Search

The genus Bartonella (Family: Bartonellaceae ; Order: Rhizobiales; Class: Alphaproteobacteria) comprises facultative intracellular Gram-negative, haemotropic, slow-growing, vector-borne bacteria. Wild rodents and their fleas harbor a great diversity of species and strains of the genus Bartonella , including several zoonotic ones. This genetic diversity coupled with a fastidious nature of the organism results in a taxonomic challenge that has led to a massive collection of uncharacterized strains. Here, we report the genomic and phenotypic characterization of two strains, members of the genus Bartonella (namely Tel Aviv and OE 1–1), isolated from Rattus rattus rats and Synosternus cleopatrae fleas, respectively. Scanning electron microscopy revealed rod-shaped bacteria with polar pili, lengths ranging from 1.0 to 2.0 µm and widths ranging from 0.3 to 0.6 µm. OE 1–1 and Tel Aviv strains contained one single chromosome of 2.16 and 2.23 Mbp and one plasmid of 29.0 and 41.5 Kbp, with average DNA G+C contents of 38.16 and 38.47 mol%, respectively. These strains presented an average nucleotide identity (ANI) of 89.9 %. Bartonella elizabethae was found to be the closest phylogenetic relative of both strains (ANI=90.9–93.6 %). The major fatty acids identified in both strains were C18:1ω7c, C18 : 0 and C16 : 0. They differ from B. elizabethae in their C17 : 0 and C15 : 0 compositions. Both strains are strictly capnophilic and their biochemical profiles resembled those of species of the genus Bartonella with validly published names, whereas differences in arylamidase activities partially assisted in their speciation. Genomic and phenotypic differences demonstrate that OE 1–1 and Tel Aviv strains represent novel individual species, closely related to B. elizabethae , for which we propose the names Bartonella kosoyi sp. nov. and Bartonella krasnovii sp. nov.

Bartonella effector proteins (named Beps) are substrates of VirB type IV secretion system for translocation into host cells evolved in Bartonella spp. Among these, BepE has been shown to protect cells from fragmentation effects triggered by other Beps and to promote in vivo dissemination of bacteria from the dermal site of inoculation to the bloodstream. Bacterial pathogens secreted effectors to modulate the interplay with host autophagy, either to combat autophagy to escape its bactericidal effect or to exploit autophagy to benefit intracellular replication. Here, we reported a distinct phenotype that selective autophagy in host cells is activated as a countermeasure, to attack BepE via conjugation with K63 polyubiquitin chain on BepE. We found that ectopic expression of Bartonella quintana BepE specifically induced punctate structures that colocalised with an autophagy marker (LC3-II) in host cells, in addition to filopodia and membrane ruffle formation. Two tandemly arranged Bartonella Intracellular Delivery (BID) domains in the BepE C-terminus, where ubiquitination of sister pairs of lysine residues was confirmed, were essential to activate host cell autophagy. Multiple polyubiquitin chain linkages of K27, K29, K33, and K63 were found to be conjugated at sites of K222 and K365 on BepE, of which K63 polyubiquitination on BepE K365 determined the selective autophagy (p62/SQSTM1 positive autophagy) independent of the PI3K pathway. Colocalisation of BepE with LAMP1 confirmed the maturation of BepE-induced autophagosomes in which BepE were targeted for degradation. Moreover, host cells employed selective autophagy to counter-attack BepE to rescue cells from BepE-induced endocytosis deficiency.

BartonellaT4SS effector BepC was reported to mediate internalization of bigBartonellaaggregates into host cells by modulating F-actin polymerization. After that, BepC was indicated to induce host cell fragmentation, an interesting cell phenotype that is characterized by failure of rear-end retraction during cell migration, and subsequent dragging and fragmentation of cells. Here, we found that expression of BepC resulted in significant stress fiber formation and contractile cell morphology, which depended on combination of the N-terminus FIC (filamentationinduced byc-AMP) domain and C-terminus BID (Bartonellaintracellulardelivery) domain of BepC. The FIC domain played a key role in BepC-induced stress fiber formation and cell fragmentation because deletion of FIC signature motif or mutation of two conserved amino acid residues abolished BepC-induced cell fragmentation. Immunoprecipitation confirmed the interaction of BepC with GEF-H1 (a microtubule-associated RhoA guanosine exchange factor), and siRNA-mediated depletion of GEF-H1 prevented BepC-induced stress fiber formation. Interaction with BepC caused the dissociation of GEF-H1 from microtubules and activation of RhoA to induce formation of stress fibers. The ROCK (Rho-associated protein kinase) inhibitor Y27632 completely blocked BepC effects on stress fiber formation and cell contractility. Moreover, stress fiber formation by BepC increased the stability of focal adhesions, which consequently impeded rear-edge detachment. Overall, our study revealed that BepC-induced stress fiber formation was achieved through the GEF-H1/RhoA/ROCK pathway.

Bartonella effector proteins (named Beps) are substrates of VirB type IV secretion system for translocation into host cells evolved in Bartonella spp. Among these, BepE has been shown to protect cells from fragmentation effects triggered by other Beps and to promote in vivo dissemination of bacteria from the dermal site of inoculation to the bloodstream. Bacterial pathogens secreted effectors to modulate the interplay with host autophagy, either to combat autophagy to escape its bactericidal effect or to exploit autophagy to benefit intracellular replication. Here, we reported a distinct phenotype that selective autophagy in host cells is activated as a countermeasure, to attack BepE via conjugation with K63 polyubiquitin chain on BepE. We found that ectopic expression of Bartonella quintana BepE specifically induced punctate structures that colocalised with an autophagy marker (LC3-II) in host cells, in addition to filopodia and membrane ruffle formation. Two tandemly arranged Bartonella Intracellular Delivery (BID) domains in the BepE C-terminus, where ubiquitination of sister pairs of lysine residues was confirmed, were essential to activate host cell autophagy. Multiple polyubiquitin chain linkages of K27, K29, K33, and K63 were found to be conjugated at sites of K222 and K365 on BepE, of which K63 polyubiquitination on BepE K365 determined the selective autophagy (p62/SQSTM1 positive autophagy) independent of the PI3K pathway. Colocalisation of BepE with LAMP1 confirmed the maturation of BepE-induced autophagosomes in which BepE were targeted for degradation. Moreover, host cells employed selective autophagy to counter-attack BepE to rescue cells from BepE-induced endocytosis deficiency.

Sepsis is a life-threatening condition that may develop to multiple organ failure and septic shock. Autophagy is considered to play an important role in the regulation of inflammation. The present study aims to investigate the protective role of mTORC1 inhibitor, rapamycin, on septic death using cecal ligation and puncture (CLP) mice model. Here, results showed that pretreatment with rapamycin reduced the pyroptosis of peritoneal macrophages stimulated by cecal contents and the release of inflammatory factors such as interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α); In septic mice, rapamycin treatment decreased the activation of inflammasome in lung, and alleviated the pathological injuries in lung, liver and spleen tissues during acute stage of sepsis. Treatment of rapamycin rescued animals from septic death significantly. Our results indicated that activation of autophagy is a potential strategy to regulate the excessive inflammation in acute stage of sepsis.

A high-performance capacitor battery AC/λ-MnO2 based on Na-ion insertion-deinsertion into λ-MnO2 in 1 mol dm−3 Na2SO4 solution is successfully demonstrated. A simple acid leaching technique is used to produce λ-MnO2 by treating LiMn2O4 with H2SO4. X-ray power diffraction and inductively coupled plasma analysis provide supporting evidences for the extraction of Li+ from the LiMn2O4, and the particle morphology of λ-MnO2 is studied by scanning and transmission electron microscopy. A high specific capacity of 390.7 mAh g−1 is obtained for the λ-MnO2 electrode at a current density of 13.6 mA g−1 with a nearly 100% efficiency. The capacitor battery AC/λ-MnO2 can operate at a cell voltage as high as 2.2 V, and it exhibits a specific energy of 19.7 Wh Kg−1 at a high power density of 3500 W Kg−1 and a high energy density of 71.7 Wh Kg−1 at a low power density of 403 W Kg−1. It has potentials to be used as an energy storage device for the integration of solar and wind power into the electrical power grid.

In this study, a metabolomics approach based on nuclear magnetic resonance spectroscopy and pertinent multivariate data analyses was used to evaluate the effect of ketamine on metabolic markers in cynomolgus macaques. Principal component analysis and orthogonal projection to latent structure with discriminant analysis showed that ketamine (10 mg/kg) induced metabolic perturbations. Compared with the control group, ketamine-treated macaques had lower serum levels of α-glucose, myoinositol, lactate and succinate and lower urine levels of pyruvate and lactate. In contrast, the levels of leucine in serum and arginine in urine were significantly higher in the ketamine group. Our results also demonstrated that a single injection of ketamine influenced the major energy and amino acid metabolic pathways in cynomolgus macaques. Our study suggests that these influences should be considered in the design of experiments and the interpretation related blood and urine data from ketamine-sedated cynomolgus macaques.

Highlights • We developed a fast and accurate RT-LAMP detection method for Porcine Sapelovirus. • The amplification can be carried out at consistent 63 °C in a water bath. • The sensitivity of this method was 10 copies/μL. • The primers were specific to Porcine Sapelovirus with no cross-reaction with other related viruses. • The RT-LAMP for detection of Porcine Sapelovirus was better than SYBR RT-PCR and/or conventional RT-PCR., The present study describes the development and validation of a one-step, single-tube, and real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) detecting Porcine Sapelovirus. RT-LAMP characterized by one strand displacement reaction with the specific stem-loop structure and Bst DNA polymerase could be finished in 60 min under isothermal condition at 63 °C. RT-LAMP assay showed higher sensitivity with 101 copies/μL than RT-PCR for the detection of Sapelovirus. The specificity of RT-LAMP assay was validated by the absence of any cross-reaction with other closely related virus in Picornaviridae group and other common virus causing porcine diarrhea. 7 positive Sapelovirus infection out of 63 fecal samples were identified using RT-LAMP, while 5 positive samples were determined by a conventional RT-PCR. A cost-effective method for Saplovirus detection with high sensitivity and specificity was developed and evaluated.

Five pathogen-free miniature pigs (minipigs) were infected with the virulent strain SH08 of Streptococcus suis 2 (SS2) by intramuscular injection. The pigs died consecutively within 72 h after the challenge. An additional five non-infected pigs were euthanised and used as controls. Microstructural observations showed that degeneration, bleeding, congestion, cellular necrosis, and an increase in inflammatory cells were present in all organs and tissues except the brain. Ultrastructural observations revealed mitochondrial vacuolation and malformed or missing cristae, indicating that infection of minipigs with strain SH08 of SS2 can lead to extensive lesions in major internal organs and tissues. The findings also demonstrated that the minipig is a useful model for the study of SS2 infection.

A simple acid leaching technique is demonstrated to produce λ-MnO2 by treating LiMn2O4 with H2SO4. λ-MnO2 shows high intercalation capacities as the positive electrode material for rechargeable aqueous batteries. The structure and morphology of the nano-particles are characterized by X-ray power diffraction, inductively coupled plasma, X-ray photoelectron spectroscopy, scanning and transmission electron microscopy. The electrochemical performance of λ-MnO2 is investigated by cyclic voltammetry, constant current charge-discharge tests and electrochemical impedance spectroscopy. λ-MnO2 has ideal battery behaviors for polyvalent cations (Mg2+, Zn2+) to intercalate/deintercalate in the aqueous electrolyte. The λ-MnO2 electrode shows a high initial discharge capacity of 545.6 mAh g−1 at 13.6 mA g−1 in 0.5 mol dm−3 MgCl2 with a columbic efficiency of nearly 100%. In addition, it shows good cycling stability and maintains a capacity of 155.6 mAh g−1 after 300 cycles in 1 mol dm−3 MgCl2 electrolyte.

Magnesium ferrite nanoparticles are prepared by a sol–gel method and characterized by X-ray diffraction, scanning electron microscopy and transmission electron microscopy. The electrochemical behavior as anode materials of lithium-ion rechargeable batteries is investigated by galvanostatic discharge/charge measurements and cyclic voltammetry. The average diameter of the MgFe 2 O 4 particles is around 11 nm. MgFe 2 O 4 shows better performance than the traditional carbon material. The initial discharge specific capacity under the current density of 180 mA g −1 reaches 1404 mAh g −1 , which is higher than that reported in literatures. After 50 cycles, the discharge and charge capacity of the MgFe 2 O 4 under 90 mA g −1 is 493 and 473.6 mAh g −1 , and the irreversible capacity loss is less than 5.2%.

Nanostructured CoxNi1−x(OH)2 (x = 0, 0.053, 0.068, 0.116, 0.192) composites are synthesized via a simple template-free growth process onto nickel foam by using a mixed aqueous solution of nickel nitrate, cobalt nitrate and ammonia. Their structure and surface morphology are characterized by X-ray diffraction, energy dispersive X-ray spectroscopy and scanning electron microscopy. The SEM images show changes in the microstructure of β-Ni(OH)2 by the doping of cobalt. The effects of cobalt content on the electrochemical behaviors of β-Ni(OH)2 are investigated by cyclic voltammetries, galvanostatic charge/discharge and electrochemical impedance spectroscopy. The results show that the Co0.116Ni0.884(OH)2 composite electrode exhibits a drastic improvement in the capacitive characteristics of β-Ni(OH)2 with a specific capacitance increase from 386.4 to 1082.6 F g−1 at a high discharging current density of 6 A g−1. This work suggests that the as-prepared Co0.116Ni0.884(OH)2 composite electrode has a promising future as higher charging/discharging rate materials for pseudo-supercapacitors.

The hallmark of Bartonella infection is long-lasting intraerythrocytic parasitism. However, the process of Bartonella bacteremia is still enigmatic. In the current study, we used Bartonella tribocorum to determine how Bartonella invasion into the bloodstream from dermal inoculation might occur. Bartonella was poorly phagocytized by peritoneal macrophages in vitro. Intracellular Bartonella survived and replicated in macrophages at an early stage of infection. Intracellular Bartonella inhibited spontaneous cell death of macrophages. They also inhibited Salmonella-induced pyroptosis and mildly reduced inflammasome activation through an unidentified mechanism. A rat model confirmed that Bartonella was also inadequately phagocytized in vivo, because numerous free-floating bacilli were observed in lymph collected from thoracic duct drainage as early as 2 hours after inoculation. Lymphatic fluid drainage in the bloodstream significantly reduced the bacterial load in the bloodstream. These findings illustrated a potential route by which Bartonella invade bloodstream from dermal inoculation before they are competent to infect erythrocytes.

Subclinical pathological changes in the kidneys of broiler chickens and suppression of growth caused by the avian nephritis virus (ANV) affect poultry flocks worldwide. A test for detection of virus-specific antibodies in serum would be useful for epidemiological investigations, however the poor propagation in cell cultures has restricted the development of serological tests based on the use of ANV particles as antigens. An enzyme-linked immunosorbent assay (ELISA) was developed for detection of ANV-specific antibodies in chicken serum, using a recombinant protein antigen prepared by segmentation expression of the capsid protein antigen epitope of ANV (HM029238) transfected into Escherichia coli. The expressed fusion protein was detected by Western blotting with ANV-positive serum, and the optimal immunoreactive fusion P1 protein was determined. Using the optimized P1-ELISA, ANV-specific antibodies were detected in commercial chicken flocks aged 10-25 days obtained from the Liaoning Province, China. Out of 960 serum samples, 459 (47.8%) were positive for infection with ANV. These results indicate that the P1-ELISA is helpful for preliminary serological diagnosis of ANV infection, and could be used to for screening in ANV infection and for determining antibodies against ANV.

Porcine kobuvirus was first identified in 2007 in Hungary. The virus has been detected in several Asian countries. In our study, the complete genome of the recently identified porcine kobuvirus strain SH-W-CHN was amplified by RT-PCR and was sequenced. Dendrograms indicated that SH-W-CHN is closely related to other porcine kobuviruses. To identify sites of possible recombination within the genome of the SH-W-CHN strain, the SimPlot program was used to perform recombination analysis. The results showed that no significant recombination event between strain S-1-HUN and Y-1-CHI had occurred. However, certain possible recombination signals were identified, indicating that some early recombination events may have contributed to the genome of SH-W-CHN. This study further confirmed the existence of multiple lineages of porcine kobuvirus and indicated that homologous recombination may be a driving force in its evolution.

Avian astrovirus infections are widespread in many countries, and infections have been linked to enteritis and increased mortality in young poultry. Although pigeons are treated as an important poultry product in some countries, their diseases are often poorly understood and astrovirus infection in pigeons has not been reported. In the present study, faecal samples were collected during an outbreak of gastrointestinal illness in a population of Shanghai pigeons. The samples were examined for astroviruses by reverse transcription-polymerase chain reaction. Eighty-nine per cent (40/45) and 4% (2/45) were found to be positive for avian nephritis virus (ANV) and chicken astrovirus, respectively. One positive sample indicated a co-infection with both ANV and chicken astrovirus. Phylogenetic analysis based on the partial polymerase gene sequence and full-length capsid protein from published avian astrovirus sequences in GenBank revealed that the pigeon viruses detected in this study were evolutionarily closely related to chicken ANV. The present study provided evidence for the presence of astrovirus in pigeons and suggests that cross-infection between pigeons and commercial chickens was likely. Whether the astroviruses in pigeons were responsible for the diarrhoea remains to be determined.

Pigs are increasingly recognized to harbor a wide range of viruses that apparently establish long-term persistence in these animals. They serve as reservoirs for a number of human zoonotic diseases. In this study, a porcine astrovirus (PAstV) strain, designated as PAstV JWH-1, is identified from a diarrheal pig in China, and it is partially characterized genetically. Sequence analysis shows that the PAstV JWH-1 strain contains divergent nucleotide sequences in both the open reading frame (ORF)1b/ORF2 consensus and the 3'-UTR regions (s2m motif), which are usually highly conserved among members of the family Astroviridae. Phylogenetic analysis indicates that the JWH-1 strain clusters closely with newly identified strains PAstV 12-4 and 14-4 and forms a group of mamastroviruses with the proposed novel deer astrovirus. Further recombination analysis shows that two possible interspecies recombination events between porcine and deer astroviruses occurred in the genome of the JWH-1 strain. This study further confirms that multiple lineages are present among PAstVs, and each lineage likely represents an independent origin. Additionally, the possibility of interspecies transmission among PAstVs is also suggested.

In the current study, the complete genome sequence of a member of the family Astroviridae isolated from pigeons was determined through genetic characterization and phylogeny analysis. The isolated genome sequence was proposed to be that of pigeon avian nephritis virus (ANV), whose genome structure and characteristics were similar to previously reported avian astroviruses. The sequenced ssRNA genome comprises 6928 nucleotides, excluding the poly(A) tail, and contains three open reading frames. Phylogenetic analysis using a partial nucleotide sequence of the polymerase gene and the entire amino acid sequence of the full-length capsid protein revealed that pigeon avian nephritis virus is closely related to the previously published ANV, especially to the Japanese G-4260 and Chinese strains. This investigation provides information on the sequence and genetic characteristics of this virus and contributes to a better understanding of pigeon ANV and the possible occurrence of astrovirus transmission between chickens and pigeons.

This study characterized the dynamic distribution of bacteria in the blood of pigs infected with Streptococcus suis serotype 2 using specific primers and a TaqMan probe designed to amplify the highly conserved S. suis serotype 2 glutamate dehydrogenase (GDH) gene sequences. Gene copy numbers were used to determine the concentration of bacteria in the blood of infected pigs over time using established TaqMan real-time quantitative PCR methodologies (RT-qPCR). The results showed that the detection limit of the RT-qPCR was 10 GDH gene copies. The advantages of utilizing this approach are the high levels of specificity, sensitivity and reproducibility. Bacteria were detected in the blood of infected pigs after 24 h post infection and S. suis GDH gene copies in the experimental group were highest (10(4.15)) on day 7 post infection. Data presented in this report demonstrate that the TaqMan RT-qPCR detection method can be used to characterize the dynamic changes occurring during S. suis serotype 2 blood infections in Bama minipigs thereby facilitating research associated with defining pathogenic mechanisms associated with this organism.

SUMMARYHepatitis E virus (HEV) is a zoonotic pathogen of which several species of animals are considered to be reservoirs. Thirty-eight faecal samples, obtained from 22 species of animals including birds in a wildlife first-aid centre in Eastern China, were tested for HEV RNA. Our survey revealed that in total 28·9% (95% confidence interval 14·5–43·4) of the faecal samples from various mammals and birds were HEV RNA positive. Sequence and phylogenetic analyses of the 11 isolates demonstrated that all sequences clustered in genotype 4 with 96–100% identity to each other. In addition, serum samples from seven animal handlers have shown that five (71·4%) were seropositive. The findings imply that cross-species infection of HEV had probably occurred in this zoo-like location, and moreover, birds can be infected naturally with mammalian HEV.

The aim was to study the relationship between the infection rate of Eperythrozoon suis and the status of the immune system. Four pigs with chronic eperythrozoonosis were divided into experimental and control groups. Immunosuppression of the experimental group was accomplished by injecting with high-dose glucocorticoid (dexamethasone) on 3 consecutive days. Microscopic examination of blood smears was performed to observe the change of infection rate after administration. The blood samples of each group were collected on day 7 after administration for half-nested PCR assay. The microscopic results showed that the infection rate of the experimental group rose distinctly by 40 h late and remained at 90% in the days following administration. No changes were observed in the control group. PCR results showed a single expected 250 bp fragment in each group, which validated the positive microscopic results. This study demonstrated the significant effect of dexamethasone (DEX) on Eperythrozoon infection.

Teschoviruses are widely endemic and commonly found in pig fecal samples. In this study, we collected fecal specimens from various pig herds and genotyped them based on the VP1 gene. Of 322 samples, 276 were positive, giving a PTV infectivity rate of 85.7 %. PTV4 was the most common serotype found in Shanghai, followed by PTV8 and PTV10. Interestingly, Some Shanghai strains belonging to a new PTV serotype were also isolated. In phylogenetic analysis, PTV SH8 did not correspond to any known serotype. PTV4 and PTV6 showed similar levels of sequence identity to PTV SH8. These data suggest that PTV SH8 is a new serotype, distinct from the new serotype PTV wild boar/WB2C-TV/2011/HUN, which clusters with PTV SH2, SH10, and SH25.

To explore the bias between the real pressure and the measured values when handheld pressure gauge (HPG) was used to monitor intermittently the pressure in the intubation balloon, so as to provide some measures for the correct use of HPG.In the first part of the study, HPG was used to measure the pressure with the balloon connected with a three-way tube with which to control the inflation and deflation in a laboratory to measure the pressure in the air bag. After gaining the deviation in this in vitro experiment, it was tested and verified in vivo in adult patients undergoing endotracheal intubation.After 132 times of measurements, it was found that measurement with a HPG might result in an "inherent loss" (3.928±0.291) cmH2O (1 cmH2O=0.098 kPa, t=155.273, P=0.000) between inflation value [(30.000±0.000) cmH2O] and measured value [(26.072±0.291) cmH2O]. In addition, after 214 times repeated measurements, the pressure "loss" during disconnection of the gauge was as high as (1.196±0.954) cmH2O (t=18.348, P=0.000) between filled values [(30.000±0.000) cmH2O] and measured values [(28.804±0.954) cmH2O] and it was named as "error loss". At last, the total error was verified by clinical test, and it was (5.270±2.583) cmH2O (t=29.632, P=0.000) between pressure of filled value [(30.000±0.000) cmH2O] and measured value [(24.730±2.583) cmH2O].When the balloon pressure was monitored intermittently with HPG, the real value should be the measured value plus the "error". In addition, subglottic aspiration should be done before the connection of the balloon to the gauge to prevent the secretions on the cuff falls into the deeper airway, and to maintain the cuff pressure at 30 cmH2O.

Background: Hepatitis E virus (HEV) is a major causative agent of acute clinical hepatitis in adults through much of Asia, the Middle East and Africa. Open reading frame 3 (ORF3) encodes around 120 amino acids of phosphorylation protein that associates with the cytoskeleton, while its precise biological function is still unknown. Objectives: In order to understand the function of ORF3 protein (pORF3) in depth, HEV ORF3 interacting proteins were screened in human hepatocytes cDNA library using two-hybrid system techniques and further verification of the interactions were carried out through co-immunoprecipitation (Co-IP). Materials and Methods: The Cyto-Trap two-hybrid system technology, a classical method for analyzing protein interactions, was used to screen the pORF3 interacting proteins from human hepatocytes cDNA library. Results: Through the Cyto-Trap two-hybrid system, eight proteins interacting with pORF3 were winnowed. The Co-IP results confirmed that hepsin which is reported to function as the inhibitor of several tumors reacted with pORF3. Conclusions: Out of eight screened proteins interacting with pORF3, hepsin was confirmed to have specific interactions with pORF3.

In this study, a total of 187 stool specimens were collected from a pig farm in Hunan province of China, from November 2011 to June 2012. 39 (20.9 %) stool specimens were positive for picobirnaviruses using reverse transcription-polymerase chain reaction. Among 39 stool specimens, 84.6 % (33/39) were identified to be genogroup I (prototype 1-CHN-97), 38.5 % (15/39) belonged to genogroup II (prototype 4-GA-91), and 23.1 % (9/39) of which showed the evidence of genogroup I picobirnavirus were also positive for genogroup II picobirnaviruses. Picobirnaviruses exist in pigs which were divided into five groups according to the age and physiological status. Nineteen representative strains of genogroup I picobirnaviruses and eleven strains of genogroup II picobirnaviruses detected in this study were selected to analyze their phylogenetic relationships with other picobirnaviruses reference strains. The phylogenetic tree analysis suggested the prevalence of multiple picobirnaviruses in pigs in China.

Multiwall carbon nanotubes supported Pd–Co electrocatalysts (Pd–Co/MWCNTs) for hydrazine electrooxidation in alkaline electrolytes were prepared. Their morphology and structure were characterized by scanning electron microscopy, transmission electron microscope, and X-ray diffraction. The effect of the mass ratio of Pd to Co on the catalytic performance was examined via cyclic voltammetric and chronoamperometric measurements. The Pd–Co/MWCNTs with a mass ratio of 1:1 for Pd:Co shows higher catalytic performance than both Pd/MWCNTs and Co/MWCNTs and it has good stability for catalyzing the electrooxidation of hydrazine. The gas collection measurements indicated that hydrazine electrooxidation on Pd–Co/MWCNTs proceeded via a near 4-electron pathway.

Bartonella henselae is capable of invading epithelial and endothelial cells by modulating the function of actin-dependent cytoskeleton proteins. Although understanding of the pathogenesis has been increased by the development of an in vitro infection model involving endothelial cells, little is known about the mechanism of interaction between B. henselae and epithelial cells. This study aims to identify the binding candidates of B. henselae in epithelial cells and explores their effect on B. henselae infection. Pull-down assays and mass spectrometry analysis confirmed that some of the binding proteins (keratin 14, keratin 6, and F-actin) are cytoskeleton associated. B. henselae infection significantly induces the expression of the cytokeratin genes. Chemical disruption of the keratin network by using ethylene glycol tetraacetic acid promotes the intracellular persistence of B. henselae in HeLa cells. However, cytochalasin B and phalloidin treatment inhibits B. henselae invasion. Immunofluorescent staining demonstrates that B. henselae infection induces an F-actin-dependent rearrangement of the cytoskeleton. However, we demonstrated via immunofluorescent staining and whole-mount cell electron microscopy that keratin intermediate filaments are depolymerized by B. henselae. The results indicate that B. henselae achieves an intracellular persistence in epithelial cells through the depolymerization of cytokeratin intermediate filaments that are protective against B. henselae invasion.

The hallmark of Bartonella infection is long-lasting intraerythrocytic parasitism. However, the process of Bartonella bacteremia is still enigmatic. In the current study, we used Bartonella tribocorum to determine how Bartonella invasion into the bloodstream from dermal inoculation might occur. Bartonella was poorly phagocytized by peritoneal macrophages in vitro. Intracellular Bartonella survived and replicated in macrophages at an early stage of infection. Intracellular Bartonella inhibited spontaneous cell death of macrophages. They also inhibited Salmonella-induced pyroptosis and mildly reduced inflammasome activation through an unidentified mechanism. A rat model confirmed that Bartonella was also inadequately phagocytized in vivo, because numerous free-floating bacilli were observed in lymph collected from thoracic duct drainage as early as 2 hours after inoculation. Lymphatic fluid drainage in the bloodstream significantly reduced the bacterial load in the bloodstream. These findings illustrated a potential route by which Bartonella invade bloodstream from dermal inoculation before they are competent to infect erythrocytes.

Transmissible gastroenteritis virus strain AYU was isolated in Shanghai. The complete genome has a length of 28,582 bp and contains seven open reading frames. Sequence analysis suggested that Shanghai strain AYU and U.S. strain Purdue P115 are derived from a common ancestor, as they have 99.6% similarity at the nucleotide level.

Bartonella infection (Bartonella henselae in particular) is responsible for a widening spectrum of human diseases. The persistent colonization of erythrocytes is a feature of Bartonella infection. Endothelial and epithelial cells are also widely used to study the pathogenesis of bartonellosis in vitro. Exploring a convenient method for visualizing the bacillus without affecting infectivity would be very interesting. Carboxyfluorescein diacetate succinimidyl ester (CFSE) has been previously used for staining several bacterial species to study their adhesion to host cells. The present study demonstrated the efficiency and safety of using CFSE in staining B. henselae. The staining of bacillus-invaded erythrocytes and epithelial cells in vitro successfully allowed for flow cytometry and confocol microscopy analyses. Parallel tests using untreated bacteria confirmed that CFSE staining did not result in side effects on the infectivity of B. henselae. Labeling Bartonella with CFSE is a valuable method for studying the bacteria-host interaction.

In this study, 176 serum specimens were collected from hospitalized cardiovascular diseases patients in Shanghai, China. 140 samples (79.5%) were positive with the primers located in non-coding region, and 30 samples (17.04%) were identified to be positive with group-specific primers. Of the 30 samples, 9 (5.11%) were classified to group 1, 23 (13.07%) were clustered to group 2, and 3 belonged to group 3. Five samples were confirmed to be dual infection with different groups of TTMV, and no sample was found to be infected with groups 4–6.

The genus Bartonella is comprised of 20 species and subspecies, in which 10 species are responsible for human infections. The identification of the causative agent of cat-scratch disease, bacillary angiomatosis, urban trench fever, and Carrion's disease has raised the emerging medical importance of Bartonella spp. This article provides an overview of bartonellosis with emphasis on its prevalence in China. Key words: Bartonella, cat-scratch disease, wild animal, prevalence.

This study established a SYBR Green I real-time quantitative PCR detection assay (RT-qPCR) for the detection of the porcine transmissible gastroenteritis virus (TGEV) using specific primers designed to amplify the highly conserved porcine TGEV S gene sequence. The threshold cycle (Ct) and the log plasmid copy numbers had a good linear relationship with an efficiency of 1.05 (R2 = 0.999). The advantages of utilizing this approach for the rapid detection of TGEV include excellent sensitivity, reproducibility, and low cost. Ninety-six porcine fecal samples were tested in this study, and 7 samples more than PCR assay were detected by this assay.

With the improvements in diagnostic techniques, Bartonella henselae (B. henselae) infection has recently been recognized to cause a widening spectrum of diseases. Cats are the natural reservoir hosts of B. henselae. The current study aims to investigate the prevalence of B. henselae infection in the cat populations in China. Polymerase chain reaction (PCR) and bacterial cultures confirm that 12.7% of the tested cats were positive for the infection. Old age and outdoor exposure were statistically associated with the infection. Multilocus sequence typing and eBURST analysis of the cat isolates collected in the present study show that 65.4% of the isolates belong to sequence type 1 (ST1). Three new STs (ST16–18) were identified in Midwestern China. These results may aid our understanding of the population structure of B. henselae in China and the relationship between human and cat strains in subsequent studies., Author Summary Bartonella infection (Bartonella henselae in paticular) is responsible for a widening spectrum of human diseases. Cats are thought as the major source of human infections. In most of the developing countries, however, this infection lacks recognition especially in clinicians. Some well known bartonelloses, like cat-scratch disease, bacillary angiomatosis, and infective endocarditis had not been clinically noticed in these countries. In the present study, the prevalence of Bartonella infection in cat populations was investigated in mainland China. As a result, 12.7% of cats were confirmed positive with Bartonella infection. Factors of age, gender, outdoor experience, and geographic location were analyzed in association with infection. The authors found old cats and stray cats are susceptible populations. Some new molecular types are prevalent in Mid-Western China. Inclusion of these types also changed the characteristics of original molecular structure. This research provides the first molecular investigation of Bartonella infection in cats in China, and promotes our understanding of the relationship between human and cat strains in subsequent studies.

Investigations were carried out to identify the causative agent of acute diarrhea, respiratory distress, and polioencephalomyelitis of pigs on a swine farm in Shanghai, China. Samples from the affected animals were tested for viruses and bacteria that are known to cause similar symptoms in swine, and only porcine sapelovirus (PSV; designated as csh strain) was isolated. The presence of PSV was further confirmed by the specific cytopathic effects observed in susceptible cells and by the results of PCR and electron microscopy. Nucleotide sequencing and phylogenetic analysis showed that this isolate is PSV. When inoculated into healthy pigs, PSV.csh caused the same symptoms as observed in the affected herd. Therefore, PSV.csh is the causative agent of this disease. To the best of our knowledge, this is the first report of PSV infecting piglets in China.

In the present study, two isolates (SH-F1 and SH-F2) of Torque teno felis virus (feline TTV) were detected in 16 (12.5%) serum samples collected from cats in China. Their full length genomes were cloned and sequenced. The results showed that they were 2063bp in length and contained three open reading frames (ORFs) (ORF1: nt438-1748, ORF2: nt268-585 and ORF3: nt268-581, 1461-1842). Phylogenetic analysis showed that they were clustered with the strain of Japan (Fc-TTV4, AB076003) and the strain of France (PRA4, EF538878). Sequence analysis indicated that SH-F1 had high (97.5% and 93.3%) identity with the strain of Japan and the strain of France, and SH-F2 shared 94.5% and 92.1% homology with them, respectively. In conclusion, we demonstrated that feline TTV is present in China.

A pig from a commercial pig farm was confirmed to be infected with hemoplasma by microscopic examination in Shanghai, China. However, PCR assay was always negative by using primers that specific of Mycoplasma suis. Therefore, primers that amplified the conserved region of 16S rDNA gene were designed. Phylogenic tree was constructed based on the resulting 16S rDNA sequence, which showed the present hemoplamsa strain is highly similar with Mycoplasma wenyonii (M. wenyonii). This is the first report describing the infection of M. wenyonii in swine blood, which indicated the occurrence of interspecies transmission of hemoplasma is more frequent than our previous thought.

This study aimed to study the immune activity of glutamate dehydrogenase (GDH) of streptococcus suis serotype 2 and detect its activity. METHODS: The GDH gene was cloned and sequenced. Recombinant GDH protein was expressed in a prokaryotic expression system and purified by affinity chromatography. Immunocompetenc of the resulting protein was analyzed by western blotting with anti-GDH monoclonal antibody. RESULTS: The fusion protein was efficiently expressed. Western blotting analysis confirmed that the resulting protein was immunogenic. CONCLUSIONS: The GDH gene of S.suis 2 was successfully cloned and the high expressed, which facilitated the further studies on the development of immunological detection and vaccine design.

By using the PAGE (Poly Acrylamide Gel Electrophoresis), the effect of water-borne Cu2+ on the isozymes in gills of E. sinensis was studied. The experimental groups were treated with different concentration of Cu2+ as 0.01, 0.10, 1.00, and 5.00 mg/l, respectively. The results showed that different isozymes had specific responses. The activity of LDH and MDH decreased gradually along with the increasing of Cu2+ level. Increasing of Cu2+ level followed by decreasing of ALP was observed, and the variation of EST wasn't significant. These isozymes variations reflected the toxic effects of Cu2+ sensitively.