Slightly acidic electrolyzed water(SAEW)has proven to be an efficient and novel sanitizer in food and agriculture field.This study assessed the efficacy of SAEW(30 mg/L)at 40℃on the inactivation of foodbome pathogens...Slightly acidic electrolyzed water(SAEW)has proven to be an efficient and novel sanitizer in food and agriculture field.This study assessed the efficacy of SAEW(30 mg/L)at 40℃on the inactivation of foodbome pathogens and detachment of multi-resistant Staphylococcus aureus(MRSA)biofilm.Furthermore.the underlying mechanism of MRS A biofilm under heated SAEW at 40℃treatment on metabolic profiles was investigated.The results showed that the heated SAEW at 40℃significantly effectively against foodbome pathogens of 1.96-7.56(lg(CFU/g))reduction in pork,chicken,spinach,and lettuce.The heated SAEW at 40℃treatment significantly reduced MRS A biofilm cells by 2.41(lg(CFU/cm^(2))).The synergistic effect of SAEW treatment showed intense anti-biofilm activity in decreasing cell density and impairing biofilm cell membranes.Global metabolic response of MRSA biofilms,treated by SAEW at 40℃,revealed the alterations of intracellular metabolites,including amino acids,organic acid,fatty acid,and lipid.Moreover,signaling pathways involved in amino acid metabolism,energy metabolism,nucleotide synthesis,carbohydrate metabolites,and lipid biosynthesis were functionally disrupted by the SAEW at 40℃treatment.As per our knowledge,this is the first research to uncover the potential mechanism of heated SAEW treatment against MRSA biofilm on food contact surface.展开更多
Staphylococcus aureus is a serious foodborne pathogen threatening food safety and public health.Especially the emergence of methicillin-resistant Staphylococcus aureus(MRSA)increased the difficulty of S.aureus treatme...Staphylococcus aureus is a serious foodborne pathogen threatening food safety and public health.Especially the emergence of methicillin-resistant Staphylococcus aureus(MRSA)increased the difficulty of S.aureus treatment.Staphyloxanthin is a crucial virulence factor of S.aureus.Blocking staphyloxanthin production could help the host immune system counteract the invading S.aureus cells.In this study,we first screened for staphyloxanthin inhibitors using a virtual screening method.The outcome of the virtual screening method resulted in the identification of eugenol(300μg/mL),which significantly inhibits the staphyloxanthin production in S.aureus ATCC 29213,S.aureus Newman,MRSA ATCC 43300 and MRSA ATCC BAA1717by 84.2%,63.5%,68.1%,and 79.5%,respectively.The outcome of the growth curve assay,field-emission scanning electron,and confocal laser scanning microscopy analyses confirmed that eugenol at the test concentration did not affect the morphology and growth of S.aureus.Moreover,the survival rate of S.aureus ATCC 29213 and MRSA ATCC 43300 under H_(2)O_(2) pressure decreased to 51.9%and 45.5%in the presence of eugenol,respectively.The quantitative RT-PCR and molecular simulation studies revealed that eugenol targets staphyloxanthin biosynthesis by downregulating the transcription of the crtM gene and inhibiting the activity of the CrtM enzyme.Taken together,we first determined that eugenol was a prominent compound for staphyloxanthin inhibitor to combat S.aureus especially MRSA infections.展开更多
Introduction: Bacterial skin and soft tissue infections (SSTIs) are a cause of frequent inpatient and outpatient care visits whose causative agents are associated with a high antimicrobial resistance burden. For insig...Introduction: Bacterial skin and soft tissue infections (SSTIs) are a cause of frequent inpatient and outpatient care visits whose causative agents are associated with a high antimicrobial resistance burden. For insights on antimicrobial susceptibilities in a rural setting, we examined specimens from suspected SSTIs from two public health facilities in Kenya. We additionally assessed antibiotic use, appropriateness of empiric therapy and risk factors for SSTI. Methodology: Between 2021 and 2023, 265 patients at Kisii and Nyamira County Referral hospitals were enrolled. Wound swabs/aspirates were collected and processed following standard microbiological procedures. Identification and antimicrobial susceptibility were performed using the VITEK 2 Compact platform. Demographic, clinical, and microbiological data were analyzed with R Statistical software. Results: S. aureus was isolated in 16.2% (43/265) of patients with a methicillin resistance (MRSA) proportion of 14% (6/43). While 13/15 drugs elicited susceptibilities ranging from 84% - 100%, penicillin (16%) and trimethoprim-sulfamethoxazole [TMP-SXT] (23%) yielded the lowest susceptibilities. Escherichia coli (n = 33), Klebsiella pneumoniae (n = 8), Pseudomonas aeruginosa (n = 8), and Citrobacter species (n = 4) were the most commonly isolated gram-negative species. Gram-negative strains showed high susceptibilities to most of the tested drugs (71% - 100%) with the exception of ampicillin (18%), TMP-SXT (33%), and first and second generation cephalosporins. Conclusions: The low MRSA prevalence and generally high antibiotic susceptibilities for S. aureus and gram-negative bacteria present opportunities for antibiotic stewardship in the study setting. Diminished susceptibilities against penicillin/ampicillin and TMP-SXT accord with prevailing local data and add a layer of evidence for their cautious empiric use.展开更多
Objective:To determine the antibiotic resistance profile(ARP)of Staphylococcus(S.)aureus isolates and molecular typing of the methicillin-resistant S.aureus(MRSA)isolates from Tuanku Mizan Armed Forces Hospital(TMAFH)...Objective:To determine the antibiotic resistance profile(ARP)of Staphylococcus(S.)aureus isolates and molecular typing of the methicillin-resistant S.aureus(MRSA)isolates from Tuanku Mizan Armed Forces Hospital(TMAFH),Kuala Lumpur.Methods:The ARP and presence of the pvl gene were determined for 209 S.aureus isolates from clinical specimens.Of these,123 were methicillin-susceptible S.aureus(MSSA)isolates and 86 were MRSA isolates.All MRSA isolates were characterized using SCCmec typing and spa typing.Descriptive analysis was performed to compare the demographic data with the phenotypic and genotypic variables of the S.aureus isolates.Results:No vancomycin-intermediate and-resistant S.aureus(VISA and VRSA,respectively)were detected among the study isolates.The MSSA isolates showed low resistance rates to all tested antibiotics,were commonly invasive(28/42,66.7%),and mostly harboured pvl(35/42,83.3%).Meanwhile,MRSA isolates showed high resistance to penicillin(86/86,100%),ampicillin(86/86,100%),sulbactam/ampicillin(86/86,100%),cefuroxime(81/86,94.19%),cefoperazone(76/86,88.37%),azithromycin(56/86,65.12%),and erythromycin(54/86,62.79%).The majority of MRSA isolates were of SCCmec type IVh(65/86,75.58%),spa type t032(55/85,63.95%),and grouped into spaCC-t022(66/85,77.65%).The t032 type was found to be associated with resistance traits to azithromycin and erythromycin(P<0.05).We also found several spa types that are typically associated with hospital-,community-,and livestock-associated MRSA co-existing in our MRSA population.Conclusions:This study reflected the consistent absence of VISA and VRSA and corroborated the clonal shifting of MRSA isolates in the Malaysian MRSA isolates.展开更多
Host-directed therapy(HDT)is an emerging novel approach for treating multidrug-resistant Staphylococcus aureus(S.aureus)infection.Functioning as the indispensable specific cellular receptor for a-toxin(Hla),a-disinteg...Host-directed therapy(HDT)is an emerging novel approach for treating multidrug-resistant Staphylococcus aureus(S.aureus)infection.Functioning as the indispensable specific cellular receptor for a-toxin(Hla),a-disintegrin and metalloproteinase 10(ADAM10)is exploited to accelerate S.aureus infection through diverse mechanisms.The extraordinary contribution of ADAM10 to S.aureus pathogenesis renders it an attractive HDT target for combating S.aureus infection.Our study is the first to demonstrate the indispensable role of ADAM10 in S.aureus-induced necroptosis,and it enhances our knowledge of the role of ADAM10 in S.aureus infection.Using a fluorogenic substrate assay,we further identified kaempferol as a potent ADAM10 inhibitor that effectively protected mice from S.aureus infection by suppressing Hla-mediated barrier disruption and necroptosis.Collectively,our work presents a novel hostdirected therapeutic strategy for using the promising candidate kaempferol to treat S.aureus infection and other diseases relevant to the disordered upregulation of ADAM10.展开更多
Background:Staphylococcus aureus can cause serious infections by secreting many superantigen exotoxins in“carrier”or“pathogenic”states.HLA DQ and HLA DR humanized mice have been used as a small animal model to stu...Background:Staphylococcus aureus can cause serious infections by secreting many superantigen exotoxins in“carrier”or“pathogenic”states.HLA DQ and HLA DR humanized mice have been used as a small animal model to study the role of two molecules during S.aureus infection.However,the contribution of HLA DP to S.aureus infection is unknown yet.Methods:In this study,we have produced HLA DP401 and HLA DRA0101 humanized mice by microinjection of C57BL/6J zygotes.Neo-floxed IAβ+/-mice were crossbred with Ella-Cre and further crossbred with HLA DP401 or HLA-DRA0101 humanized mice.After several rounds of traditional crossbreeding,we finally obtained HLA DP401-IAβ-/-and HLA DRA-IAβ-/-humanized mice,in which human DP401 or DRA0101 molecule was introduced into IAβ-/-mice deficient in endogenous murine MHC classⅡmolecules.A transnasal infection murine model of S.aureus pneumonia was induced in the humanized mice by administering 2×108CFU of S.aureus Newman dropwise into the nasal cavity.The immune responses and histopathology changes were further assessed in lungs in these infected mice.Results:We evaluated the local and systemic effects of S.aureus delivered intranasally in HLA DP401-IAβ-/-and HLA DRA-IAβ-/-transgenic mice.S.aureus Newman infection significantly increased the m RNA level of IL 12p40 in lungs in humanized mice.An increase in IFN-γand IL-6 protein was observed in HLA DRA-IAβ-/-mice.We observed a declining trend in the percentage of F4/80+macrophages in lungs in HLA DP401-IAβ-/-mice and a decreasing ratio of CD4+to CD8+T cells in lungs in IAβ-/-mice and HLA DP401-IAβ-/-mice.A decreasing ratio of Vβ3+to Vβ8+T cells was also found in the lymph node of IAβ-/-mice and HLA DP401-IAβ-/-mice.S.aureus Newman infection resulted in a weaker pathological injury in lungs in IAβ-/-genetic background mice.Conclusion:These humanized mice will be an invaluable mouse model to resolve the pathological mechanism of S.aureus pneumonia and study what role DP molecule plays in S.aureus infection.展开更多
Objective:To describe clinical and epidemiological characteristics,antimicrobial susceptibility and mortality-associated factors of healthcare-associated infections(HCAIs)caused by Staphylococcus(S.)aureus in children...Objective:To describe clinical and epidemiological characteristics,antimicrobial susceptibility and mortality-associated factors of healthcare-associated infections(HCAIs)caused by Staphylococcus(S.)aureus in children.Methods:We conducted a retrospective,single-centre study of pediatric HCAIs caused by S.aureus from a tertiary care hospital in Turkey between February 2014 and December 2019.The clinical and epidemiological characteristics and antimicrobial susceptibility of the methicillin-susceptible and methicillin-resistant S.aureus(MSSA and MRSA)isolates was evaluated.Results:A total of 310 pediatric patients were examined.Overall,225(72.6%)isolates were MSSA and 85(27.4%)were MRSA.All S.aureus isolates were susceptible to teicoplanin,vancomycin,linezolid,tigecycline,mupirocin,and daptomycin.Penicillin resistance rates were high(89.0%),while fosfomycin,gentamicin,and clindamycin resistance rates were low(1.3%,1.0%,and 2.3%,respectively).Except susceptibility to fosfomycin,which was significantly lower in 2014 compared to 2018 and 2019,no significant difference was found in the antimicrobial susceptibility of S.aureus isolates between the years.Baseline characteristics and mortality rate were similar comparing MRSA and MSSA causing HCAIs.The mortality rate of HCAIs caused by S.aureus was 6.5%(20 patients).Malignancy was an independent risk factor associated with mortality in the multivariate analysis(OR 5.446,95%CI 1.573-18.849).Conclusions:Our findings demonstrate that MSSA remained the most causative agent of HCAIs caused by S.aureus.The mortality rate was 6.5%,the antibiotic resistance rate was quite high for penicillin and diagnosis of malignancy was the main risk factor for increasing mortality in children.These findings could help improve the management of HCAIs caused by S.aureus in children.展开更多
BACKGROUND Although neonatal Staphylococcus aureus pneumonia is common and usually curable,it can also be refractory and life-threatening.Herein,we report a case of severe neonatal community-acquired methicillin-resis...BACKGROUND Although neonatal Staphylococcus aureus pneumonia is common and usually curable,it can also be refractory and life-threatening.Herein,we report a case of severe neonatal community-acquired methicillin-resistant Staphylococcus aureus(CA-MRSA)necrotizing pneumonia with bilateral recurrent pyopneumothorax,respiratory failure,heart failure,and cardiac arrest.We hope our report will add to the understanding of this disease.CASE SUMMARY An 18-d-old boy presented with cough for five days,fever for three days,and dyspnea for two days.Preadmission chest radiograph revealed high-density shadows in both lungs.On admission,his oxygen saturation fluctuated around 90%under synchronized intermittent mandatory ventilation.He was unconscious,with dyspnea,weak heart sounds and hepatomegaly.Moist crackles were present throughout his left lung,while the breath sounds in the right lung were decreased.After high-frequency oscillatory ventilation,empiric antimicrobials(meropenem and vancomycin),improved circulation,and right pleural cavity drainage for right pneumothorax(approximately 90%compression),his oxygen saturation level stayed above 95%,and recruitment of the right lung was observed.His condition did not deteriorate until the 5th day of hospitalization(DOH 5).On the morning of DOH 5,his oxygen saturation decreased.Subsequent chest radiograph showed bilateral pneumothorax with nearly 100%compression of the left lung.Desaturation was not relieved after urgent left pleural cavity drainage,and cardiac arrest occurred soon thereafter.Although his spontaneous heartbeat returned through emergency resuscitation and salvage antibacterial therapy(linezolid and levofloxacin)was administered given the detection and antimicrobial susceptibility of MRSA,he showed no improvement,with recurrent pyopneumothorax and continued drainage of purulent fluid and necrotic lung tissue fragments from the pleural cavity.Eventually,his parents refused extracorporeal membrane oxygenation(ECMO)and gave up all the treatments,and the newborn passed away soon after withdrawal on DOH 13.CONCLUSION Neonatal MRSA pneumonia can be refractory and lethal,especially in cases where necrotizing pneumonia leads to extensive lung necrosis and recurrent pneumothorax.Despite treatment with linezolid and other medical measures,it may still be ineffective.Currently,ECMO has been a remedial therapy,but if the lung tissue is too severely eroded to be repaired,it may be useless unless the infection can be controlled and lung transplantation can be performed.Regardless of whether ECMO is initiated,the key to successful treatment is to achieve control over the pneumonia caused by MRSA as soon as possible and to reverse lung injury as much as possible.展开更多
[Objectives]To observe the effects of daphnetin on mastitis induced by Staphylococcus aureus in mice.[Methods]18 postpartum ICR female mice were used to establish mastitis animal model,and were randomly divided into t...[Objectives]To observe the effects of daphnetin on mastitis induced by Staphylococcus aureus in mice.[Methods]18 postpartum ICR female mice were used to establish mastitis animal model,and were randomly divided into three groups(A,B,and C)with 6 mice in each group.Group A:blank control group;group B:S.Aureus model group;group C:S.Aureus model+daphnetin group.The experimental groups were injected 1 mL of 1.0×104 CFU/100μL of S.aureus of along the nipple catheter.The suspension was placed in the 3 rd and 4 th pairs of mammary glands,and the control group was injected with the same dose of normal saline.On the second day after infection,the rats in group A,B and C were given drugs by gavage,while the rats in group A and B were given normal saline and the rats in group C were given daphnetin once a day for 6 consecutive days.Blood samples were collected from living eyeballs,and blood cells were analyzed by automatic flow cytometer after anticoagulation.[Results]The NLR and Systemie Immune Inflammati-on Index(SII)in the blood of mastitis mice induced by S.aureus were significantly higher than those in the control group(P<0.01),suggesting that neutrophil to lymphocyte ratio(NLR)and SII can be used as diagnostic indicators of mastitis,and the levels of NLR and SII decreased significantly after daphnetin intervention.[Conclusions]NLR and SII showed high levels in mastitis mice,which are valuable for the diagnosis of mastitis and the evaluation of its prognosis.After the intervention of daphnetin,both of them decreased significantly,indicating that daphnetin has a good prognosis trend in mastitis mice induced by S.aureus.展开更多
Persistent methicillin-sensitive staphylococcus aureus(MSSA)bacteremia is usually defined as persistent bacteremia despite 7 days of appropriate antibiotic therapy.While nafcillin,cefazolin,or oxacillin are excellent ...Persistent methicillin-sensitive staphylococcus aureus(MSSA)bacteremia is usually defined as persistent bacteremia despite 7 days of appropriate antibiotic therapy.While nafcillin,cefazolin,or oxacillin are excellent antimicrobial options,persistent MSSA is a commonly encountered phenomenon in clinical practice for which further guidelines on appropriate management are necessitated.Here we highlight two cases of MSSA bacteremia requiring salvage therapy with ertapenem.展开更多
Staphylococcus aureus is a gram-staining positive cocci bacillus baterium and also one of the foodborne pathogens, which is a serious potential hazard to human health and food safety. We constructed an electroche...Staphylococcus aureus is a gram-staining positive cocci bacillus baterium and also one of the foodborne pathogens, which is a serious potential hazard to human health and food safety. We constructed an electrochemical biosensor for the detection of S. aureus based on nucleic acid aptamers to achieve highly specific detection of S. aureus. The detection of S. aureus was realized by using Aptamer (Apt) to capture S. aureus, which resulted in a change in the spatial conformation of Apt and a decrease in the electrochemical signal. Under the optimized experimental conditions, the detected electrochemical signals were positively correlated with the concentration of S. aureus with a linear range of 1 × 10<sup>1</sup> - 1 × 10<sup>5</sup> CFU/mL, a detection limit of 4.76 CFU/mL, and an experimental recovery of 97.43% - 99.37%. Therefore, we successfully constructed an electrochemical biosensor for the specific detection of S. aureus, which has the advantages of high specificity, sensitive detection and convenient operation.展开更多
Objective: To investigate the effects of Andrographis paniculata(Burm.f.) Wall. Ex Nees(A. paniculata) on expressions and activities of catalase, superoxide dismutase and alkylhydroperoxide reductase C in Staphylococc...Objective: To investigate the effects of Andrographis paniculata(Burm.f.) Wall. Ex Nees(A. paniculata) on expressions and activities of catalase, superoxide dismutase and alkylhydroperoxide reductase C in Staphylococcus aureus(S. aureus) with respect to its survival in vitro. Methods: Antioxidative property of methanolic leaves extract of A. paniculata(0.06 mg/m L). Minimum inhibitory concentration(MIC) was determined by its ability to reduce hydrogen peroxide(H_2O_2) toxicity against S. aureus ATCC 25923(3.8×10~8 cfu/m L). Effects of the extract on expressions of kat A(encoding catalase), sod A and sod M [encoding superoxide dismutases(SODs)], and ahp C [encoding alkylhydroperoxide reductase C(AhpC)] in S. aureuswere determined by RT-q PCR and corresponding enzyme activity assays were performed. Nitroblue tetrazolium reduction(NBT) assay was performed to determine effects of the extract on intracellular and extracellular levels of O_2-in S. aureus. Results: Cells challenged with 7.5 mmol/L H_2O_2 showed 0% survival in 30 min whereas 25% survived after treatment with the extract and H_2O_2. Cells that were treated with the extract alone had 43% survival in the same exposure period. Expressions of sod A and sod M genes in extract-treated cells were lowered 0.8-fold and 0.7-fold, respectively with decrease in total SOD activity of 26.8 U compared to untreated cells, 32.4 U(P<0.05). In contrast, extract-treated S. aureus cells showed 3.3-fold increase in kat A expression with corresponding increase in catalase activity of 1.828 U compared to untreated cells which was 1.248 U,(P<0.05). More profoundly, ahp C expression was increased 61-fold in extract-treated cells,(P<0.05) with corresponding increase in Ahp C activity of 0.018 U compared to untreated cells, 0.012U,(P<0.05). Extract-treated cells had significantly lower intra-and extracellular O_2-levels with absorbance readings(A_(575nm)) of 0.340 and 0.524 compared to untreated cells which were 0.516 and 0.928(P<0.05), respectively. Conclusions: Taken together these results suggest that the low MIC of A. paniculatamethanolic leaves extract(0.06 mg/m L) reduce H_2O_2 toxicity and more importantly, was in itself effectively inhibitory against S. aureus. Further, our observations suggest that a probable mode of its inhibitory mechanism against S. aureus is by reducing total SOD activity through downregulation of sod A and sod M expressions.展开更多
Objective: To isolate a-mangostin(AMG) from the peels of mangosteen(Garcinia mangostana L.), grown in Vietnam, and to investigate antibiofilm activity of this compound against three Staphylococcus aureus(S. aureus) st...Objective: To isolate a-mangostin(AMG) from the peels of mangosteen(Garcinia mangostana L.), grown in Vietnam, and to investigate antibiofilm activity of this compound against three Staphylococcus aureus(S. aureus) strains, one of which was methicillin-resistant S. aureus(MRSA) and the other two strains were methicillinsensitive S. aureus(MSSA).Methods: AMG in n-hexane fraction was isolated on a silica gel column and chemically analyzed by HPLC and NMR. The antibiofilm activity of this compound was investigated by using a 96-well plate model for the formation of biofilms. Biofilm biomass was quantified using crystal violet. The viability of cells was observed under confocal microscopy using LIVE/DEAD Bac Light stains. Biofilm composition was determined using specific chemical and enzyme tests for polysaccharide, protein and DNA. Membranedamaging activity was assayed by measuring the hemolysis of human red blood cells in presence of AMG.Results: The results indicated that the isolated AMG, with a purity that exceeded 98%,had minimal inhibitory concentrations in the range of 4.6–9.2 mmol/L for the three strains tested. Interestingly, the MSSA strains were more sensitive to AMG than the MRSA strain. Minimal bactericidal concentrations were 2-fold higher than the minimal inhibitory concentration values for the three strains, indicating that AMG was a bactericidal compound. AMG also prevented biofilm formation effectively, albeit that again the MRSA strain was the most resistant. Interestingly, biofilms of the MRSA strain contained protein as a main component of the extracellular matrix, whereas this was polysaccharide in the MSSA strains. This might relate to the resistance of the MRSA 252 strain to AMG.Assays using human red blood cells indicated that AMG caused significant membrane damage with 50% of cell lysis occurred at concentration of about 36 mmol/L.Conclusions: Our results provide evidence that the isolated AMG has inhibitory activity against biofilm formation by S. aureus, including MRSA. Thus, isolated AMG proposes a high potential to develop a novel phytopharmaceutical for the treatment of MRSA.展开更多
Ultrasound,is thought to a potential non-thermal sterilization technology in food industry.However,the exact mechanisms underlying microbial inactivation by ultrasound still remain obscure.In this study,the action mod...Ultrasound,is thought to a potential non-thermal sterilization technology in food industry.However,the exact mechanisms underlying microbial inactivation by ultrasound still remain obscure.In this study,the action modes of ultrasound on both Gram-negative and Gram-positive microorganisms were estimated.From colony results,ultrasound acted as an irreversible effect on both Eshcerichia coli and Staphylococcus aureus without sublethal injury.The result in this study also showed that a proportion of bacteria subpopulation suffered from serious damage of intracellular components(e.g.DNA and enzymes)but with intact cell envelopes.We speculated that the inactivated effects of ultrasound on microbes might more than simply completed disruption of cell exteriors.Those microbial cells who had not enter the valid area of ultrasonic cavitation might be injected with free radicals produced by ultrasound and experienced interior injury with intact exterior structure,and others who were in close proximity to the ultrasonic wave field would be immediately and completely disrupted into debris by high power mechanic forces.These findings here try to provide extension for the inactivation mechanisms of ultrasound on microorganisms.展开更多
Cefquinome is the only fourth-generation cephalosporin used solely for veterinary applications.In this study,we established the wild-type cut-off(CO_(WT))and pharmacokinetic/pharmacodynamic cut-off(CO_(PD))of cefquino...Cefquinome is the only fourth-generation cephalosporin used solely for veterinary applications.In this study,we established the wild-type cut-off(CO_(WT))and pharmacokinetic/pharmacodynamic cut-off(CO_(PD))of cefquinome against Escherichia coli and Staphylococcus aureus.A total of 210 E.coli and 160 S.aureus isolates were collected from pigs in Guangdong Province between 2014 and 2018.The minimum inhibitory concentrations(MICs)were determined using a microdilution broth method.MIC_(50)and MIC_(90)were 0.06 and 0.25μg m L^(–1)for E.coli and 0.5 and 1μg m L^(–1)for S.aureus,respectively.Statistical analysis and the ECOFFinder Program showed that the CO_(WT)for cefquinome against E.coli and S.aureus were0.125 and 2μg m L^(–1),respectively.The resistance rates were 11.9%for E.coli and 6.25%for S.aureus.Based on a5000-subject Monte Carlo simulation,the CO_(PD)value for cefquinome against E.coil and S.aureus was 0.25μg m L^(–1)under the recommended dose(2 mg kg^(–1),twice a day for 3 days),confirming that infections caused by strains with MIC≤0.25μg m L^(–1)could be effectively treated.Following adjustment of the dosing regimen to 4.5 mg kg^(–1),effective treatment(>90)was achieved for S.aureus infections with MIC_(90)1μg m L^(–1).This susceptibility breakpoint determination is significant for resistant surveillance and cefquinome dosage guidance against E.coli and S.aureus in pigs.展开更多
Background: Mastitis in dairy cows caused by Staphylococcus aureus is a major problem hindering economic growth in dairy farms worldwide. It is difficult to prevent or eliminate due to its asymptomatic nature and long...Background: Mastitis in dairy cows caused by Staphylococcus aureus is a major problem hindering economic growth in dairy farms worldwide. It is difficult to prevent or eliminate due to its asymptomatic nature and long persistence of infection. Although transcriptomic responses of bovine mammary gland cells to pathogens that cause mastitis have been studied, the common responses of peripheral blood leukocytes to S. aureus infection across two consecutive generations of dairy cattle have not been investigated.Methods: In the current study, RNA-Seq was used to profile the transcriptomes of peripheral blood leukocytes sampled from S. aureus-infected mothers and their S. aureus-infected daughters, and also healthy non-infected mothers and their healthy daughters. Differential gene expression was evaluated as follows: 1) S. aureus-infected cows versus healthy non-infected cows(S vs. H, which include all the mothers and daughters), 2) S. aureus-infected mothers versus healthy non-infected mothers(SM vs. HM), and 3) S. aureus-infected daughters versus healthy noninfected daughters(SMD vs. HMD).Results: Analysis of all identified expressed genes in the four groups(SM, SMD, HM, and HMD) showed that EPOR,IL9, IFNL3, CCL26, IL26 were exclusively expressed in both the HM and HMD groups, and that they were significantly(P < 0.05) enriched for the cytokine-cytokine receptor interaction pathway. A total of 17, 13 and 10 differentially expressed genes(DEGs)(FDR Padj. < 0.1 and |FC| > 1.2) were detected in the three comparisons, respectively. DEGs with P < 0.05 and |FC| > 2 were used for functional enrichment analyses. For the S vs. H comparison, DEGs detected included CCL20, IL13 and MMP3, which are associated with the IL-17 signaling pathway. In the SM vs. HM and SMD vs. HMD comparisons, five(BLA-DQB, C1 R, C2, FCGR1 A, and KRT10) and six(BLA-DQB, C3 AR1, CFI, FCAR, FCGR3 A, and LOC10498484) genes, respectively, were involved in the S. aureus infection pathway.Conclusions: Our study provides insights into the transcriptomic responses of bovine peripheral blood leukocytes across two generations of cattle naturally infected with S. aureus. The genes highlighted in this study could serve as expression biomarkers for mastitis and may also contain sequence variation that can be used for genetic improvement of dairy cattle for resilience to mastitis.展开更多
Background:Staphylococcus aureus(S.aureus)mastitis is one of the most difficult diseases to treat in lactating dairy cows worldwide.S.aureus with different lineages leads to different host immune responses.Long non-co...Background:Staphylococcus aureus(S.aureus)mastitis is one of the most difficult diseases to treat in lactating dairy cows worldwide.S.aureus with different lineages leads to different host immune responses.Long non-coding RNAs(lncRNAs)are reported to be widely involved in the progress of inflammation.However,no research has identified stable lncRNAs among different S.aureus strain infections.In addition,folic acid(FA)can effectively reduce inflammation,and whether the inflammatory response caused by S.aureus can be reduced by FA remains to be explored.Methods:lncRNA transcripts were identified from Holstein mammary gland tissues infected with different concentrations of S.aureus(in vivo)and mammary alveolar cells(Mac-T cells,in vitro)challenged with different S.aureus strains.Differentially expressed(DE)lncRNAs were evaluated,and stable DE lncRNAs were identified in vivo and in vitro.On the basis of the gene sequence conservation and function conservation across species,key lncRNAs with the function of potentially immune regulation were retained for further analysis.The function of FA on inflammation induced by S.aureus challenge was also investigated.Then,the association analysis between these keys lncRNA transcripts and hematological parameters(HPs)was carried out.Lastly,the knockdown and overexpression of the important lncRNA were performed to validate the gene function on the regulation of cell immune response.Results:Linear regression analysis showed a significant correlation between the expression levels of lncRNA shared by mammary tissue and Mac-T cells(P<0.001,R^(2)=0.3517).lncRNAs PRANCR and TNK2-AS1 could be regarded as stable markers associated with bovine S.aureus mastitis.Several HPs could be influenced by SNPs around lncRNAs PRANCR and TNK2-AS1.The results of gene function validation showed PRANCR regulates the mRNA expression of SELPLG and ITGB2 within the S.aureus infection pathway and the Mac-T cells apoptosis.In addition,FA regulated the expression change of DE lncRNA involved in toxin metabolism and inflammation to fight against S.aureus infection.Conclusions:The remarkable association between SNPs around these two lncRNAs and partial HP indicates the potentially important role of PRANCR and TNK2-AS1 in immune regulation.Stable DE lncRNAs PRANCR and TNK2-AS1 can be regarded as potential targets for the prevention of bovine S.aureus mastitis.FA supplementation can reduce the negative effect of S.aureus challenge by regulating the expression of lncRNAs.展开更多
Staphylococcus aureus(S.aureus)is a leading human pathogen capable of producing severe invasive infections such as bacteremia,sepsis,and endocarditis with high morbidity and mortality,exacerbated by the increasingly w...Staphylococcus aureus(S.aureus)is a leading human pathogen capable of producing severe invasive infections such as bacteremia,sepsis,and endocarditis with high morbidity and mortality,exacerbated by the increasingly widespread antibiotic resistance exemplified by methicillin-resistant strains(MRSA).S.aureus pathogenesis is fueled by the secretion of toxins—such as the membrane-damaging pore-forming atoxin,which have diverse cellular targets including the epithelium,endothelium,leukocytes,and platelets.Here,we examine the use of human platelet membrane-coated nanoparticles(PNPs)as a biomimetic decoy strategy to neutralize S.aureus toxins and preserve host cell defense functions.The PNPs blocked platelet damage induced by S.aureus secreted toxins,thereby supporting platelet activation and bactericidal activity.Likewise,the PNPs blocked macrophage damage induced by S.aureus secreted toxins,thus supporting macrophage oxidative burst,nitric oxide production,and bactericidal activity,and diminishing MRSA-induced neutrophil extracellular trap release.In a mouse model of MRSA systemic infection,PNP administration reduced bacterial counts in the blood and protected against mortality.Taken together,the results from the present work provide a proof of principle of the therapeutic benefit of PNPs in toxin neutralization,cytoprotection,and increased host resistance to invasive S.aureus infection.展开更多
基金supported by Brain Korea (BK)21 Plus Project (4299990913942)funded by the Korean Government,Koreathe Collabo Project funded by the Ministry of SMEs and Startups (C1016120-01-02)the National Research Foundation of Korea (NRF) (2018007551)。
文摘Slightly acidic electrolyzed water(SAEW)has proven to be an efficient and novel sanitizer in food and agriculture field.This study assessed the efficacy of SAEW(30 mg/L)at 40℃on the inactivation of foodbome pathogens and detachment of multi-resistant Staphylococcus aureus(MRSA)biofilm.Furthermore.the underlying mechanism of MRS A biofilm under heated SAEW at 40℃treatment on metabolic profiles was investigated.The results showed that the heated SAEW at 40℃significantly effectively against foodbome pathogens of 1.96-7.56(lg(CFU/g))reduction in pork,chicken,spinach,and lettuce.The heated SAEW at 40℃treatment significantly reduced MRS A biofilm cells by 2.41(lg(CFU/cm^(2))).The synergistic effect of SAEW treatment showed intense anti-biofilm activity in decreasing cell density and impairing biofilm cell membranes.Global metabolic response of MRSA biofilms,treated by SAEW at 40℃,revealed the alterations of intracellular metabolites,including amino acids,organic acid,fatty acid,and lipid.Moreover,signaling pathways involved in amino acid metabolism,energy metabolism,nucleotide synthesis,carbohydrate metabolites,and lipid biosynthesis were functionally disrupted by the SAEW at 40℃treatment.As per our knowledge,this is the first research to uncover the potential mechanism of heated SAEW treatment against MRSA biofilm on food contact surface.
基金supported by the National Natural Science Foundation of China (31972169 and 32001798)。
文摘Staphylococcus aureus is a serious foodborne pathogen threatening food safety and public health.Especially the emergence of methicillin-resistant Staphylococcus aureus(MRSA)increased the difficulty of S.aureus treatment.Staphyloxanthin is a crucial virulence factor of S.aureus.Blocking staphyloxanthin production could help the host immune system counteract the invading S.aureus cells.In this study,we first screened for staphyloxanthin inhibitors using a virtual screening method.The outcome of the virtual screening method resulted in the identification of eugenol(300μg/mL),which significantly inhibits the staphyloxanthin production in S.aureus ATCC 29213,S.aureus Newman,MRSA ATCC 43300 and MRSA ATCC BAA1717by 84.2%,63.5%,68.1%,and 79.5%,respectively.The outcome of the growth curve assay,field-emission scanning electron,and confocal laser scanning microscopy analyses confirmed that eugenol at the test concentration did not affect the morphology and growth of S.aureus.Moreover,the survival rate of S.aureus ATCC 29213 and MRSA ATCC 43300 under H_(2)O_(2) pressure decreased to 51.9%and 45.5%in the presence of eugenol,respectively.The quantitative RT-PCR and molecular simulation studies revealed that eugenol targets staphyloxanthin biosynthesis by downregulating the transcription of the crtM gene and inhibiting the activity of the CrtM enzyme.Taken together,we first determined that eugenol was a prominent compound for staphyloxanthin inhibitor to combat S.aureus especially MRSA infections.
文摘Introduction: Bacterial skin and soft tissue infections (SSTIs) are a cause of frequent inpatient and outpatient care visits whose causative agents are associated with a high antimicrobial resistance burden. For insights on antimicrobial susceptibilities in a rural setting, we examined specimens from suspected SSTIs from two public health facilities in Kenya. We additionally assessed antibiotic use, appropriateness of empiric therapy and risk factors for SSTI. Methodology: Between 2021 and 2023, 265 patients at Kisii and Nyamira County Referral hospitals were enrolled. Wound swabs/aspirates were collected and processed following standard microbiological procedures. Identification and antimicrobial susceptibility were performed using the VITEK 2 Compact platform. Demographic, clinical, and microbiological data were analyzed with R Statistical software. Results: S. aureus was isolated in 16.2% (43/265) of patients with a methicillin resistance (MRSA) proportion of 14% (6/43). While 13/15 drugs elicited susceptibilities ranging from 84% - 100%, penicillin (16%) and trimethoprim-sulfamethoxazole [TMP-SXT] (23%) yielded the lowest susceptibilities. Escherichia coli (n = 33), Klebsiella pneumoniae (n = 8), Pseudomonas aeruginosa (n = 8), and Citrobacter species (n = 4) were the most commonly isolated gram-negative species. Gram-negative strains showed high susceptibilities to most of the tested drugs (71% - 100%) with the exception of ampicillin (18%), TMP-SXT (33%), and first and second generation cephalosporins. Conclusions: The low MRSA prevalence and generally high antibiotic susceptibilities for S. aureus and gram-negative bacteria present opportunities for antibiotic stewardship in the study setting. Diminished susceptibilities against penicillin/ampicillin and TMP-SXT accord with prevailing local data and add a layer of evidence for their cautious empiric use.
基金The study was funded by the UPNM Short Term Grant (UPNM/2019/GPJP/SP/1).
文摘Objective:To determine the antibiotic resistance profile(ARP)of Staphylococcus(S.)aureus isolates and molecular typing of the methicillin-resistant S.aureus(MRSA)isolates from Tuanku Mizan Armed Forces Hospital(TMAFH),Kuala Lumpur.Methods:The ARP and presence of the pvl gene were determined for 209 S.aureus isolates from clinical specimens.Of these,123 were methicillin-susceptible S.aureus(MSSA)isolates and 86 were MRSA isolates.All MRSA isolates were characterized using SCCmec typing and spa typing.Descriptive analysis was performed to compare the demographic data with the phenotypic and genotypic variables of the S.aureus isolates.Results:No vancomycin-intermediate and-resistant S.aureus(VISA and VRSA,respectively)were detected among the study isolates.The MSSA isolates showed low resistance rates to all tested antibiotics,were commonly invasive(28/42,66.7%),and mostly harboured pvl(35/42,83.3%).Meanwhile,MRSA isolates showed high resistance to penicillin(86/86,100%),ampicillin(86/86,100%),sulbactam/ampicillin(86/86,100%),cefuroxime(81/86,94.19%),cefoperazone(76/86,88.37%),azithromycin(56/86,65.12%),and erythromycin(54/86,62.79%).The majority of MRSA isolates were of SCCmec type IVh(65/86,75.58%),spa type t032(55/85,63.95%),and grouped into spaCC-t022(66/85,77.65%).The t032 type was found to be associated with resistance traits to azithromycin and erythromycin(P<0.05).We also found several spa types that are typically associated with hospital-,community-,and livestock-associated MRSA co-existing in our MRSA population.Conclusions:This study reflected the consistent absence of VISA and VRSA and corroborated the clonal shifting of MRSA isolates in the Malaysian MRSA isolates.
基金supported by the National Natural Science Foundation of China(U22A20523,32172912,and 32102722)the Interdisciplinary Integration and Innovation Project of Jilin University(JLUXKJC2021QZ04)。
文摘Host-directed therapy(HDT)is an emerging novel approach for treating multidrug-resistant Staphylococcus aureus(S.aureus)infection.Functioning as the indispensable specific cellular receptor for a-toxin(Hla),a-disintegrin and metalloproteinase 10(ADAM10)is exploited to accelerate S.aureus infection through diverse mechanisms.The extraordinary contribution of ADAM10 to S.aureus pathogenesis renders it an attractive HDT target for combating S.aureus infection.Our study is the first to demonstrate the indispensable role of ADAM10 in S.aureus-induced necroptosis,and it enhances our knowledge of the role of ADAM10 in S.aureus infection.Using a fluorogenic substrate assay,we further identified kaempferol as a potent ADAM10 inhibitor that effectively protected mice from S.aureus infection by suppressing Hla-mediated barrier disruption and necroptosis.Collectively,our work presents a novel hostdirected therapeutic strategy for using the promising candidate kaempferol to treat S.aureus infection and other diseases relevant to the disordered upregulation of ADAM10.
基金National Science and Technology Major Project,Grant/Award Number:2016YFD0500208,2017ZX10304402-001-012 and 2017ZX10304402-001-006Shanghai Science and Technology Commission“R&D public service platform and institutional capacity improvement project”,Grant/Award Number:21DZ2291300Shanghai Public Health Clinical Center projects,Grant/Award Number:KY-GW-2021-39,KY-GW-2019-19 and KY-GW-2019-11。
文摘Background:Staphylococcus aureus can cause serious infections by secreting many superantigen exotoxins in“carrier”or“pathogenic”states.HLA DQ and HLA DR humanized mice have been used as a small animal model to study the role of two molecules during S.aureus infection.However,the contribution of HLA DP to S.aureus infection is unknown yet.Methods:In this study,we have produced HLA DP401 and HLA DRA0101 humanized mice by microinjection of C57BL/6J zygotes.Neo-floxed IAβ+/-mice were crossbred with Ella-Cre and further crossbred with HLA DP401 or HLA-DRA0101 humanized mice.After several rounds of traditional crossbreeding,we finally obtained HLA DP401-IAβ-/-and HLA DRA-IAβ-/-humanized mice,in which human DP401 or DRA0101 molecule was introduced into IAβ-/-mice deficient in endogenous murine MHC classⅡmolecules.A transnasal infection murine model of S.aureus pneumonia was induced in the humanized mice by administering 2×108CFU of S.aureus Newman dropwise into the nasal cavity.The immune responses and histopathology changes were further assessed in lungs in these infected mice.Results:We evaluated the local and systemic effects of S.aureus delivered intranasally in HLA DP401-IAβ-/-and HLA DRA-IAβ-/-transgenic mice.S.aureus Newman infection significantly increased the m RNA level of IL 12p40 in lungs in humanized mice.An increase in IFN-γand IL-6 protein was observed in HLA DRA-IAβ-/-mice.We observed a declining trend in the percentage of F4/80+macrophages in lungs in HLA DP401-IAβ-/-mice and a decreasing ratio of CD4+to CD8+T cells in lungs in IAβ-/-mice and HLA DP401-IAβ-/-mice.A decreasing ratio of Vβ3+to Vβ8+T cells was also found in the lymph node of IAβ-/-mice and HLA DP401-IAβ-/-mice.S.aureus Newman infection resulted in a weaker pathological injury in lungs in IAβ-/-genetic background mice.Conclusion:These humanized mice will be an invaluable mouse model to resolve the pathological mechanism of S.aureus pneumonia and study what role DP molecule plays in S.aureus infection.
文摘Objective:To describe clinical and epidemiological characteristics,antimicrobial susceptibility and mortality-associated factors of healthcare-associated infections(HCAIs)caused by Staphylococcus(S.)aureus in children.Methods:We conducted a retrospective,single-centre study of pediatric HCAIs caused by S.aureus from a tertiary care hospital in Turkey between February 2014 and December 2019.The clinical and epidemiological characteristics and antimicrobial susceptibility of the methicillin-susceptible and methicillin-resistant S.aureus(MSSA and MRSA)isolates was evaluated.Results:A total of 310 pediatric patients were examined.Overall,225(72.6%)isolates were MSSA and 85(27.4%)were MRSA.All S.aureus isolates were susceptible to teicoplanin,vancomycin,linezolid,tigecycline,mupirocin,and daptomycin.Penicillin resistance rates were high(89.0%),while fosfomycin,gentamicin,and clindamycin resistance rates were low(1.3%,1.0%,and 2.3%,respectively).Except susceptibility to fosfomycin,which was significantly lower in 2014 compared to 2018 and 2019,no significant difference was found in the antimicrobial susceptibility of S.aureus isolates between the years.Baseline characteristics and mortality rate were similar comparing MRSA and MSSA causing HCAIs.The mortality rate of HCAIs caused by S.aureus was 6.5%(20 patients).Malignancy was an independent risk factor associated with mortality in the multivariate analysis(OR 5.446,95%CI 1.573-18.849).Conclusions:Our findings demonstrate that MSSA remained the most causative agent of HCAIs caused by S.aureus.The mortality rate was 6.5%,the antibiotic resistance rate was quite high for penicillin and diagnosis of malignancy was the main risk factor for increasing mortality in children.These findings could help improve the management of HCAIs caused by S.aureus in children.
基金Supported by the Scientific and Technological Project of Shiyan City of Hubei Province,No.21Y32the Medical Research Project of Hubei Pediatric Alliance,No.HPAMRP202114.
文摘BACKGROUND Although neonatal Staphylococcus aureus pneumonia is common and usually curable,it can also be refractory and life-threatening.Herein,we report a case of severe neonatal community-acquired methicillin-resistant Staphylococcus aureus(CA-MRSA)necrotizing pneumonia with bilateral recurrent pyopneumothorax,respiratory failure,heart failure,and cardiac arrest.We hope our report will add to the understanding of this disease.CASE SUMMARY An 18-d-old boy presented with cough for five days,fever for three days,and dyspnea for two days.Preadmission chest radiograph revealed high-density shadows in both lungs.On admission,his oxygen saturation fluctuated around 90%under synchronized intermittent mandatory ventilation.He was unconscious,with dyspnea,weak heart sounds and hepatomegaly.Moist crackles were present throughout his left lung,while the breath sounds in the right lung were decreased.After high-frequency oscillatory ventilation,empiric antimicrobials(meropenem and vancomycin),improved circulation,and right pleural cavity drainage for right pneumothorax(approximately 90%compression),his oxygen saturation level stayed above 95%,and recruitment of the right lung was observed.His condition did not deteriorate until the 5th day of hospitalization(DOH 5).On the morning of DOH 5,his oxygen saturation decreased.Subsequent chest radiograph showed bilateral pneumothorax with nearly 100%compression of the left lung.Desaturation was not relieved after urgent left pleural cavity drainage,and cardiac arrest occurred soon thereafter.Although his spontaneous heartbeat returned through emergency resuscitation and salvage antibacterial therapy(linezolid and levofloxacin)was administered given the detection and antimicrobial susceptibility of MRSA,he showed no improvement,with recurrent pyopneumothorax and continued drainage of purulent fluid and necrotic lung tissue fragments from the pleural cavity.Eventually,his parents refused extracorporeal membrane oxygenation(ECMO)and gave up all the treatments,and the newborn passed away soon after withdrawal on DOH 13.CONCLUSION Neonatal MRSA pneumonia can be refractory and lethal,especially in cases where necrotizing pneumonia leads to extensive lung necrosis and recurrent pneumothorax.Despite treatment with linezolid and other medical measures,it may still be ineffective.Currently,ECMO has been a remedial therapy,but if the lung tissue is too severely eroded to be repaired,it may be useless unless the infection can be controlled and lung transplantation can be performed.Regardless of whether ECMO is initiated,the key to successful treatment is to achieve control over the pneumonia caused by MRSA as soon as possible and to reverse lung injury as much as possible.
基金Supported by Project of Innovation and Entrepreneurship Training Program for College Students in Guangxi Zhuang Autonomous Region in 2022(S202210599060)Basic Ability Improvement Project for Young and Middle-aged Teachers in Guangxi Zhuang Autonomous Region in 2020(2020KY13027)Project of Scientific Research and Technological Development Plan of Baise City in 2020(Encyclopedia[2020]No.47).
文摘[Objectives]To observe the effects of daphnetin on mastitis induced by Staphylococcus aureus in mice.[Methods]18 postpartum ICR female mice were used to establish mastitis animal model,and were randomly divided into three groups(A,B,and C)with 6 mice in each group.Group A:blank control group;group B:S.Aureus model group;group C:S.Aureus model+daphnetin group.The experimental groups were injected 1 mL of 1.0×104 CFU/100μL of S.aureus of along the nipple catheter.The suspension was placed in the 3 rd and 4 th pairs of mammary glands,and the control group was injected with the same dose of normal saline.On the second day after infection,the rats in group A,B and C were given drugs by gavage,while the rats in group A and B were given normal saline and the rats in group C were given daphnetin once a day for 6 consecutive days.Blood samples were collected from living eyeballs,and blood cells were analyzed by automatic flow cytometer after anticoagulation.[Results]The NLR and Systemie Immune Inflammati-on Index(SII)in the blood of mastitis mice induced by S.aureus were significantly higher than those in the control group(P<0.01),suggesting that neutrophil to lymphocyte ratio(NLR)and SII can be used as diagnostic indicators of mastitis,and the levels of NLR and SII decreased significantly after daphnetin intervention.[Conclusions]NLR and SII showed high levels in mastitis mice,which are valuable for the diagnosis of mastitis and the evaluation of its prognosis.After the intervention of daphnetin,both of them decreased significantly,indicating that daphnetin has a good prognosis trend in mastitis mice induced by S.aureus.
文摘Persistent methicillin-sensitive staphylococcus aureus(MSSA)bacteremia is usually defined as persistent bacteremia despite 7 days of appropriate antibiotic therapy.While nafcillin,cefazolin,or oxacillin are excellent antimicrobial options,persistent MSSA is a commonly encountered phenomenon in clinical practice for which further guidelines on appropriate management are necessitated.Here we highlight two cases of MSSA bacteremia requiring salvage therapy with ertapenem.
文摘Staphylococcus aureus is a gram-staining positive cocci bacillus baterium and also one of the foodborne pathogens, which is a serious potential hazard to human health and food safety. We constructed an electrochemical biosensor for the detection of S. aureus based on nucleic acid aptamers to achieve highly specific detection of S. aureus. The detection of S. aureus was realized by using Aptamer (Apt) to capture S. aureus, which resulted in a change in the spatial conformation of Apt and a decrease in the electrochemical signal. Under the optimized experimental conditions, the detected electrochemical signals were positively correlated with the concentration of S. aureus with a linear range of 1 × 10<sup>1</sup> - 1 × 10<sup>5</sup> CFU/mL, a detection limit of 4.76 CFU/mL, and an experimental recovery of 97.43% - 99.37%. Therefore, we successfully constructed an electrochemical biosensor for the specific detection of S. aureus, which has the advantages of high specificity, sensitive detection and convenient operation.
基金supported by the Faculty of Health Sciences,Universiti Teknologi MARA,Puncak Alam Campus,Selangor and LESTARI grant(600-IRMI/MYRA 5/3/LESTARI(16/2016)
文摘Objective: To investigate the effects of Andrographis paniculata(Burm.f.) Wall. Ex Nees(A. paniculata) on expressions and activities of catalase, superoxide dismutase and alkylhydroperoxide reductase C in Staphylococcus aureus(S. aureus) with respect to its survival in vitro. Methods: Antioxidative property of methanolic leaves extract of A. paniculata(0.06 mg/m L). Minimum inhibitory concentration(MIC) was determined by its ability to reduce hydrogen peroxide(H_2O_2) toxicity against S. aureus ATCC 25923(3.8×10~8 cfu/m L). Effects of the extract on expressions of kat A(encoding catalase), sod A and sod M [encoding superoxide dismutases(SODs)], and ahp C [encoding alkylhydroperoxide reductase C(AhpC)] in S. aureuswere determined by RT-q PCR and corresponding enzyme activity assays were performed. Nitroblue tetrazolium reduction(NBT) assay was performed to determine effects of the extract on intracellular and extracellular levels of O_2-in S. aureus. Results: Cells challenged with 7.5 mmol/L H_2O_2 showed 0% survival in 30 min whereas 25% survived after treatment with the extract and H_2O_2. Cells that were treated with the extract alone had 43% survival in the same exposure period. Expressions of sod A and sod M genes in extract-treated cells were lowered 0.8-fold and 0.7-fold, respectively with decrease in total SOD activity of 26.8 U compared to untreated cells, 32.4 U(P<0.05). In contrast, extract-treated S. aureus cells showed 3.3-fold increase in kat A expression with corresponding increase in catalase activity of 1.828 U compared to untreated cells which was 1.248 U,(P<0.05). More profoundly, ahp C expression was increased 61-fold in extract-treated cells,(P<0.05) with corresponding increase in Ahp C activity of 0.018 U compared to untreated cells, 0.012U,(P<0.05). Extract-treated cells had significantly lower intra-and extracellular O_2-levels with absorbance readings(A_(575nm)) of 0.340 and 0.524 compared to untreated cells which were 0.516 and 0.928(P<0.05), respectively. Conclusions: Taken together these results suggest that the low MIC of A. paniculatamethanolic leaves extract(0.06 mg/m L) reduce H_2O_2 toxicity and more importantly, was in itself effectively inhibitory against S. aureus. Further, our observations suggest that a probable mode of its inhibitory mechanism against S. aureus is by reducing total SOD activity through downregulation of sod A and sod M expressions.
基金Supported by the TWAS research grant 14-062 RG/BIO/AS_GNAFOSTED grant 106-NN.02–2016.19National Research Council of Thailand(NRCT)and Center of Excellence for Innovation in Chemistry(PERCH-CIC)
文摘Objective: To isolate a-mangostin(AMG) from the peels of mangosteen(Garcinia mangostana L.), grown in Vietnam, and to investigate antibiofilm activity of this compound against three Staphylococcus aureus(S. aureus) strains, one of which was methicillin-resistant S. aureus(MRSA) and the other two strains were methicillinsensitive S. aureus(MSSA).Methods: AMG in n-hexane fraction was isolated on a silica gel column and chemically analyzed by HPLC and NMR. The antibiofilm activity of this compound was investigated by using a 96-well plate model for the formation of biofilms. Biofilm biomass was quantified using crystal violet. The viability of cells was observed under confocal microscopy using LIVE/DEAD Bac Light stains. Biofilm composition was determined using specific chemical and enzyme tests for polysaccharide, protein and DNA. Membranedamaging activity was assayed by measuring the hemolysis of human red blood cells in presence of AMG.Results: The results indicated that the isolated AMG, with a purity that exceeded 98%,had minimal inhibitory concentrations in the range of 4.6–9.2 mmol/L for the three strains tested. Interestingly, the MSSA strains were more sensitive to AMG than the MRSA strain. Minimal bactericidal concentrations were 2-fold higher than the minimal inhibitory concentration values for the three strains, indicating that AMG was a bactericidal compound. AMG also prevented biofilm formation effectively, albeit that again the MRSA strain was the most resistant. Interestingly, biofilms of the MRSA strain contained protein as a main component of the extracellular matrix, whereas this was polysaccharide in the MSSA strains. This might relate to the resistance of the MRSA 252 strain to AMG.Assays using human red blood cells indicated that AMG caused significant membrane damage with 50% of cell lysis occurred at concentration of about 36 mmol/L.Conclusions: Our results provide evidence that the isolated AMG has inhibitory activity against biofilm formation by S. aureus, including MRSA. Thus, isolated AMG proposes a high potential to develop a novel phytopharmaceutical for the treatment of MRSA.
基金the National Major R&D Program of China(grant 2016YFD0400301)the National Natural Science Foundation of China(grants 31401608).
文摘Ultrasound,is thought to a potential non-thermal sterilization technology in food industry.However,the exact mechanisms underlying microbial inactivation by ultrasound still remain obscure.In this study,the action modes of ultrasound on both Gram-negative and Gram-positive microorganisms were estimated.From colony results,ultrasound acted as an irreversible effect on both Eshcerichia coli and Staphylococcus aureus without sublethal injury.The result in this study also showed that a proportion of bacteria subpopulation suffered from serious damage of intracellular components(e.g.DNA and enzymes)but with intact cell envelopes.We speculated that the inactivated effects of ultrasound on microbes might more than simply completed disruption of cell exteriors.Those microbial cells who had not enter the valid area of ultrasonic cavitation might be injected with free radicals produced by ultrasound and experienced interior injury with intact exterior structure,and others who were in close proximity to the ultrasonic wave field would be immediately and completely disrupted into debris by high power mechanic forces.These findings here try to provide extension for the inactivation mechanisms of ultrasound on microorganisms.
基金supported by the National Natural Science Foundation of China(31972733)。
文摘Cefquinome is the only fourth-generation cephalosporin used solely for veterinary applications.In this study,we established the wild-type cut-off(CO_(WT))and pharmacokinetic/pharmacodynamic cut-off(CO_(PD))of cefquinome against Escherichia coli and Staphylococcus aureus.A total of 210 E.coli and 160 S.aureus isolates were collected from pigs in Guangdong Province between 2014 and 2018.The minimum inhibitory concentrations(MICs)were determined using a microdilution broth method.MIC_(50)and MIC_(90)were 0.06 and 0.25μg m L^(–1)for E.coli and 0.5 and 1μg m L^(–1)for S.aureus,respectively.Statistical analysis and the ECOFFinder Program showed that the CO_(WT)for cefquinome against E.coli and S.aureus were0.125 and 2μg m L^(–1),respectively.The resistance rates were 11.9%for E.coli and 6.25%for S.aureus.Based on a5000-subject Monte Carlo simulation,the CO_(PD)value for cefquinome against E.coil and S.aureus was 0.25μg m L^(–1)under the recommended dose(2 mg kg^(–1),twice a day for 3 days),confirming that infections caused by strains with MIC≤0.25μg m L^(–1)could be effectively treated.Following adjustment of the dosing regimen to 4.5 mg kg^(–1),effective treatment(>90)was achieved for S.aureus infections with MIC_(90)1μg m L^(–1).This susceptibility breakpoint determination is significant for resistant surveillance and cefquinome dosage guidance against E.coli and S.aureus in pigs.
基金financially supported by the NSFC-PSF Joint Project(31961143009)Beijing Natural Science Foundation (6182021)+2 种基金Beijing Dairy Industry Innovation Team (BAIC06)Modern Agro-industry Technology Research System (CARS-36)the Program for Changjiang Scholar and Innovation Research Team in University (IRT-15R62)。
文摘Background: Mastitis in dairy cows caused by Staphylococcus aureus is a major problem hindering economic growth in dairy farms worldwide. It is difficult to prevent or eliminate due to its asymptomatic nature and long persistence of infection. Although transcriptomic responses of bovine mammary gland cells to pathogens that cause mastitis have been studied, the common responses of peripheral blood leukocytes to S. aureus infection across two consecutive generations of dairy cattle have not been investigated.Methods: In the current study, RNA-Seq was used to profile the transcriptomes of peripheral blood leukocytes sampled from S. aureus-infected mothers and their S. aureus-infected daughters, and also healthy non-infected mothers and their healthy daughters. Differential gene expression was evaluated as follows: 1) S. aureus-infected cows versus healthy non-infected cows(S vs. H, which include all the mothers and daughters), 2) S. aureus-infected mothers versus healthy non-infected mothers(SM vs. HM), and 3) S. aureus-infected daughters versus healthy noninfected daughters(SMD vs. HMD).Results: Analysis of all identified expressed genes in the four groups(SM, SMD, HM, and HMD) showed that EPOR,IL9, IFNL3, CCL26, IL26 were exclusively expressed in both the HM and HMD groups, and that they were significantly(P < 0.05) enriched for the cytokine-cytokine receptor interaction pathway. A total of 17, 13 and 10 differentially expressed genes(DEGs)(FDR Padj. < 0.1 and |FC| > 1.2) were detected in the three comparisons, respectively. DEGs with P < 0.05 and |FC| > 2 were used for functional enrichment analyses. For the S vs. H comparison, DEGs detected included CCL20, IL13 and MMP3, which are associated with the IL-17 signaling pathway. In the SM vs. HM and SMD vs. HMD comparisons, five(BLA-DQB, C1 R, C2, FCGR1 A, and KRT10) and six(BLA-DQB, C3 AR1, CFI, FCAR, FCGR3 A, and LOC10498484) genes, respectively, were involved in the S. aureus infection pathway.Conclusions: Our study provides insights into the transcriptomic responses of bovine peripheral blood leukocytes across two generations of cattle naturally infected with S. aureus. The genes highlighted in this study could serve as expression biomarkers for mastitis and may also contain sequence variation that can be used for genetic improvement of dairy cattle for resilience to mastitis.
基金financially supported by the NSFC-PSF Joint Project(31961143009)Beijing Dairy Industry Innovation Team (BAIC06)+2 种基金China Agriculture Research System of MOF and MARABeijing Natural Science Foundation (6182021)the Program for Changjiang Scholar and Innovation Research Team in University (IRT-15R62)。
文摘Background:Staphylococcus aureus(S.aureus)mastitis is one of the most difficult diseases to treat in lactating dairy cows worldwide.S.aureus with different lineages leads to different host immune responses.Long non-coding RNAs(lncRNAs)are reported to be widely involved in the progress of inflammation.However,no research has identified stable lncRNAs among different S.aureus strain infections.In addition,folic acid(FA)can effectively reduce inflammation,and whether the inflammatory response caused by S.aureus can be reduced by FA remains to be explored.Methods:lncRNA transcripts were identified from Holstein mammary gland tissues infected with different concentrations of S.aureus(in vivo)and mammary alveolar cells(Mac-T cells,in vitro)challenged with different S.aureus strains.Differentially expressed(DE)lncRNAs were evaluated,and stable DE lncRNAs were identified in vivo and in vitro.On the basis of the gene sequence conservation and function conservation across species,key lncRNAs with the function of potentially immune regulation were retained for further analysis.The function of FA on inflammation induced by S.aureus challenge was also investigated.Then,the association analysis between these keys lncRNA transcripts and hematological parameters(HPs)was carried out.Lastly,the knockdown and overexpression of the important lncRNA were performed to validate the gene function on the regulation of cell immune response.Results:Linear regression analysis showed a significant correlation between the expression levels of lncRNA shared by mammary tissue and Mac-T cells(P<0.001,R^(2)=0.3517).lncRNAs PRANCR and TNK2-AS1 could be regarded as stable markers associated with bovine S.aureus mastitis.Several HPs could be influenced by SNPs around lncRNAs PRANCR and TNK2-AS1.The results of gene function validation showed PRANCR regulates the mRNA expression of SELPLG and ITGB2 within the S.aureus infection pathway and the Mac-T cells apoptosis.In addition,FA regulated the expression change of DE lncRNA involved in toxin metabolism and inflammation to fight against S.aureus infection.Conclusions:The remarkable association between SNPs around these two lncRNAs and partial HP indicates the potentially important role of PRANCR and TNK2-AS1 in immune regulation.Stable DE lncRNAs PRANCR and TNK2-AS1 can be regarded as potential targets for the prevention of bovine S.aureus mastitis.FA supplementation can reduce the negative effect of S.aureus challenge by regulating the expression of lncRNAs.
基金This work was supported by National Institutes of Health grants HL125352 and U01AI124316(VN).
文摘Staphylococcus aureus(S.aureus)is a leading human pathogen capable of producing severe invasive infections such as bacteremia,sepsis,and endocarditis with high morbidity and mortality,exacerbated by the increasingly widespread antibiotic resistance exemplified by methicillin-resistant strains(MRSA).S.aureus pathogenesis is fueled by the secretion of toxins—such as the membrane-damaging pore-forming atoxin,which have diverse cellular targets including the epithelium,endothelium,leukocytes,and platelets.Here,we examine the use of human platelet membrane-coated nanoparticles(PNPs)as a biomimetic decoy strategy to neutralize S.aureus toxins and preserve host cell defense functions.The PNPs blocked platelet damage induced by S.aureus secreted toxins,thereby supporting platelet activation and bactericidal activity.Likewise,the PNPs blocked macrophage damage induced by S.aureus secreted toxins,thus supporting macrophage oxidative burst,nitric oxide production,and bactericidal activity,and diminishing MRSA-induced neutrophil extracellular trap release.In a mouse model of MRSA systemic infection,PNP administration reduced bacterial counts in the blood and protected against mortality.Taken together,the results from the present work provide a proof of principle of the therapeutic benefit of PNPs in toxin neutralization,cytoprotection,and increased host resistance to invasive S.aureus infection.
