Mitogenomic phylogeny, biogeography, and cryptic divergence of the genus Silurus (Siluriformes: Siluridae).

The genus Silurus, an important group of catfish, exhibits heterogeneous distribution in Eurasian freshwater systems. This group includes economically important and endangered species, thereby attracting considerable scientific interest. Despite this interest, the lack of a comprehensive phylogenetic framework impedes our understanding of the mechanisms underlying the extensive diversity found within this genus. Herein, we analyzed 89 newly sequenced and 20 previously published mitochondrial genomes (mitogenomes) from 13 morphological species to reconstruct the phylogenetic relationships, biogeographic history, and species diversity of Silurus. Our phylogenetic reconstructions identified eight clades, supported by both maximum-likelihood and Bayesian inference. Sequence-based species delimitation analyses yielded multiple molecular operational taxonomic units (MOTUs) in several taxa, including the Silurus asotus complex (four MOTUs) and Silurus microdorsalis (two MOTUs), suggesting that species diversity is underestimated in the genus. A reconstructed time-calibrated tree of Silurus species provided an age estimate of the most recent common ancestor of approximately 37.61 million years ago (Ma), with divergences among clades within the genus occurring between 11.56 Ma and 29.44 Ma, and divergences among MOTUs within species occurring between 3.71 Ma and 11.56 Ma. Biogeographic reconstructions suggested that the ancestral area for the genus likely encompassed China and the Korean Peninsula, with multiple inferred dispersal events to Europe and Central and Western Asia between 21.78 Ma and 26.67 Ma and to Japan between 2.51 Ma and 18.42 Ma. Key factors such as the Eocene-Oligocene extinction event, onset and intensification of the monsoon system, and glacial cycles associated with sea-level fluctuations have likely played significant roles in shaping the evolutionary history of the genus Silurus.

Whole-genome resequencing confirms the genetic effects of dams on an endangered fish Hemibagrus guttatus (Siluriformes: Bagridae): A case study in a tributary of the Pearl River.

Dam construction in riverine ecosystems has fragmented natural aquatic habitats and has altered environmental conditions. As a result, damming has been demonstrated to threaten aquatic biodiversity by reducing species distribution ranges and hindering gene exchange, leading to the inability to adapt to environmental changes. Knowledge of the contemporary genetic diversity and genetic structure of fish populations that are separated by dams is vital to developing effective conservation strategies, particularly for endangered fish species. We chose the Lianjiang River, a tributary of the Pearl River, as a case study to assess the effects of dams on the genetic diversity and genetic structure of an endangered fish species, Hemibagrus guttatus, using whole-genome resequencing data from 63 fish samples. The results indicated low levels of genetic diversity, high levels of inbreeding and decreasing trend of effective population size in fragmented H. guttatus populations. In addition, there were significant genetic structure and genetic differentiation among populations, suggesting that the dams might have affected H. guttatus populations. Our findings may benefit management and conservation practices for this endangered species that is currently suffering from the effects of dam construction.

Value of post-cerclage transvaginal ultrasound in predicting preterm birth at <28 weeks in twin pregnancy with ultrasound-indicated cerclage.

OBJECTIVE: To evaluate the value of transvaginal ultrasound parameters before and after cerclage in twins in predicting spontaneous preterm birth (sPTB) before 28+0 weeks. METHODS: We retrospectively studied the medical records of twin-pregnant women who underwent ultrasound-indicated cerclage between January 2016 and February 2022 at our hospital. Recorded transvaginal ultrasound images before and after cerclage were reassessed for cervical length (CL), uterocervical angle (UCA), funneling, and sludge. Multivariate logistic and Cox regression analyses were performed to identify the independent risk factors associated with sPTB before 28 weeks. RESULTS: A total of 69 women were included. Among them, 17 women (24.64%) delivered before 28 weeks of age. Regression analysis revealed a significant association of post-cerclage CL, UCA, white blood cell (WBC) count, and gestational age (GA) at cerclage with sPTB before 28 weeks. The area under the curve of these predictors was 0.938 (95% confidence interval, 0.882-0.994; p < .001), with a sensitivity of 88.2%, specificity of 92.3%, positive predictive value of 78.9%, and negative predictive value of 96.0%. Cox analysis showed that post-cerclage UCA was an independent risk factor affecting the cerclage-to-delivery interval (hazard ratios, 1.026; 95% confidence interval (CI), 1.004-1.048; p = .019). CONCLUSIONS: The combination of post-cerclage CL, UCA, WBC count, and GA at cerclage showed good performance in predicting sPTB at <28 weeks in twin pregnancy. Post-cerclage UCA is also associated with pregnancy latency. We found that post-cerclage cervical ultrasound may be useful to predict preterm birth before 28 weeks in twins who undergo ultrasound-indicated cerclage.

Fish Gut Microbiome Analysis Provides Insight into Differences in Physiology and Behavior of Invasive Nile Tilapia and Indigenous Fish in a Large Subtropical River in China.

The gut microbiome is thought to play vital roles in host fitness and local adaptation to new environments, thereby facilitating the invasion of the host species. The Nile tilapia (Oreochromis niloticus) (NT) is an aggressive and omnivorous species that competes with native fishes for food resources, and it has successfully invaded much of the Pearl River basin in China. Here, we investigated the gut microbiomes of invasive Nile tilapia and indigenous black Amur bream (BA) in the same river section using high-throughput 16S rRNA gene sequencing. The results indicated that the gut microbiome of NT had several special characteristics, e.g., higher alpha diversity and greater niche breadth, compared with the bream. The gut microbiota of the small size of Nile tilapia (NTS) and small size of black Amur bream (BAS) groups were dominated by Proteobacteria, while those of the NTS and large size of Nile tilapia (NTL) and BAS and large size of black Amur bream (BAL). BAL and NTL were characterized by Firmicutes and Fusobacteriota, respectively. We found that Pseudomonas, Cetobacterium, Ralstonia, and Romboutsia were biomarkers of the NTS, NTL, BAS, and BAL groups, respectively. Moreover, the results collectively suggested that the clustering coefficients of BAL and NTL networks were greater than those of BAS and NTS networks, and BAS had the smallest network among the four groups. Positive interactions between two ASVs dominated the BAS, NTS, and NTL networks, while the proportion of negative interactions between two ASVs in the BAL network was remarkably increased. Low levels of interspecies competition in the NT gut microbiome would contribute to high diversity in the dietary niches and would also benefit the survival and local adaptation of the host. Our results identified specific biomarkers of gut microbial species in invasive Nile tilapia and provided useful information concerning how to monitor and manage invasive Nile tilapia populations.

Genetic Structure of an East Asian Minnow (Toxabramis houdemeri) in Southern China, with Implications for Conservation.

River dynamics have been hypothesized to substantially influence the genetic structure of freshwater fish taxa. Southern China harbors abundant independent river systems, which have undergone historical rearrangements. This river system is thus an excellent model with which to test the abovementioned hypothesis. In this study, a cyprinid widespread in many independent rivers in southern China, Toxabramis houdemeri, was chosen as an exemplar species with which to explore the effects of river configuration changes on spatial genetic structure using mitochondrial and nuclear markers. The results indicated that the T. houdemeri populations fell into four mitochondrial haplotype groups, each genetically endemic to a single river or two adjacent river systems. The mitochondrial haplotype network recovered a clear genetic boundary between Hainan Island populations and mainland populations. Notable genetic differentiation was observed within populations from distinct river systems in both mitochondrial and nuclear loci. River system separation, mountain barriers, and mobility were the key factors shaping the genetic structure of T. houdemeri populations. Late Pleistocene divergence and historical immigration were identified within the four mitochondrial haplotype groups, indicating that river rearrangements triggered by the Late Pleistocene glacial cycles were important drivers of the complex genetic structure and demographic history of T. houdemeri. Historical demographics suggested that T. houdemeri populations expanded during the Late Pleistocene. The present study has important consequences for the management and conservation of T. houdemeri.

Gut microbiomes of cyprinid fish exhibit host-species symbiosis along gut trait and diet.

Teleost omnivorous fish that coexist partially sharing resources are likely to modify their gut traits and microbiome as a feedback mechanism between ecological processes and evolution. However, we do not understand how the core gut microbiome supports the metabolic capacity of the host and regulates digestive functions in specialized omnivorous fish gut traits. Therefore, we evaluated the gut microbiome of eight omnivorous fish from a single family (i.e., Cyprinidae) in the current study. We examined the correlation between host phylogeny, diet composition, and intestinal morphological traits related to the intestinal microbiome. The results indicated that cyprinid fish with similar relative gut lengths had considerable gut microbiome similarity. Notably, the SL (short relative gut length) group, as zoobenthos and zooplankton specialists, was abundant in Proteobacteria and was less abundant in Firmicutes than in the ML (medium relative gut length) and LL (long relative gut length) groups. These fish could extract nutrients from aquatic plants and algae. Additionally, we found the relative abundance of Clostridium and Romboutsia to be positively correlated with host relative gut length but negatively correlated with the relative abundance of Cetobacterium, Plesiomonas, Bacteroides, and Lactobacillus, and host-relative gut length. We also show a positive linear relationship between host gut microbiome carbohydrate metabolism and relative gut length, while the amino acid and lipid metabolism of the gut microbiome was negatively correlated with host-relative gut length. In addition, omnivorous species competing for resources improve their ecological adaptability through the specialization of gut length, which is closely related to variation in the synergy of the gut microbiome. Above all, specialized gut microbiota and associated gut morphologies enable fish to variably tolerate resource fluctuation and improve the utilization efficiency of nutrient extraction from challenging food resources.

Large-scale DNA barcoding of the subfamily Culterinae (Cypriniformes: Xenocyprididae) in East Asia unveils a geographical scale effect, taxonomic warnings and cryptic diversity.

Geographical scale might be expected to impact significantly the efficiency of DNA barcoding as spatially comprehensive sampling provides opportunities to uncover intricate relationships among closely related species and to detect cryptic diversity for widespread taxa. Here, we present a DNA barcoding study on a Xencyprididae subfamily (Culterinae) involving the production of 998 newly generated DNA barcodes from East Asian drainages (80 localities). Together with 513 barcodes mined from BOLD and GenBank, a reference library consisting of 1511 DNA barcodes (116 localities) for 42 species was assembled, accounting for 66% of known Culterinae species. Intraspecific genetic distances are positively correlated to geographical scale, while a negative correlation is detected between interspecific genetic distances and geographical scale. The present study demonstrates that geographical scale influences the efficiency of DNA barcoding by narrowing the width of the barcoding gap. DNA-based species delimitation analyses delimited 44 molecular operational taxonomic units (MOTUs). Rampant cryptic diversity is detected within eight species with multiple MOTUs, whereas 25 species present mismatch between morphological and molecular delimitations. A total of 18 species are lumped into nine MOTUs due to low interspecific divergence and/or mixed lineages. Several MOTU divergences are hypothesized to relate to known biogeographical barriers and geological events during the Pliocene and Pleistocene. This study provides new insights into the taxonomy and phylogeography of the subfamily Culterinae.

Clinical Observation of Effects of Prepregnancy Body Mass Index and Weight Gain during Pregnancy on Neonatal Weight and Delivery Outcome.

Objective. The increase of BMI before pregnancy and during pregnancy will lead to hypertensive disorder in pregnancy (HDP) and abnormal glycolipid metabolism, as well as increase the risk of neonatal weight abnormalities and adverse pregnancy outcome. This study retrospectively analyzed the clinical data of 358 women who were admitted to the obstetrics department of our hospital from January 2018 to July 2019. And the relationship between prepregnancy BMI, BMI increase during pregnancy and delivery methods, postpartum hemorrhage, neonatal weight, premature delivery, neonatal asphyxia, gestational diabetes mellitus (GDM), and hypertension during pregnancy (HDP) was observed and compared. The results show that both high BMI before pregnancy and excessive weight gain during pregnancy can lead to the incidence of abnormal birth weight and adverse birth outcome increase. Therefore, in order to reduce the incidence of neonatal weight abnormalities and adverse birth outcomes and to prevent the adverse effects during pregnancy and postpartum, clinically, obese women should be guided to eat according to reasonable diet and exercise to control their weight.

DNA barcoding reveals the temporal community composition of drifting fish eggs in the lower Hongshui River, China.

Determining the temporal community composition of fish eggs in particular regions and understanding the reproductive times of regional fish taxa are key aspects of the management and regulation of regional fish stocks. However, it is extremely difficult to accurately identify fish eggs due to the absence of diagnostic morphological characters. We sampled fish eggs in the lower Hongshuihe River (an upper mainstem of the Pearl River) between May and September 2020. We then used DNA barcoding to determine the species composition of the egg pool and to predict the spawning periods of the identified species. A total of 641 eggs and 17 larvae were chosen for molecular identification; 397 eggs and 17 larvae yielded high-quality barcoding sequences. The high failure rate (~38%) was most likely due to long-term storage in low concentrations of ethanol prior to molecular analysis. We successfully classified 392 eggs into 10 species and 13 larvae into four species using public databases. Most of the species identified in the egg pool were small and/or benthic, and migratory species were rare. This may partially reflect the adverse effects of hydropower cascade development in this river section. We also found that spawning periods tended to be species-specific. Our study provides a reference for the conservation and management of regional fishery stocks.

Determination of the complete mitochondrial genome of Pseudobagrus gracilis (Siluriformes: Bagridae) and its phylogeny.

Pseudobagrus gracilis is an endemic bagrid catfish in the Pearl River. To date, sparse studies conducted on this species have blocked our understanding of this species. In this study, the complete mitochondrial genome of P. gracilis was sequenced and reported using Illumina MiSeq platform. The P. gracilis mitogenome was 16,527 bp in length and comprised 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes, and one control region (D-loop). Its overall nucleotide base composition was 31.0% (A), 15.6% (G), 26.3% (C), and 27.1% (T), with an AT content 58.1%. Phylogenetic analyses based on Neighbor-joining approach revealed that Pseudobagrus species formed three lineages (I, II and III) and P. gracilis had close relationship with P. emarginatus, P. pratti and P. truncatus.

Sequencing an F1 hybrid of Silurus asotus and S. meridionalis enabled the assembly of high-quality parental genomes.

Genome complexity such as heterozygosity may heavily influence its de novo assembly. Sequencing somatic cells of the F1 hybrids harboring two sets of genetic materials from both of the paternal and maternal species may avoid alleles discrimination during assembly. However, the feasibility of this strategy needs further assessments. We sequenced and assembled the genome of an F1 hybrid between Silurus asotus and S. meridionalis using the SequelII platform and Hi-C scaffolding technologies. More than 300 Gb raw data were generated, and the final assembly obtained 2344 scaffolds composed of 3017 contigs. The N50 length of scaffolds and contigs was 28.55 Mb and 7.49 Mb, respectively. Based on the mapping results of short reads generated for the paternal and maternal species, each of the 29 chromosomes originating from S. asotus and S. meridionalis was recognized. We recovered nearly 94% and 96% of the total length of S. asotus and S. meridionalis. BUSCO assessments and mapping analyses suggested that both genomes had high completeness and accuracy. Further analyses demonstrated the high collinearity between S. asotus, S. meridionalis, and the related Pelteobagrus fulvidraco. Comparison of the two genomes with that assembled only using the short reads from non-hybrid parental species detected a small portion of sequences that may be incorrectly assigned to the different species. We supposed that at least part of these situations may have resulted from mitotic recombination. The strategy of sequencing the F1 hybrid genome can recover the vast majority of the parental genomes and may improve the assembly of complex genomes.

Temporal species-level composition of larvae resources in the lower Pearl River drainage and implications for species' reproductive cycles.

Resolving the temporal community composition of a larvae population can not only further our understanding of the regional species composition but also help us to infer the reproductive times of regional fish taxa, which can have implications on the development of effective monitoring and conservation policies for the regional fish stock. Nevertheless, correctly diagnosing the fish larvae is extremely challenging due to the paucity of diagnostic morphological characters at the species level. Based on daily larval samplings during March and October in 2018, this study combined morphological features with a DNA barcode technique to determine the species composition of fish larvae in the lower Pearl River drainage (LPR) and evaluate the spawning periods of identified species. Due to an absence of reference barcodes for LPR fishes, a DNA barcode library of adult fishes in the LPR was built for 384 individuals representing 78 morphological species. Analyses demonstrated the usability of the barcode library and uncovered many undetected mitochondrial lineages in 12 species. Morphological analyses performed on 81 temporal larval samples revealed 25 morphotypes and assigned 9 morphotypes into the species level. A total of 1624 larvae from 96 temporal larval samples were selected for molecular identification, and high quality barcoding sequences were obtained from 1391 larvae. We accurately assigned 1078 larvae to 37 species using our barcode library and published database. Among the identified species, a critically endangered species, namely, Ochetobius elongatus, and several invasive species were examined, providing a new perspective to assess the stock of regional endangered and invasive species. Furthermore, this study found high species diversity occurred primarily between May and September, and clarified the spawning periods of identified species inferred from the temporal occurrences of larvae. Above all, our study highlights the applicability to fish larval ecology to assist conservation and fishery management efforts.

Individual dietary specialization reduces intraspecific competition, rather than feeding activity, in black amur bream (Megalobrama terminalis).

Individual specialization and high plasticity in feeding activity are common in natural populations. However, the role of these two in intraspecific competition is unclear. In this study, the rhythm of feeding activity, dietary composition, niche width, niche overlap, and individual specialization was explored in four different size groups of black amur bream (Megalobrama terminalis), using microscopic identification of foregut contents and stable isotope analysis (δ13C and δ15N) of dorsal muscle. Both methods observed ontogenetic shifts in dietary preference and individual specializations, and revealed that the total niche width of large individuals was greater than small individuals. Mixed linear models indicated that feeding activity was significantly influenced by time (p < 0.0001), and no significant changes among size groups was evident (p = 0.244). Niche overlaps revealed that there was intensive diet competition between different size groups of black amur bream. Individual specialization in small juveniles was likely to be stronger than sub-adult and adult groups. Pearson's correlation analysis revealed that the individual specialization was positively correlated with mean diet similarity within a group. The results indicated that intraspecific competition is reduced mainly by individual dietary specialization, rather than shift in feeding activity.

Insight into the combinatorial transcriptional regulation on α-amylase gene in animal groups with different dietary nutrient content.

Gene expression is generally regulated by multiple transcription factors (TFs). Despite previous findings of individual TFs regulating pancreatic α-amylase gene expression, the combinatorial transcriptional regulation is not fully understood. To gain insight into multiple TF regulation for pancreatic α-amylase gene, we employed a function conservation approach to predict interacting TFs regulating pancreatic α-amylase gene for 3 dietary animal groups. To this end, we have identified 77, 25, and 118 interacting TFs for herbivore, omnivore, and carnivore, respectively. Computational modeling of TF regulatory networks demonstrated that known pancreas-specific TFs (e.g. GR, NFAT, and PR) may play important roles in recruiting non pancreas-specific TFs to the TF-TF interaction networks, offering specificity and flexibility for controlling pancreatic α-amylase gene expression in different dietary animal groups. The findings from this study indicate that combinatorial transcriptional regulation could be a critical component controlling pancreatic α-amylase gene expression.

Complete mitochondrial genome of the freshwater fish, Carassioides acuminatus (Cypriniformes, cyprinidae).

In this study, the complete mitochondrial genome of Carassioides acuminatus was first sequenced and annotated. The entire mitogenome is 16,579bp in length, which consists of 13 protein-coding genes (PCGs), 22 transfer RNA genes, 2 ribosomal RNA genes, and a control region (D-loop). The overall nucleotide composition of the C. acuminatus mitochondrial genome shows an obvious anti-G bias. The accuracy of the fresh sequences was verified by phylogenetic analysis. The complete mitochondrial genome of C. acuminatus is useful to population genetics and molecular systematics.

Placenta growth factor promotes migration through regulating epithelial-mesenchymal transition-related protein expression in cervical cancer.

Epithelial-mesenchymal transition (EMT) plays an important role in cancer invasion and metastasis by enabling cancer cells to depart from the primary tumor, invade surrounding tissue and disseminate to distant organs. The existence and function of EMT in cervical cancer is poorly understood. Placental growth factor (PLGF) has been shown to associate with EMT in various cancers. However, whether PLGF is involved in EMT in cervical cancer remains unclear. Thus the present study examined the relationship between PLGF expression and EMT-related proteins in 110 cervical lesions samples. We detected that PLGF was expressed in 61.8% cervical lesion sections. In addition, PLGF expression is positively correlated with low expression level of E-cadherin and high expression level of vimentin. Serum samples and cervical lavage samples were collected from patients with pre-invasive and invasive lesion of uterine cervix or normal control group, the PLGF levels were determined by enzyme-linked immunosorbent assay (ELISA). We found that a significantly high level of PLGF could be detected both in serum and vaginal lavage compared with normal women group, and there is no significant difference between serum and lavage in PLGF level. In addition, whatever in lavage or in serum, the PLGF level in stage I and II was significantly higher than it in CINIII or cancer in situ. However, there is no significant difference between the stage I and stage II; we also found that exogenous PLGF promotes molecular changes of epithelial-mesenchymal transition (EMT) in siha cells. In addition, application of a specific EKR1/2 inhibitor could reverse the effects of PLGF. These findings suggested that PLGF could regulate the expression of EMT-related proteins and promote migration of siha cells through ERK/MAPK signaling pathway. Therapies that targets PLGF/Flt-1/ERK/MAPK signaling pathway may be beneficial in treatment of cervical cancer.