录》, 包括真菌类、藻类、苔藓、石松类和蕨类植物、裸子植物和被子植物, 共计约1,101种(455种和40类)野生植物列入其中。本文简要介绍了《名录》调整的必要性、原则和程序及调整后的情况。