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Spatio-temporal patterns and processes of genetic differentiation in passively dispersing zooplankton are drawing much attention from both ecologists and evolutionary biologists. Two opposite phylogeographical scenarios have already been demonstrated in rotifers, which consist of high levels of genetic differentiation among populations even on small geographical scales on the one hand and the traditionally known cosmopolitanism that is associated with high levels of gene flow and long-distance dispersal via diapausing stages on the other hand. Here, we analysed the population genetic structure and the phylogeography of the Brachionus calyciflorus species complex in eastern China. By screening a total of 318 individuals from ten locations along a 2320-km gradient and analysing samples from two growing seasons, we aimed at focusing on both small- and large-scale patterns. We identified eight cryptic species and verified species status of two of these by sexual reproduction tests. Samples in summer and winter yielded different cryptic species. The distribution patterns of these genetically distinct cryptic species were diverse across eastern China, from full cosmopolitanism to local endemism. The two most abundant cryptic species BcWIII and BcSW showed a pattern of strong genetic differentiation among populations and no significant isolation by distance. Long-distance colonization, secondary contact and recent range expansion are probably responsible for the indistinct pattern of isolation by distance. Our results suggest that geographical distance is more important than temporal segregation across seasons in explaining population differentiation and the occurrence of cryptic species. We explain the current phylogeographical structure in the B. calyciflorus species complex by a combination of recent population expansion, restricted gene flow, priority effects and long-distance colonization.

Worldwide, there have been few comparative studies on rotifer communities in subtropical lakes. We studied changes in rotifer community structure over 1 year and its relationship to several physicochemical variables in five subtropical shallow lakes in East China, covering a nutrient gradient from mesotrophy to moderate eutrophy. In these lakes, the genera Brachionus, Lecane, and Trichocerca dominated the rotifer species composition, and Polyarthra dolichoptera, Keratella cochlearis, Filinia longiseta, T. pusilla, and Anuraeopsis fissa were the dominant species. With increased nutrient loading, total rotifer abundance and species dominance increased, indicating that rotifer abundance might be a more sensitive indicator of trophic state than species composition. Comparative analyses of the six rotifer community indices calculated in this study and redundancy analysis (RDA) revealed that the two slightly eutrophic lakes and the other two moderately eutrophic lakes exhibited a high degree similarity in community structure. This suggests that the trophic state of a lake determines the rotifer community structure. In contrast, in the two moderately eutrophic lakes, the mass ratios of TN:TP and the contents of TP suggested N-limitation and cyanobacteria dominance in phytoplankton communities might be possible. In these lakes TN played a more important role in shaping the rotifer community according to stepwise multiple regression and RDA. RDA analysis also suggested that rotifer species distribution was strongly associated with trophic state and water temperature, with water temperature being the most important factor in determining seasonality.

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Heat stress will stimulate cells of living organisms to generate heat shock proteins (Hsps). In the mouse liver, impacts of heat stress on hepatocyte proliferation, apoptosis and metabolism have not been studied systematically at different temperatures. In this research, the test mice were heated to 40, 42, 44 and 46°C, respectively, for 20 min and recovered at room temperature for 8 h in normal feeding conditions; the control animals were kept at room temperature without heat stress. The expression levels of Hsp70, Pcna, Bax, Bcl2, cytochrome P450 1A2 (CYP1A2), CYP2E1 and analog of CYP3A4 (not reported in mouse before), the parameters reflecting stress strength, cell proliferation, apoptosis and metabolism, were detected by western blotting, immunohistochemistry and semi-quantitative RT-PCR in test and control mice. Haematoxylin-eosin (H&E) staining and TUNEL analysis were further used to study the impacts of heat stress at different temperatures on hepatocellular necrosis and apoptosis. Serum AST and ALT levels, the markers of liver injury, were measured after heat stress at different temperatures. The data show that Hsp70 expression was significantly increased when temperature increased (P < 0.05). At lower temperatures (40 or 42°C), expression of Pcna, CYP1A2 and analog of CYP3A4 were considerably increased (P < 0.05) while hepatocyte necrosis and apoptosis were not induced (P > 0.05). At higher temperatures (44 or 46°C), expression of Pcna was decreased while hepatocyte necrosis and apoptosis were induced (P < 0.05). Expressions of CYP1A2 and analog of CYP3A4 were decreased especially at 46°C (P < 0.05). Expression of CYP2E1 could not be detected to increase at 40°C but was at high levels at 42, 44 and 46°C (P < 0.05). Expressions of AST and ALT were not different between the test mice and control mice at 40°C while they were significantly higher in the test mice than those in the control mice at 42 (P < 0.05), 44 and 46°C (P < 0.01). In conclusion, heat stress at lower temperatures promotes hepatocyte proliferation and improves the metabolic efficiency in mouse liver while heat stress at higher temperatures inhibits hepatocyte proliferation, promotes hepatocyte apoptosis and induces hepatocyte necrosis. This may give a hint to understanding human liver injury in high temperatures. Moreover, it is the first time that the analog of CYP3A4 was detected in mouse hepatocellular cytoplasm. It is worthwhile to dissect its function in future work.

In this study, the mtDNA COI genes of 124 individuals in Brachionus calyciflorus complex were sequenced and analyzed. Overall, 84 mtDNA haplotypes were defined, and were split into three clades by the phylogenetic trees. The divergences of COI gene sequence among the three clades ranged from 11.4 to 22.5%, indicating the occurrence of three cryptic species (cryptic species 1, cryptic species 2, and cryptic species 3). Within cryptic species 3, a remarkable degree of differentiation (F st = 0.0947) might be attributable to habitat fragmentation, restricted gene flow, and founder effect, most probably together with local adaptation. The more shared haplotypes were observed, and a non-significant correlation existed between geographic and genetic distance, suggesting that some ancestral haplotypes might be involved in a past range expansion, and as founders give a similar distribution pattern among geographic populations. The networks were also in agreement with the pattern of genetic differentiation in B. calyciflorus species complex revealed by molecular phylogeny. The nested clade analysis suggested a slight geographic structure in the network I, network II, and the highest nesting clade levels within the network III. Under the possible effect of the Younger Dryas Event, the three cryptic species might have survived in multiple relict refugia throughout the south of China. The co-occurrence of cryptic species 2 and 3 might be found in Guangzhou and Hainan due to the possible interference of the barrier and contiguous range expansion. After the Younger Dryas Event, cryptic species 3 with high colonization ability might have arrived and colonized whole habitats at first, and reduce effective gene flow among cryptic species, thus effectively increasing persistence of founder events. Cryptic species 2 might, respectively, have expanded their ranges into Wuhu and Nanjing regions only because of the probable barrier of the Yangtze River, its weak colonization abilities and the 'priority effects' of cryptic species 3. Cryptic species 1 distributed exclusively in Danzhou probably failed to expand its ranges into mainland China because of small population size, weaker dispersal capacity, and the barrier of the Qiongzhou Strait.

Little research has focused on how rotifer communities respond to eutrophication based on their combined taxonomic and functional indices. In this research, the relationship of the environment and rotifer communities was comparatively investigated in two subtropical lakes over one year. The taxon-based indices, including species number (S), Margalef index (D), Simpson index (d), Shannon-wiener index (H′), and functional traits relying on the guild ratio (GR) and the modified guild ratio (GR′) from the moderately eutrophic Lake Xiyanghu were significantly lower than those from the slightly eutrophic Lake Jinghu. Redundancy analysis (RDA) showed that both lakes were distinct from each other. Taken together, the findings indicate that trophic state was an important factor affecting rotifer community structure. In addition, the average annual GR′ of Lake Xiyanghu was <0, suggesting the dominancy of microphagous rotifers. Over time, S, D, d, and H′ were positively correlated with temperature and phosphorus levels in Lake Jinghu, but were negatively correlated with NH4 +-N levels in Lake Xiyanghu. Only GR′ was negatively associated with chlorophyll-a in Lake Xiyanghu, implying that the functional index (GR′) might be an effective tool to explore the relationship between trophic state and the rotifer community in seriously eutrophic lakes.