Adaptive response of Pseudomonas aeruginosa under serial ciprofloxacin exposure.

Understanding the evolution of antibiotic resistance is important for combating drug-resistant bacteria. In this work, we investigated the adaptive response of Pseudomonas aeruginosa to ciprofloxacin. Ciprofloxacin-susceptible P. aeruginosa ATCC 9027, CIP-E1 (P. aeruginosa ATCC 9027 exposed to ciprofloxacin for 14 days) and CIP-E2 (CIP-E1 cultured in antibiotic-free broth for 10 days) were compared. Phenotypic responses including cell morphology, antibiotic susceptibility, and production of pyoverdine, pyocyanin and rhamnolipid were assessed. Proteomic responses were evaluated using comparative iTRAQ labelling LC-MS/MS to identify differentially expressed proteins (DEPs). Expression of associated genes coding for notable DEPs and their related regulatory genes were checked using quantitative reverse transcriptase PCR. CIP-E1 displayed a heterogeneous morphology, featuring both filamentous cells and cells with reduced length and width. By contrast, although filaments were not present, CIP-E2 still exhibited size reduction. Considering the MIC values, ciprofloxacin-exposed strains developed resistance to fluoroquinolone antibiotics but maintained susceptibility to other antibiotic classes, except for carbapenems. Pyoverdine and pyocyanin production showed insignificant decreases, whereas there was a significant decrease in rhamnolipid production. A total of 1039 proteins were identified, of which approximately 25 % were DEPs. In general, there were more downregulated proteins than upregulated proteins. Noted changes included decreased OprD and PilP, and increased MexEF-OprN, MvaT and Vfr, as well as proteins of ribosome machinery and metabolism clusters. Gene expression analysis confirmed the proteomic data and indicated the downregulation of rpoB and rpoS. In summary, the response to CIP involved approximately a quarter of the proteome, primarily associated with ribosome machinery and metabolic processes. Potential targets for bacterial interference encompassed outer membrane proteins and global regulators, such as MvaT.

Proteomic analysis of ceftazidime and meropenem-exposed Pseudomonas aeruginosa ATCC 9027.

BACKGROUND: Pseudomonas aeruginosa is well known for its intrinsic ability to resist a wide range of antibiotics, thus complicates treatment. Thus, understanding the response of the pathogen to antibiotics is important for developing new therapies. In this study, proteomic response of P. aeruginosa to the commonly used anti-pseudomonas antibiotics, ceftazidime (Caz) and meropenem (Mem) was investigated. METHODS: P. aeruginosa ATCC 9027, an antibiotic-susceptible strain, was exposed to sub-MIC values of antibiotics either Caz or Mem for 14 days to obtain E1 strains and then cultured in antibiotic-free environments for 10 days to obtain E2 strains. Proteomes of the initial and E1, E2 strains were identified and comparatively analyzed using isobaric tags for relative and absolute quantitation (iTRAQ) in cooperation with nano LC-MS/MS. Noted up and down-regulated proteins were confirmed with quantitative reverse transcriptase PCR (qRT-PCR). RESULTS: Overall, 1039 and 1041 proteins were identified in Caz and Mem-exposed strains, respectively. Upon antibiotic exposure, there were 7-10% up-regulated (Caz: 71, Mem: 85) and down-regulated (Caz: 106, Mem: 69) proteins (1.5-fold change cut-off). For both Caz and Mem, the DEPs were primarily the ones involved in metabolic process, membrane, virulence, protein synthesis, and antibiotic resistance in which proteins involved in antibiotics resistance tended to be up-regulated while proteins involved in protein synthesis and metabolic process were down-regulated. Noted proteins included beta-lactamase AmpC which was up-regulated and OprD which was down-regulated in both the antibiotic-exposed strains. Besides, biofilm formation related proteins TssC1 and Hcp1 in Caz- exposed strains and the membrane/ periplasmic proteins Azu and PagL in Mem-exposed strains were found significantly down-regulated. qRT-PCR results confirmed the expression change of AmpC, Hcp1 and OprD proteins. CONCLUSION: Exposure of Pseudomonas aeruginosa to sub-MIC values of Caz and Mem resulted in around 10% change in its proteome. Not only proteins with confirmed roles in antibiotic resistance mechanisms changed their expression but also virulence- associated proteins. Both Caz and Mem response involved up-regulation of AmpC and down-regulation of OprD. While TssC1 and Hcp1 were responsible for Caz response, Azu and PagL were more likely involved in Mem response.

Radiological risk assessment of outdoor222Rn and220Rn around rare earth element and uranium mines from northern Vietnam.

The outdoor222Rn and220Rn concentrations at 320 sampling points at 1 m above the ground in different sites surrounding rare earth element (REE) and uranium mines from northern Vietnam were measured using the RAD7. Results showed that222Rn concentrations were always higher than220Rn concentrations with large variation ranges from 25.7 to 573 Bq m-3and from 18.5 to 385 Bq m-3, respectively. The high correlation between220Rn and228Ra concentrations in surface soil of the studied sites were observed. The highest220Rn and222Rn concentrations are found at the sampling points of the REE NX-Lai Chau site. The220Rn and222Rn activities surrounding the REE mines were found to be higher than those surrounding the uranium mines. The average annual committed effective doses originated from the inhalation of220Rn and222Rn outdoor concentrations is about five times higher than the worldwide average value.

Molecular characterisation of the current high prevalence of the new CPV-2c variants in the Southern Vietnamese dogs signifies a widespread in the worldwide dog population.

BACKGROUND: Canine parvovirus type 2 (CPV-2) is known as the primary etiological agent cause of acute gastroenteritis, myocarditis and death of canids worldwide. In Vietnam, although CPV-2 infection and its outbreaks are the most important risk factors of the canine's health concern, lack of available information about the molecular epidemiology of the CPV-2. OBJECTIVES: In this study, the complete coding sequences of 10 CPV-2 strains collected from dogs vaccinated with CPV-2 vaccination were analysed to better understand the genomic characteristics of the current circulating CPV-2 in Vietnam. METHODS: Ten CPV-specific PCR-positive rectal swab samples were collected from dogs with acute symptoms of haemorrhagic diarrhoea and vomiting in Vietnam in 2019. The complete coding sequences of these CPV strains were analysed to determine their phylogeny and genetic relationship with other available CPV strains globally. RESULTS: Analysis of the VP2 gene sequences demonstrated that the studied strains belonged to the new CPV-2c variants with the unique mutations at amino acids 5Ala-Gly and 447Iso-Met . Phylogenetic tree analysis indicated that the studied strains share a common evolutionary origin with the current CPV-2c strains circulating in dogs in Asia countries, including China, Thailand, Taiwan and Mongolia, in recent years. Low sequence identity between the studied strains and commercial vaccine strains was observed. CONCLUSIONS: This study provides deep insights into the molecular characteristics, genetic diversity, and evolution of circulating CPV-2 strains in Vietnam. We recommend more studies to estimate the effectiveness of the CPV vaccine and the need to continue developing other effective vaccination essential to better control the widespread of these new CPV-2 variants.

Transition of dominant canine parvovirus genotype from 2b to 2c in Vietnamese dogs.

Canine parvovirus (CPV) is one of the most important pathogens causing enteritis in dogs. Although there have been a few reports of CPV in Vietnam, recent information on CPV infection in domestic dogs in Vietnam is limited. Faecal samples collected from 30 diarrheic and 50 healthy dogs were examined by PCR for detection of CPV DNA. The prevalence of CPV in diarrheic dogs (43.3%, 13/30) was significantly higher than in healthy dogs (4.0%, 2/50), indicating that CPV was a major cause of diarrhoea in domestic dogs. Genotyping of 15 CPV strains showed that both CPV­2a and CPV­2c were circulating and that CPV­2c was a dominant CPV variant in Vietnam. Virus isolation was performed from faecal samples using A72/cSLAM cells, and nine CPV strains were successfully isolated. The dominant genotype spreading among Vietnamese dogs has changed from CPV­2b to CPV­2c.

Evaluation of the Safety Profile of the ASFV Vaccine Candidate ASFV-G-ΔI177L.

African swine fever (ASF) is the cause of a recent pandemic that is posing a threat to much of the world swine production. The etiological agent, ASF virus (ASFV), infects domestic and wild swine, producing a variety of clinical presentations depending on the virus strain and the genetic background of the pigs infected. No commercial vaccine is currently available, although recombinant live attenuated vaccine candidates have been shown to be efficacious. In addition to determining efficacy, it is paramount to evaluate the safety profile of a live attenuated vaccine. The presence of residual virulence and the possibility of reversion to virulence are two of the concerns that must be evaluated in the development of live attenuated vaccines. Here we evaluate the safety profile of an efficacious live attenuated vaccine candidate, ASFV-G-ΔI177L. Results from safety studies showed that ASFV-G-ΔI177L remains genetically stable and phenotypically attenuated during a five-passage reversion to virulence study in domestic swine. In addition, large-scale experiments to detect virus shedding and transmission confirmed that even under varying conditions, ASFV-G-ΔI177L is a safe live attenuated vaccine.

African swine fever virus vaccine candidate ASFV-G-ΔI177L efficiently protects European and native pig breeds against circulating Vietnamese field strain.

The recent spread of African swine fever (ASF) in the People's Republic of China and neighbouring countries in Asia has had significant economic consequences with an estimated direct cost of $55-$130 billion. This pandemic, originally detected in Republic of Georgia in 2007, has devastated the swine industry in large geographical areas of Southeast Asia with 14 countries reporting ASF outbreaks since the first documented case was confirmed in the city of Shenyang, Liaoning Province, China, on 3 August 2018. In the absence of any available vaccines, the control of ASF relies on the detection and culling of infected animals. The United States Department of Agriculture recently developed a recombinant experimental vaccine candidate, ASFV-G-ΔI177L, by deleting the I177L gene from the genome of the highly virulent pandemic ASFV strain Georgia, which efficaciouly protects pigs from the parental virus. Here, the initial studies were extended demonstrating that ASFV-G-ΔI177L is able to protect pigs against the virulent ASFV isolate currently circulating and producing disease in Vietnam with similar efficacy as reported against the Georgia strain. Comparative studies performed using a large number of pigs of European and Vietnamese origin demonstrated that a minimum protective dose of 102 HAD50 of ASFV-G-ΔI177L equally protects animals of both breeds. In concurrence with those results, the onset of immunity in these animal breed showed appearance of protection in approximately one-third of the animals by the second week post vaccination, with full protection achieved by the fourth week post vaccination. Therefore, results presented here demonstrated that ASFV-G-ΔI177L is able to induce protection against virulent Vietnameese ASFV field strains and is effective in protecting local breeds of pigs as efficiently as previously shown for European cross-bred pigs. To our knowledge, this is the first report showing the efficacy of a Georgia 2007 based vaccine candidate in Asian breed of pigs or challenged with an Asian ASFV strain.

Prevalence and Virulence of Commensal Pseudomonas Aeruginosa Isolates from Healthy Individuals in Southern Vietnam (2018-2020).

Understanding the colonization of Pseudomonas aeruginosa (P. aeruginosa) in healthy humans is useful for future prevention and treatment of P. aeruginosa infection. This study aimed to investigate the prevalence and risk factors of of P. aeruginosa colonization in healthy humans. At the same time, the virulence of the isolated P. aeruginosa was also studied. In the study, 609 Vietnamese volunteers (310 females and 299 males, age range of 2 to 73 years), who had no acute infection or disease symptoms participated at the time of sample collection. Samples were taken from the throat, nostrils, and outer ears. P. aeruginosa was found in 19 participants (3.12%, 95% CI: 0.017−0.045), mainly from the throat (11/19, 57.89%). Participants with a history of sinusitis were 11.57 times more likely to be colonized with P. aeruginosa than participants without a history of sinusitis (OR: 11.57, 95% CI: 4.08−32.76, p-value < 0.0001, Fisher's Exact test). Age and sex were not significantly associated with P. aeruginosa colonization. Among 16 P. aeruginosa isolates used in virulence tests, 100% (16/16) were positive for the synthesis of biofilm, pyocyanin, and siderophores; 93.75% (15/16) isolates were positive for the synthesis of gelatinase and protease; and 50% (8/16) isolates were positive for lipase. There were no differences in the pattern and range of virulence factors of P. aeruginosa isolates taken from participants with and without sinusitis history. P. aeruginosa colonized 3.12% of participants, and its presence was associated with sinusitis history.

Tomographic density imaging using modified DF-DBIM approach.

Ultrasonic computed tomography based on back scattering theory is the most powerful and accurate tool in ultrasound based imaging approaches because it is capable of providing quantitative information about the imaged target and detects very small targets. The duple-frequency distorted Born iterative method (DF-DBIM), which uses density information along with sound contrast for imaging, is a promising approach for imaging targets at the level of biological tissues. With two frequencies f1 (low) and f2 (high) through N f 1 and N f 2 iterations respectively, this method is used to estimate target density along with sound contrast. The implications of duple-frequency fusion for the image reconstruction quality of density information along with sound contrast based ultrasound tomography have been analyzed in this paper. In this paper, we concentrate on the selection of parameters that is supposed to be the best to improve the reconstruction quality of ultrasound tomography. When there are restraints imposed on simulated scenarios to have control of the computational cost, the iteration number N f 1 is determined resulting in giving the best performance. The DF-DBIM is only effective if there are a moderate number of iterations, transmitters and receivers. In case that the number of transducers is either too large or too small, a result of reconstruction which is better than that of the single frequency approach is not produced by the implementation of DF-DBIM. A fixed sum N iter of N f 1 and N f 2 was given, the investigation of simulation results shows that the best value of N f 1 is N iter 2 - 1 . The error, when applying this way of choosing the parameters, will be normalized with the reduction of 56.11%, compared to use single frequency as used in the conventional DBIM method. The target density along with sound contrast is used to image targets in this paper. It is a fact that low-frequency offers fine convergence, and high-frequency offers fine spatial resolution. Wherefore, this technique can effectively expand DBIM's applicability to the problem of biological tissue reconstruction. Thanks to the usage of empirical data, this work will be further developed prior to its application in reality.

Two-dimensional complex shear modulus imaging of soft tissues by integration of Algebraic Helmoltz Inversion and LMS filter into dealing with noisy data: a simulation study.

Elasticity and viscosity of soft tissues can be obtained from the complex shear modulus imaging (CSMI). CSMI is often used not only to investigate the structure of tissues but also to detect tumors in tissues. One of the most popular ways to categorize the methods used in CSMI is into quasi-static and dynamic methods. In the dynamic method, a force excitation is used to create the shear wave propagation, and the particle velocities are measured to extract their amplitude and phase at spatial locations. These parameters are then employed to directly or indirectly estimate the Complex Shear Modulus (CSM) represented by elasticity and viscosity. Algebraic Helmholtz Inversion (AHI) algorithm provides the direct estimation of CSM using the Finite Difference Time Domain (FDTD) technique. The limitation of this method, however, is that the noise generated from measuring the particle velocity strongly degrades the accuracy of the estimation. To overcome this problem, we proposed in this paper an adaptive AHI (AAHI) algorithm that offers a good performance in CSMI with a mean error of 2.06%.

Weakly Trapped, Charged, and Free Excitons in Single-Layer MoS2 in the Presence of Defects, Strain, and Charged Impurities.

Few- and single-layer MoS2 host substantial densities of defects. They are thought to influence the doping level, the crystal structure, and the binding of electron-hole pairs. We disentangle the concomitant spectroscopic expression of all three effects and identify to what extent they are intrinsic to the material or extrinsic to it, i.e., related to its local environment. We do so by using different sources of MoS2-a natural one and one prepared at high pressure and high temperature-and different substrates bringing varying amounts of charged impurities and by separating the contributions of internal strain and doping in Raman spectra. Photoluminescence unveils various optically active excitonic complexes. We discover a defect-bound state having a low binding energy of 20 meV that does not appear sensitive to strain and doping, unlike charged excitons. Conversely, the defect does not significantly dope or strain MoS2. Scanning tunneling microscopy and density functional theory simulations point to substitutional atoms, presumably individual nitrogen atoms at the sulfur site. Our work shows the way to a systematic understanding of the effect of external and internal fields on the optical properties of two-dimensional materials.

Slaughterhouse pigs are a major reservoir of Streptococcus suis serotype 2 capable of causing human infection in southern Vietnam.

Streptococcus suis is a pathogen of major economic significance to the swine industry and is increasingly recognized as an emerging zoonotic agent in Asia. In Vietnam, S. suis is the leading cause of bacterial meningitis in adult humans. Zoonotic transmission is most frequently associated with serotype 2 strains and occupational exposure to pigs or consumption of infected pork. To gain insight into the role of pigs for human consumption as a reservoir for zoonotic infection in southern Vietnam, we determined the prevalence and diversity of S. suis carriage in healthy slaughterhouse pigs. Nasopharyngeal tonsils were sampled from pigs at slaughterhouses serving six provinces in southern Vietnam and Ho Chi Minh City area from September 2006 to November 2007. Samples were screened by bacterial culture. Isolates of S. suis were serotyped and characterized by multi locus sequence typing (MLST) and pulse field gel electrophoresis (PFGE). Antibiotic susceptibility profiles and associated genetic resistance determinants, and the presence of putative virulence factors were determined. 41% (222/542) of pigs carried S. suis of one or multiple serotypes. 8% (45/542) carried S. suis serotype 2 which was the most common serotype found (45/317 strains, 14%). 80% of serotype 2 strains belonged to the MLST clonal complex 1,which was previously associated with meningitis cases in Vietnam and outbreaks of severe disease in China in 1998 and 2005. These strains clustered with representative strains isolated from patients with meningitis in PFGE analysis, and showed similar antimicrobial resistance and virulence factor profiles. Slaughterhouse pigs are a major reservoir of S. suis serotype 2 capable of causing human infection in southern Vietnam. Strict hygiene at processing facilities, and health education programs addressing food safety and proper handling of pork should be encouraged.

Risk factors of Streptococcus suis infection in Vietnam. A case-control study.

BACKGROUND: Streptococcus suis infection, an emerging zoonosis, is an increasing public health problem across South East Asia and the most common cause of acute bacterial meningitis in adults in Vietnam. Little is known of the risk factors underlying the disease. METHODS AND FINDINGS: A case-control study with appropriate hospital and matched community controls for each patient was conducted between May 2006 and June 2009. Potential risk factors were assessed using a standardized questionnaire and investigation of throat and rectal S. suis carriage in cases, controls and their pigs, using real-time PCR and culture of swab samples. We recruited 101 cases of S. suis meningitis, 303 hospital controls and 300 community controls. By multivariate analysis, risk factors identified for S. suis infection as compared to either control group included eating "high risk" dishes, including such dishes as undercooked pig blood and pig intestine (OR(1) = 2.22; 95%CI = [1.15-4.28] and OR(2) = 4.44; 95%CI = [2.15-9.15]), occupations related to pigs (OR(1) = 3.84; 95%CI = [1.32-11.11] and OR(2) = 5.52; 95%CI = [1.49-20.39]), and exposures to pigs or pork in the presence of skin injuries (OR(1) = 7.48; 95%CI = [1.97-28.44] and OR(2) = 15.96; 95%CI = [2.97-85.72]). S. suis specific DNA was detected in rectal and throat swabs of 6 patients and was cultured from 2 rectal samples, but was not detected in such samples of 1522 healthy individuals or patients without S. suis infection. CONCLUSIONS: This case control study, the largest prospective epidemiological assessment of this disease, has identified the most important risk factors associated with S. suis bacterial meningitis to be eating 'high risk' dishes popular in parts of Asia, occupational exposure to pigs and pig products, and preparation of pork in the presence of skin lesions. These risk factors can be addressed in public health campaigns aimed at preventing S. suis infection.

Prevalence of antibodies against avian influenza A (H5N1) virus among Cullers and poultry workers in Ho Chi Minh City, 2005.

BACKGROUND: Between 2003 and 2005, highly pathogenic avian influenza A (H5N1) viruses caused large scale outbreaks in poultry in the Ho Chi Minh City area in Vietnam. We studied the prevalence of antibodies against H5N1 in poultry workers and cullers who were active in the program in Ho Chi Minh City in 2004 and 2005. METHODOLOGY/PRINCIPAL FINDINGS: Single sera from 500 poultry workers and poultry cullers exposed to infected birds were tested for antibodies to avian influenza H5N1, using microneutralization assays and hemagglutination inhibition assay with horse blood. All sera tested negative using microneutralization tests. Three samples showed a 1ratio80 titer in the hemagglutination inhibition assay. CONCLUSIONS/SIGNIFICANCE: This study provides additional support for the low transmissibility of clade 1 H5N1 to humans, but limited transmission to highly exposed persons cannot be excluded given the presence of low antibody titers in some individuals.

Microstructure and piezoelectric properties of (Na0.5K0.5)NbO3 lead-free piezoelectric ceramics with V2O5 addition.

Various amounts of Nb(2)O(5) in the (Na(0.5)K(0.5)) NbO(3) (NKN) ceramic were replaced by V(2)O(5) to decrease its sintering temperature to below 950 degrees C. A small V(2)O(5) content resulted in a dense microstructure with an increased grain size for the specimen sintered at 900 degrees C due to the presence of a liquid phase. When V(2)O(5) was added to the NKN ceramics, their orthorhombic-to-tetragonal transition temperature increased from 178 degrees C to around 200 degrees C. However, their Curie temperature decreased from 402 degrees C to around 330 degrees C. The k(p), epsilon(3) (T)/epsilon(0), and Q(m) values increased with V(2)O(5) addition, probably due to the increased density and poling state, which was identified by the phase angle. The specimen with x = 0.05, sintered at 900 degrees C for 8 h, exhibited the following piezoelectric properties: k(p) = 0.32, epsilon(3) (T)/epsilon(0) = 245, d(33) = 120 (pC/N), and Q(m) = 232.

The mortality status of the neonates born at other maternities compared to that at Tu Du Hospital in 2003 year

Study on 229 newborn babies transferred from lower-level hospital into Neonatology Department of Tu Du Hospital. The results showed that: The main causes were respiratory distress, neonatal infection and asphyxia in premature infants, asphyxia and respiratory distress in the term infants. The more premature the newborn infants were, the earlier they were hospitalized. 157/229 infants were transferred into the hospital by rudimentary vehicles, the others were transferred by ambulance in which only 9.6% had oxygen supply. 163/229 patients (71.18%) were admitted in very serious status that required active intensive care and had high mortality rate