Amino-modified polystyrene nanoplastics induced multiple response of Artemia hemocytes.

Polystyrene polymers cause severe toxicity to aquatic animals. However, the process and mechanisms of innate immunity of invertebrates living at the bottom of the food chain to these pollutants remain unclear. In this study, the blood system responses of zooplankton Artemia were assessed through in vivo and in vitro exposure to amino-modified polystyrene nanoplastics (PS-NH2 NPs). The results indicated that the LC50 values of PS-NH2 NPs were 1.09 µg·mL-1 over 48 h and 0.42 µg·mL-1 over 7 d. Based on the five hemocyte subpopulations identified in Artemia, in vitro exposure assays revealed that phagocytosis was performed by plasmocytes and granulocytes with phagocytic rate of 22.64 %. TEM analysis further showed that PS-NH2 NPs caused cytoplasm vacuolization, swollen mitochondria, and lipid processing disorder. Gene expression pattern results demonstrated that Spatzle, Tollip, Hsp70, Hsp90, Casp8, API5and Pxn were significantly upregulated upon acute and chronic exposure (p < 0.05), while chronic exposure could induce significantly upregulation of ProPO (p < 0.05). Moreover, PS-NH2 NPs exposure remarkably varied the hemolymph microbiota and hemogram, particularly by increasing the proportion of adipohemocytes and phagocytes (p < 0.05). Our findings suggest that PS-NH2 NPs induce different responses in Artemia hemocyte, as primarily reflected by phagocytic processes, expression of immune and apoptosis relating genes, cell fates, hemogram and hemolymph microbiota variations. These findings support the possibility of using Artemia hemocytes as bioindicator to estimate nanoplastics pollution, thus contributing to hematological toxicity research in response to nanoplastics.

Spatio-temporal patterns of ovarian development and VgR gene silencing reduced fecundity in parthenogenetic Artemia.

The halophilic zooplankton brine shrimp Artemia has been used as an experimental animal in multidisciplinary studies. However, the reproductive patterns and its regulatory mechanisms in Artemia remain unclear. In this study, the ovarian development process of parthenogenetic Artemia (A. parthenogenetica) was divided into five stages, and oogenesis or egg formation was identified in six phases. The oogenesis mode was assumed to be polytrophic. We also traced the dynamic translocation of candidate germline stem cells (cGSCs) using EdU labelling and elucidated several key cytological events in oogenesis through haematoxylin and eosin staining and fluorescence imaging. Distinguished from the ovary structure of insects and crustaceans, Artemia germarium originated from ovariole buds and are located at the base of the ovarioles. RNA-seq based on five stages of ovarian development identified 2657 upregulated genes related to reproduction by pair-to-pair comparison. Gbb, Dpp, piwi, vasa, nanos, VgA and VgR genes associated with cGSCs recognition and reproductive development were screened and verified using qPCR. Silencing of the VgR gene in A. parthenogenetica (Ap-VgR) at ovarian development Stage II led to a low level of gene expression (less than 10%) within 5 days, which resulted in variations in oogenesis-related gene expression and significantly inhibited vitellogenesis, impeded oocyte maturation, and eventually decreased the number of offspring. In conclusion, we have illustrated the patterns of ovarian development, outlined the key spatio-temporal features of oogenesis and identified the negative impacts of VgR gene knockdown on oogenesis using A. parthenogenetica as an experimental animal. The findings of this study also lay a foundation for the further study of reproductive biology of invertebrates.

Sustainable urban system structure evaluation in sparsely populated areas: case study of the Qinghai-Tibet Plateau in China.

Urbanization in sparsely populated areas is critical for sustainability. The Qinghai-Tibet Plateau is a typical example of an ecologically fragile region that plays a crucial role in China's ecological safety and water resource protection. We use a social network analysis to illustrate the structure of the urban system on the plateau and find that the agglomeration and diffusion capabilities of the core nodes are weak, which presents an obstacle to the sustainable development of the urban system. Nevertheless, we find that the intermediate nodes-which serve multiple integration functions for the various cultures, ethnic groups, and religions in the region-are core nodes that divide the dispersed cities and towns into four subgroups that function as small worlds (The cities and towns within one subgroup connect closely and each subgroup is organized independently somehow from others). Based on this finding, we suggest implementing a "double-layer" urban system to promote the sustainable urban development of the Qinghai-Tibet Plateau. This "double-layer" system breaks the usual urbanization trend, in which cities grow in size and their distribution becomes more concentrated; instead it provides a feasible way to maintain urban sustainability in a sparsely populated area.