Exploring the underlying mechanism by transcriptome sequencing in rats with high-voltage electrical burns and the role of iron metabolism.

BACKGROUND: Clinically, the condition of skeletal muscle injury is the key to the process of high voltage electrical burn (HVEB) wound repair. The aim of this study was to identify the potential mechanisms and intervention targets of skeletal muscle injury after HVEB. METHODS: A skeletal muscle injury model in SD rats with HVEB was made. Pathological examination and transcriptome sequencing of injured skeletal muscles were performed, and the expression levels of key proteins and genes in related signaling pathways were verified. RESULTS: Skeletal muscle injury was progressively aggravated within 48 h, then the injury was gradually repaired with scar formation occurring within 1 week. The mechanism of skeletal muscle injury is complex and varied, and ferroptosis is one of the mechanisms. The ferrous iron content in the injured skeletal muscle tissue of model rats increased significantly at 24 h after injury. After 24 h, damage to injured skeletal muscle tissue could be alleviated by increasing iron storage and blocking lysosomal phagocytosis of autophagy. CONCLUSIONS: Skeletal muscle injury caused by HVEB is characterized by adjacent endangered tissue progression after injury. Ferroptosis is involved in the mechanism of HVEB, and iron metabolism-related proteins may be potential targets for preventing progressive skeletal muscle injury.

A spider mating plug functions to protect sperm.

Mating plugs in animals are ubiquitous and are commonly interpreted to be products of mating strategies. In spiders, however, mating plugs may also take on functions beyond female remating prevention. Due to the vagaries of female genital (spermathecal) anatomy, most spiders face the problem of having to secure additional, non-anatomical, protection for transferred sperm. Here, we test the hypothesis that mating plugs, rather than (or in addition to) being adaptations for mating strategies, may serve as sperm protection mechanism. Based on a comparative study on 411 epigyna sampled from 36 families, 187 genera, 330 species of entelegyne spiders, our results confirm the necessity of a sperm protection mechanism. We divided the entelegyne spermathecae into four types: SEG, SED, SCG and SCD. We also studied detailed morphology of epigynal tracts in the spider Diphya wulingensis having the SEG type spermathecae, using 3D-reconstruction based on semi thin histological series section. In this species, we hypothesize that two distinct types of mating plug, the sperm plug and the secretion plug, serve different functions. Morphological details support this: sperm plugs are formed on a modified spermathecal wall by the spilled sperm, and function as a temporary protection mechanism to prevent sperm from leaking and desiccating, while secretion plugs function in postcopulation both as a permanent protection mechanism, and to prevent additional mating. Furthermore, with the modified spermathecal wall of S2 stalk, the problem of shunt of sperm input and output, and the possibility of female multiple mating have been resolved. Variation in spermathecal morphology also suggests that the problem of sperm protection might be resolved in different ways in spiders. Considering mating plugs of varying shapes and origins in the vast morphospace of spiders, we conclude that mating plugs might serve different purposes that relate both to mating strategies, as well as to sperm protection.

Oxidative stress mediates hippocampal neuronal apoptosis through ROS/JNK/P53 pathway in rats with PTSD triggered by high-voltage electrical burn.

BACKGROUND: The pathogenesis of post-traumatic stress disorder (PTSD) triggered by high-voltage electrical burn (HVEB) remains unclear and the oxidative stress plays a role in this process. The purpose of this study is to investigate the underlying mechanism of oxidative stress mediates hippocampal neuronal apoptosis in rats with PTSD triggered by HVEB. MATERIALS AND METHODS: The PTSD rat model was developed by stimulating with high voltage electricity and screened using behavioral performance including Morris water maze (MWM), elevated plus-maze (EPM) and open-field test (OFT). The reactive oxygen species (ROS) generation was measured by DHE fluorescence staining or flow cytometry. Western blotting assay was used to detect the proteins of p-JNK, JNK, P53, PUMA, Bcl-2 and Bax in hippocampal tissue or HT22 cells treated with electrical stimulation. RESULTS: The serum MDA and 8-OHdG levels were increased (P < 0.001), while the activities of SOD and CAT were decreased (P < 0.001) significantly in patients with HVEB. Behavioral test results showed that high-voltage electric stimulation induced the PTSD-like symptoms and the ROS-JNK-P53 pathway was involved in the neuronal apoptosis in rats with PTSD induced by HVEB. In vitro experiments further confirmed the electrical stimulation induced neuronal apoptosis through ROS/JNK/P53 signaling pathway and the antioxidant NAC could rescued the ROS generation, activation of JNK/P53 proteins and improved the cell apoptosis rate in HT22 cells. Finally, the JNK inhibitor SP600125 could significantly inhibited the percentage of HT22 cell apoptosis induced by electrical stimulation (P < 0.001). CONCLUSIONS: These results indicated that oxidative stress mediates hippocampal neuronal apoptosis through ROS/JNK/P53 pathway in rats with PTSD triggered by HVEB.

Erythromycin resistance of clinical Campylobacter jejuni and Campylobacter coli in Shanghai, China.

Campylobacter species are zoonotic pathogens, as well as the prevalent cause of foodborne bacterial gastroenteritis. The spread of antimicrobial-resistant strains poses a serious threat to global public health and attracts attention worldwide, but information about clinical Campylobacter is relatively limited compared to isolates from food and animals. The current study illustrated the prevalence and antimicrobial resistance profiles of Campylobacter jejuni and Campylobacter coli isolates collected from a consecutive surveillance program between 2012 and 2019 in Shanghai, China, using antimicrobial susceptibility testing and whole-genome sequencing. Among the 891 Campylobacter strains (761 C. jejuni and 130 C. coli) isolates collected, high portions above 90% of resistance to ciprofloxacin, nalidixic acid, and tetracycline were observed for both C. jejuni and C. coli. The most common MDR profiles represented by C. jejuni and C. coli were combination of ciprofloxacin, tetracycline, florfenicol and nalidixic acid (5.39%), and azithromycin, ciprofloxacin, erythromycin, gentamicin, tetracycline, clindamycin, nalidixic acid (28.46%), respectively. The erythromycin resistance of C. coli (59.23%) is higher than C. jejuni (2.50%). A total of 76 erythromycin resistant isolates (16 C. jejuni and 60 C. coli) were sequenced using Illumina platform for determining the genotypes, antimicrobial resistance patterns and phylogeny analysis. Multilocus sequence typing (MLST) analysis showed a high genetic diversity with 47 sequence types (STs), including 4 novel alleles and 12 new STs. The most abundant clonal complexes (CCs) were CC-403 (31.25%) and CC-828 (88.33%) for C. jejuni and C. coli, respectively. Among the 76 erythromycin-resistant isolates, mutation A2075G in 23S rRNA and erm(B) gene were detected in 53.95 and 39.47%, respectively. The erm(B) gene was identified exclusively in 30 C. coli isolates. All these erm(B) positive isolates were multi-drug resistant. Furthermore, comparison of the erm(B)-carrying isolates of multiple sources worldwide demonstrated the possibility of zoonotic transmission of erm(B) in Campylobacter. These findings highlight the importance of continuous surveillance of erythromycin resistance dissemination in Campylobacter which may compromise the effectiveness of antimicrobial therapy.

Changing Molecular Epidemiology of Vibrio cholerae Outbreaks in Shanghai, China.

The 7th cholera pandemic began in 1961 in Sulawesi, Indonesia, and then spread around the world in at least three waves. However, the lack of genome sequences for Vibrio cholerae strains under long-term surveillance in East Asia, especially in China, has restricted our understanding of the dynamics of the intracountry and intercountry evolution and transmission of the 7th-pandemic clones. In this study, we obtained the genome sequences of 60 V. cholerae strains isolated in Shanghai, the largest port in the world and the largest city in China, from 1961 to 2011. Our whole-genome-based phylogeny of 7th-pandemic strains revealed that all but one fell into five "stages," most of which are single clades and share independent ancestors. Each stage dominated in succession for a period, with little overlap between them. In addition, two near-identical Shanghai strains belonging to a pre-7th-pandemic precursor and 4 nontoxigenic O1/O139 strains attributed to independent recombination events at the O-antigen loci were present. The major lineages of the 7th pandemic in Shanghai appeared to be closely related to V. cholerae strains isolated from South or Southeast Asia. Stage succession was consistently related to changes in society and human activity, implying that human-caused niche change may play a vital role in the cholera dynamics in Shanghai.IMPORTANCE V. cholerae is the causative agent of cholera, a life-threatening disease characterized by severe, watery diarrhea. The 7th pandemic started in Indonesia in 1961 and spread globally, currently infecting 1.3 million to 4 million people annually. Here, we applied whole-genome sequencing to analyze a long-term collection of V. cholerae clinical strains to reveal the phylogenetic background and evolutionary dynamics of the 7th pandemic in Shanghai, which had undergone breathtakingly rapid development in the last half-century. All but one of the Shanghai 7th-pandemic strains fell into five "stages" that were dominant in Shanghai and appeared to be closely related to 7th-pandemic strains of South or Southeast Asia. Our findings extended the understanding of the dynamics of the evolution and transmission of the 7th-pandemic clones in East Asia and the relationship between social changes and cholera epidemiology.

Comparative morphology refines the conventional model of spider reproduction.

Our understanding of spider reproductive biology is hampered by the vast anatomical diversity and difficulties associated with its study. Although authors agree on the two general types of female spider genitalia, haplogyne (plesiomorphic) and entelegyne (apomorphic), our understanding of variation within each group mostly concerns the external genital part, while the internal connections with the reproductive duct are largely unknown. Conventionally and simplistically, the spermathecae of haplogynes have simple two-way ducts, and those of entelegynes have separate copulatory and fertilization ducts for sperm to be transferred in and out of spermathecae, respectively. Sperm is discharged from the spermathecae directly into the uterus externus (a distal extension of the oviduct), which, commonly thought as homologous in both groups, is the purported location of internal fertilization in spiders. However, the structural evolution from haplo- to entelegyny remains unresolved, and thus the precise fertilization site in entelegynes is ambiguous. We aim to clarify this anatomical problem through a widely comparative morphological study of internal female genital system in entelegynes. Our survey of 147 epigyna (121 examined species in 97 genera, 34 families) surprisingly finds no direct connection between the fertilization ducts and the uterus externus, which, based on the homology with basal-most spider lineages, is a dead-end caecum in entelegynes. Instead, fertilization ducts usually connect with a secondary uterus externus, a novel feature taking over the functional role of the plesiomorphic uterus externus. We hypothesize that the transition from haplo- to entelegyny entailed not only the emergence of the two separate duct systems (copulatory, fertilization), but also involved substantial morphological changes in the distal part of the oviduct. Thus, the common oviduct may have shifted its distal connection from the uterus externus to the secondary uterus externus, perhaps facilitating discharge of larger eggs. Our findings suggest that the conventional model of entelegyne reproduction needs redefinition.

Fluoroquinolone Resistance Mechanisms in Shigella Isolates in Shanghai, China, Between 2010 and 2015.

OBJECTIVES: This study aimed to investigate the antimicrobial susceptibility of Shigella isolated in Shanghai, China and to determine the genetic basis of its resistance to fluoroquinolones. MATERIALS AND METHODS: A total of 402 strains of Shigella, including 145 Shigella flexneri and 257 Shigella sonnei isolates, were analyzed. The Kirby-Bauer disk diffusion method was used to determine the susceptibility of the strains to 13 antimicrobials. Minimum inhibitory concentration of ciprofloxacin was determined by E-test. Mutations within the quinolone resistance-determining regions (QRDRs) of gyrA and parC and in the plasmid-mediated quinolone resistance (PMQR) genes, including qnrA, qnrB, qnrS, and aac (6')-Ib-cr, were detected by polymerase chain reaction. All the products were then sequenced. RESULTS: Most of the Shigella isolates were found to be resistant to nalidixic acid (96.4%), streptomycin (96.4%), ampicillin (86.2%), tetracycline (79.8%), and sulfamethoxazole/trimethoprim (80.6%). S. flexneri isolates showed a significantly higher resistance to cefepime (33.6%), ciprofloxacin (54.2%), norfloxacin (34.1%), and levofloxacin (12.1%) compared with that observed for the S. sonnei strains (χ2 analysis, p < 0.05). Three mutations (Ser83, Asp87, and His211) in gyrA and one mutation (Ser80) in parC were detected. Of 257 S. sonnei isolates, 11.7% possessed gyrA mutations and 2% had parC mutations. Of 145 S. flexneri isolates, 98.6% possessed gyrA mutations and 97.9% had parC mutations. The plasmid-mediated resistance genes of qnrS and aac (6')-Ib-cr were detected among 17 strains (4.2%). CONCLUSIONS: The mutation percentage within the QRDR of S. flexneri was as high as 98.6 in gyrA and 97.9 in parC. The significant abundance of mutations within QRDRs conferred high levels of fluoroquinolone resistance. Moreover, the PMQR genes, particularly qnrS, played an important role in the decreased susceptibility of Shigella to fluoroquinolones.

A review of the Arcuphantes species with the genitalia of arcuatulus-type Araneae, Linyphiidae).

This is the second study addressing the Nearctic Arcuphantes species. Re-examination of Arcuphantes deposited in the American Museum of Natural History shows that four species have the genitalia of arcuatulus-type, including three new species: Arcuphantes arcuatulus (Roewer, 1942), A. cruciatus n. sp., A. denticulatus n. sp. and A. semiorbiculatus n. sp. Genital characters of arcuatulus-type are reviewed and compared with those of fragilis-type based on SEM images and microscopic pictures. Descriptions of the new species and a redescription of the known species are provided.

A new species of the spider genus Solenysa from China (Araneae, Linyphiidae).

The genus Solenysa Simon, 1894 belongs to Linyphiidae Blackwall, 1859, which is a species-rich group, including 608 genera and 4,571 species (World Spider Catalog 2018). Solenysa currently includes 14 species from China, Japan and the Korean Peninsula (Simon 1894; Namkung 1986; Li Song 1992; Gao, Zhu Sha 1993; Tu, Ono Li 2007; Ono 2011; Tu Hormiga 2011; Wang, Ono Tu 2015). The linyphiid phylogeny based on molecular data shows that Solenysa species forms one of the seven main clades within Linyphiidae (Wang et al. 2015). According to the phylogenetic analysis based on morphological data, Tu and Hormiga (2011) divided the genus Solenysa into four species groups, each having a unique genital type comprised by series genital characters. As an old branch with a long evolutionary history, Solenysa spiders have accumulated a long list of synapomorphies (Tu Hormiga 2011), not only having a unique somatic appearance, but also specific genitalic characters that distinguish them from all other linyphiids.

An emm5 Group A Streptococcal Outbreak Among Workers in a Factory Manufacturing Telephone Accessories.

Ranked among the top10 infectious causes of death worldwide, group A Streptococcus (GAS) causes small- and large-scale outbreaks, depending on the trigger as transmission of a GAS strain or expansion of predominant clones. In China, GAS infections other than scarlet fever are not notifiable. In Shanghai, an epidemiological investigation was initiated after two successive severe pneumonia cases with one death in a digital factory, from where outbreaks are less widely reported. The investigation was performed using emm typing, pulsed-field gel electrophoresis (PFGE) typing, superantigen profiling, and genome analysis. This enabled characterization of relatedness among the outbreak isolates and identification of the mobile genetic elements present. Among 57 patients with respiratory symptoms investigated in the factory, emm5 GAS strains were isolated from 8 patients. The eight GAS infection cases comprising one fatal severe pneumonia case, six influenza-like illness cases, and one pharyngitis case. Two risk factors were identified: adult with an age of 18-20 years and close contact with a GAS patient or carrier. GAS attack rate was 14.0% (8/57), and GAS carriage rate was probably around 2.7% (14/521) based on surveys in two nearby districts. All the 10 outbreak associated isolates were assigned to emm5 and sequence type ST-99 (emm5/ST-99), harbored superantigen genes speC, speG, and smeZ, and were assigned to two similar PFGE patterns (clones). Among the outbreak associated isolates, all carried ermA with resistance to erythromycin and inducible resistance to clindamycin, and eight (80%) carried a tetM gene with resistance to tetracycline. Among the 14 carriage isolates, 12 were emm12/ST-36, and 2 were emm1/ST-28, all with superantigen genes speC, speG, ssa, and smeZ. All the carriage isolates harbored ermB and tetM with resistance to erythromycin, clindamycin, and tetracycline. Genome analysis showed the two outbreak clones were closely related and possessed new prophages carrying virulence gene sdc and antibiotic resistance genes of ermA and tetM, which were not found in the emm5 reference strain Manfredo. This is the first report of a GAS outbreak in this type of workplace. The outbreak was caused by two closely related emm5 clones that differed from the predominant emm types circulating in China.

On a desmitracheate "micronetine" Nippononeta alpina (Li & Zhu, 1993), comb. n. (Araneae, Linyphiidae).

The phylogenetic analyses based on molecular data demonstrate that all "micronetine" species of a desmitracheate system form a monophyly. Macrargus Dahl, 1886 is a "micronetine" genus, the species of which have a haplotracheate system in general, while Macrargus alpinus Li & Zhu, 1993 was found to have a desmitracheate system; this makes its generic placement problematic. According to the results of phylogenetic analysis, we transfer Macrargus alpinus to another genus as Nippononeta alpina (Li & Zhu, 1993), comb. n., and provide a redescription of its genital characters and somatic features. Comparisons with other "micronetine" species with a desmitracheate system are provided. Putative synapomorphies for Nippononeta, the clade Nippononeta + Agyneta, and for the "desmitracheate micronetines" clade, as well as their relationship with Helophora, are provided and discussed.

Genetic and serological identification of three Vibrio parahaemolyticus strains as candidates for novel provisional O serotypes.

Vibrio parahaemolyticus is a Gram-negative, halophilic Vibrio that naturally inhabits marine and estuarine environments worldwide and has recently been recognized as one of the most important foodborne pathogens. To date, 13 O serotypes and 71 K serotypes of V. parahaemolyticus have been identified. However, untypeable V. parahaemolyticus strains are frequently found during routine detection, indicating that other forms of serotypes exist and suggesting the necessity for extension of the antigenic scheme. In this work, through the genetic analysis of the O serotype genetic determinants (OGDs) and the production of antisera and serological tests, we identified three novel O serotypes of V. parahaemolyticus. Further analyses showed that recombination and gene-set deletions/insertions within OGDs may play key roles in the generation of V. parahaemolyticus O serotype diversity. A PCR method was developed for the identification of these novel O serotypes, and specificity and sensitivity were evaluated. A double-blind test including 283 clinical isolates was performed, giving perfect correlation with the agglutination test results. Generally, our study expanded the O-antigenic scheme of V. parahaemolyticus from 13 to 16 and provided a tool with the potential for the detection and identification of V. parahaemolyticus strains (especially untypeable strains) isolated from both the clinic and the environment.

A review of the Nearctic linyphiid spider Arcuphantes fragilis (Araneae, Linyphiidae) and closely related species.

The Western Nearctic species Arcuphantes fragilis Chamberlin & Ivie, 1943 and several closely related species are reviewed. Seven species are recognized, including two new species: A. cavaticus Chamberlin & Ivie, 1943, A. decoratus Chamberlin & Ivie, 1943, A. fragilis Chamberlin & Ivie, 1943, A. dentatus n. sp., A. curvomarginatus n. sp., A. potteri Chamberlin & Ivie, 1943, and A. sylvaticus Chamberlin & Ivie, 1943. The female of A. decoratus is described for the first time. These seven species share a same genital type, distinguishing them from other congeners, with small differences among them. Illustrations for A. dentatus n. sp., SEM images and digital photographs for all seven species are presented. Descriptions of new species, redescriptions of known species and a key for the seven species are provided.

Resolving the phylogeny of a speciose spider group, the family Linyphiidae (Araneae).

For high-level molecular phylogenies, a comprehensive sampling design is a key factor for not only improving inferential accuracy, but also for maximizing the explanatory power of the resulting phylogeny. Two standing problems in molecular phylogenies are the unstable placements of some deep and long branches, and the phylogenetic relationships shown by robust supported clades conflict with recognized knowledge. Empirical and theoretical studies suggest that increasing taxon sampling is expected to ameliorate, if not resolve, both problems; however, sometimes neither the current taxonomic system nor the established phylogeny can provide sufficient information to guide additional sampling design. We examined the phylogeny of the spider family Linyphiidae, and selected ingroup species based on epigynal morphology, which can be reconstructed in a phylogenetic context. Our analyses resulted in seven robustly supported clades within linyphiids. The placements of four deep and long branches are sensitive to variations in both outgroup and ingroup sampling, suggesting the possibility of long branch attraction artifacts. Results of ancestral state reconstruction indicate that successive state transformations of the epigynal plate are associated with early cladogenetic events in linyphiid diversification. Representatives of different subfamilies were mixed together within well supported clades and examination revealed that their defining characters, as per traditional taxonomy, are homoplastic. Furthermore, our results demonstrated that increasing taxon sampling produced a more informative framework, which in turn helps to study character evolution and interpret the relationships among linyphiid lineages. Additional defining characters are needed to revise the linyphiid taxonomic system based on our phylogenetic hypothesis.

On the desmitracheate "micronetine" genus Nippononeta Eskov, 1992 (Araneae, Linyphiidae).

The desmitracheate system in a "micronetine" genus Nippononeta Eskov, 1992 is recognized for the first time in the present study. This makes the subfamilial placement of this genus problematic. A morphological study was conducted for Nippononetakurilensis Eskov, 1992 (the type species of Nippononeta) and Nippononetacoreana (Paik, 1991). Characters of genitalia and tracheal system, as well as some somatic characters were studied in detail by using scanning electronic microscopy (SEM), and compared with those of Agyneta. Updated descriptions of the genus Nippononeta and its two species are presented. Putative synapomorphies for Nippononeta and Agyneta are provided, as well as some putative synapomorphies shared by the two genera. The results imply that both scaped epigynum and desmitracheate tracheal system are probably homoplastic. The placement of Nippononeta and Agyneta within Linyphiidae need to be resolved in future studies.

A review of Solenysa spiders from Japan (Araneae, Linyphiidae), with a comment on the type species S.mellotteei Simon, 1894.

The present paper gives a review of Solenysa species from Japan and provides a solution for the species bearing the generotype name Solenysamellotteei Simon, 1894. A total of six species are recorded, including two new species Solenysamacrodonta sp. n. and Solenysatrunciformis sp. n. The species collected from Kawasaki (NSMT-Ar 11154) and Hachioji should be the generotype Solenysamellotteei, with Solenysaakihisai Tu, 2011, syn. n. as its junior synonym. To distinguish these congeneric species from each other, their genital characters are provided in detail based on images collected by scanning electron microscopy and light microscopy.

Acanoides gen. n., a new spider genus from China with a note on the taxonomic status of Acanthoneta Eskov & Marusik, 1992 (Araneae, Linyphiidae, Micronetinae).

A new "micronetine" genus Acanoides gen. n. is erected to accommodate two species from China: Acanoides beijingensis sp. n. as the type species and Acanoides hengshanensis (Chen & Yin, 2000), comb. n., with the females described for the first time. The genitalic characters and somatic features of the new genus were studied by means of light microscopy and scanning electron microscopy (SEM). The monophyly of the new genus was tested by a phylogenetic analysis based on molecular data. Descriptions of the new genus, the new species and the new combination are presented; SEM images and microscopy pictures of somatic and genitalic characters are provided in detail. To distinguish from other genera with similar genitalic characters, we compare the new genus with the species of Acanthoneta Eskov & Marusik, 1992, Epibellowia Tanasevitch, 1996 and Wubanoides Eskov, 1986. Four putative synapomorphies for Acanoides gen. n. are suggested to support its monophyly that could be tested in the future. Furthermore, redescriptions of the epigynal morphology of Acanthoneta aggressa Chamberlin & Ivie, 1943 (Nearctic) and on the male of A. dokutchaevi Eskov & Marusik, 1993 (Far East Asia, firstly recorded from China) are provided. Based on comparison with Poeciloneta, from which Acanthoneta stat. n. was separated by Saaristo and Tanasevitch (1996), a revised diagnosis is proposed to support the generic status.

[Characterization of phenotype and molecular characteristics on Vibrio cholerae strains isolated in Shanghai, 1962-2011].

OBJECTIVE: To describe the phenotype and molecular characteristics of Vibrio (V.) cholerae strains isolated in Shanghai, from 1962 to 2011. METHODS: K-B test was used to investigate the antibiotic resistance of V. cholerae strains. PCR was applied to detect seven virulence-related genes including cholera toxin (ctxA), zonula occludens toxin (zot), accessory cholera enterotoxin (ace), hemolysin (hlyA), toxin-coregulated pilus (tcpA) outer membrane protein (ompU) and the regulatory protein genes (toxR). Genetic relation was assessed by pulsed-field gel electrophoresis (PFGE) and the patterns were clustered by BioNumerics software. RESULTS: V. cholerae strains isolated from 1962 to 1996 were sensitive to most of the antibiotics. However, the strains isolated from 2005 to 2011 were resistant to many antibiotics. V. cholerae O139 group showed higher prevalence of resistance to several antibiotics compared with O1 group, and the resistance rate of the O139 toxigenic isolates was higher than that of the non-toxigenic isolates. Most of the O1 strains isolated from 2005 to 2011 were non-toxigenic while O139 strains isolated from 2005 to 2011 were almost toxigenic. There were no strains of ctxA+ detected from the rivers from 2005 to 2011. Main gene type of the O1 strains detected from the aquatic products was hlyA+ toxR+ ompU+, while that of the O139 strains was hlyA+ toxR+ ompU+ ctxA+ ace+ zot+ tcpA+. Using PFGE, 222 V. cholerae strains were subtyped into 121 molecular types. O139 strains were divided to three clusters and O1 strains to five clusters. CONCLUSION: The characteristics of V. cholerae strains isolated in Shanghai from 1962 to 2011 showed great changes, suggesting that more attention should be paid to the multiplication on antibiotic resistance of V. cholerae strains.

Rampant historical mitochondrial genome introgression between two species of green pond frogs, Pelophylax nigromaculatus and P. plancyi.

BACKGROUND: Mitochondrial introgression may result in the mitochondrial genome of one species being replaced by that of another species without leaving any trace of past hybridization in its nuclear genome. Such introgression can confuse the species genealogy estimates and lead to absurd inferences of species history. We used a phylogenetic approach to explore the potential mitochondrial genome introgression event(s) between two closely related green pond frog species, Pelophylax nigromaculatus and P. plancyi. RESULTS: DNA sequence data of one mitochondrial and two nuclear genes from an extensive sampling of the two species were collected, and the genealogies of the three genes were constructed and compared. While the two nuclear genes congruently showed mutual reciprocal monophyly of both species, the mitochondrial phylogeny separated a Korean P. nigromaculatus clade, a paraphyletic central China P. plancyi assemblage, and a large well-supported introgression clade. Within the introgression clade, the mitochondrial haplotypes of the two species were mixed together. This reticulated pattern can be most parsimoniously explained by an ancient mitochondrial introgression event from P. plancyi to P. nigromaculatus that occurred at least 1.36 MYA, followed by multiple recent introgression events from P. nigromaculatus back to P. plancyi within the last 0.63 MY. The re-constitution of previously co-adapted genomes in P. plancyi may be responsible for the recent rampant introgression events. The Korean P. nigromaculatus clade likely represents the only surviving "true" mitochondrial lineage of P. nigromaculatus, and the central China P. plancyi assemblage likely represents the "original" P. plancyi mitochondrial lineage. Refugia in the Korean Peninsula and central China may have played a significant role in preserving these ancient lineages. CONCLUSIONS: The majority of individuals in the two species have either introgressed (P. nigromaculatus) or reclaimed (P. plancyi) mitochondrial genomes while no trace of past hybridization in their nuclear genomes was detected. Asymmetrical reproductive ability of hybrids and continuous backcrossing are likely responsible for the observed mitochondrial introgression. This case is unique in that it includes an ancient "forward" introgression and many recent "backward" introgressions, which re-constitutes the original nuclear and mitochondrial genomes of P. plancyi. This hybrid system provides an excellent opportunity to study cyto-nuclear interaction and co-adaptation.

The female genitalic morphology of "micronetine" spiders (Araneae, Linyphiidae).

Current knowledge of "micronetine" female genitalia is almost exclusively based on transmitted light microscopy data. As such, our understanding of the epigynal anatomy is incomplete and somewhat misleading, to the extent that it hinders comparative studies of linyphiid diversity. We used scanning electronic microscopy (SEM) to study the complex epigynal morphology of "micronetine" spiders. Enzymatic digestion of soft tissues allowed us to examine the internal chitinized structures in detail using SEM. A taxonomic sample of nine species was selected to represent the morphological genitalic diversity of female "micronetines" (including one member of the Erigoninae clade). Results reveal that the epigynum consists of a pair of grooves formed by integument folds (copulatory and fertilization grooves). The protruding epigynal region is divided into a ventral and a dorsal plate by the grooves; both plates can be modified to form an epigynal cavity and/or a scape. Our observations confirm the widespread occurrence of epigynal grooves, rather than ducts, in "micronetines". Epigynal grooves seem to be common in linyphioids and other spider groups.