scholarly journals Analysis host-recognition mechanism of staphylococcal kayvirus ɸSA039 reveals a novel strategy that protects Staphylococcus aureus against infection by Staphylococcus pseudintermedius Siphoviridae phages

2019 ◽  
Author(s):  
Aa Haeruman Azam ◽  
Kenji Kadoi ◽  
Kazuhiko Miyanaga ◽  
Masaru Usui ◽  
Yutaka Tamura ◽  
...  

AbstractFollowing the emergence of antibiotic resistant bacteria such as methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant Staphylococcus pseudintermedius (MRSP), phage therapy has attracted significant attention as an alternative to antibiotic treatment. Bacteriophages belonging to kayvirus (previously known as Twort-like phages) have broad host range and are strictly lytic in Staphylococcus spp. Previous work revealed that kayvirus ɸSA039 has a host-recognition mechanism distinct from those of other known kayviruses: most of kayviruses use the backbone of wall teichoic acid (WTA) as their receptor; by contrast, ɸSA039 uses the β-N-acetylglucosamine (β-GlcNAc) residue in WTA. In this study, we found that ɸSA039 could switch its receptor to be able to infect S. aureus lacking the β-GlcNAc residue by acquiring a spontaneous mutation in open reading frame (ORF) 100 and ORF102. Moreover, ɸSA039 could infect S. pseudintermedius, which has a different WTA structure than S. aureus. By comparison with newly isolated S. pseudintermedius–specific phage (SP phages), we determined that glycosylation in WTA of S. pseudintermedius is essential for adsorption of SP phages, but not ɸSA039. Finally, we describe a novel strategy of S. aureus which protects the bacteria from infection of SP phages. Notably, glycosylation of ribitol phosphate (RboP) WTA by TarM or/and TarS prevents infection of S. aureus by SP phages. These findings could help to establish a new strategy for treatment of S. aureus and S. pseudintermedius infection, as well as provide valuable insights into the biology of phage–host interactions.


2014 ◽  
Vol 5 (4) ◽  
pp. 389-395 ◽  
Author(s):  
S. Warrack ◽  
P. Panjikar ◽  
M. Duster ◽  
N. Safdar

Methicillin-resistant Staphylococcus aureus (MRSA) is a pathogen of major public health importance. Colonisation precedes infection; thus reducing MRSA carriage may be of benefit for reducing infection. Probiotics represent a novel approach to reducing MRSA carriage. We undertook a pilot feasibility randomised controlled trial of the tolerability and acceptability of probiotics for reducing nasal and intestinal carriage of MRSA. In addition, subjects were screened for vancomycin-resistant enterocococci (VRE). Subjects with a history of MRSA were recruited from a large, academic medical center and randomised to take either a placebo or probiotic (Lactobacillus rhamnosus HN001). Subjects returned to the clinic after four weeks for further testing to determine adherence to the probiotic regimen and colonisation of MRSA. 48 subjects were enrolled and randomised. Nearly 25% were transplant recipients and 30% had diabetes. The probiotic was well tolerated in the study population though minor side effects, such as nausea and bloating, were observed. A majority of the subjects randomised to HN001 had good adherence to the regimen. At the four week time point among subjects randomised to the probiotic, MRSA was detected in 67 and 50% of subjects colonised in the nares and the gastrointestinal tract, respectively. Three subjects who initially tested positive for VRE were negative after four weeks of probiotic exposure. Probiotics were well tolerated in our study population of largely immunocompromised subjects with multiple comorbidities. Adherence to the intervention was good. Probiotics should be studied further for their potential to reduce colonisation by multidrug resistant bacteria.



2011 ◽  
Vol 32 (11) ◽  
pp. 1057-1063 ◽  
Author(s):  
Nisha Nair ◽  
Ekaterina Kourbatova ◽  
Katharine Poole ◽  
Charmaine M. Huckabee ◽  
Patrick Murray ◽  
...  

Background.The multicenter, cluster-randomized Strategies to Reduce Transmission of Antimicrobial Resistant Bacteria in Intensive Care Units (STAR*ICU) trial was performed in 18 U.S. adult intensive care units (ICUs). It evaluated the effectiveness of infection control strategies to reduce the transmission of methicillin-resistant Staphylococcus aureus (MRSA) colonization and/or infection. Our study objective was to examine the molecular epidemiology of MRSA and assess the prevalence and risk factors for community acquired (CA)-MRSA genotype nasal carriage at the time of ICU admission.Methods.Selected MRSA isolates were subjected to molecular typing using pulsed-field gel electrophoresis.Results.Of 5,512 ICU patient admissions in the STAR*ICU trial during the intervention period, 626 (11%) had a nares sample culture result that was positive for MRSA. A total of 210 (34%) of 626 available isolates were selected for molecular typing by weighted random sampling. Of 210 patients, 123 (59%) were male; mean age was 63 years. Molecular typing revealed that 147 isolates (70%) were the USAIOO clone, 26 (12%) were USA300, 12 (6%) were USA500, 8 (4%) were USA800, and 17 (8%) were other MRSA genotypes. In a multivariate analysis, patients who were colonized with a CA-MRSA genotype (USA300, USA400, or USA1000) were less likely to have been hospitalized during the previous 12 months (PR [prevalence ratio], 0.39 [95% confidence interval (CI), 0.21-0.73]) and were less likely to be older (PR, 0.97 [95% CI, 0.95-0.98] per year) compared with patients who were colonized with a healthcare-associated (HA)-MRSA genotype.Conclusion.CA-MRSA genotypes have emerged as a cause of MRSA nares colonization among patients admitted to adult ICUs in the United States. During the study period (2006), the predominant site of CA-MRSA genotype acquisition appeared to be in the community.



2010 ◽  
Vol 76 (15) ◽  
pp. 5165-5174 ◽  
Author(s):  
Kanako Ishihara ◽  
Natsumi Shimokubo ◽  
Akie Sakagami ◽  
Hiroshi Ueno ◽  
Yasukazu Muramatsu ◽  
...  

ABSTRACT Recently, methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant Staphylococcus pseudintermedius (MRSP) have been increasingly isolated from veterinarians and companion animals. With a view to preventing the spread of MRSA and MRSP, we evaluated the occurrence and molecular characteristics of each in a veterinary college. MRSA and MRSP were isolated from nasal samples from veterinarians, staff members, and veterinary students affiliated with a veterinary hospital. Using stepwise logistic regression, we identified two factors associated with MRSA carriage: (i) contact with an identified animal MRSA case (odds ratio [OR], 6.9; 95% confidence interval [95% CI], 2.2 to 21.6) and (ii) being an employee (OR, 6.2; 95% CI, 2.0 to 19.4). The majority of MRSA isolates obtained from individuals affiliated with the veterinary hospital and dog patients harbored spa type t002 and a type II staphylococcal cassette chromosome mec (SCCmec), similar to the hospital-acquired MRSA isolates in Japan. MRSA isolates harboring spa type t008 and a type IV SCCmec were obtained from one veterinarian on three different sampling occasions and also from dog patients. MRSA carriers can also be a source of MRSA infection in animals. The majority of MRSP isolates (85.2%) carried hybrid SCCmec type II-III, and almost all the remaining MRSP isolates (11.1%) carried SCCmec type V. MRSA and MRSP were also isolated from environmental samples collected from the veterinary hospital (5.1% and 6.4%, respectively). The application of certain disinfection procedures is important for the prevention of nosocomial infection, and MRSA and MRSP infection control strategies should be adopted in veterinary medical practice.



2021 ◽  
Vol 12 ◽  
Author(s):  
Tingting Feng ◽  
Sebastian Leptihn ◽  
Ke Dong ◽  
Belinda Loh ◽  
Yan Zhang ◽  
...  

Phage therapy represents a possible treatment option to cure infections caused by multidrug-resistant bacteria, including methicillin and vancomycin-resistant Staphylococcus aureus, to which most antibiotics have become ineffective. In the present study, we report the isolation and complete characterization of a novel phage named JD219 exhibiting a broad host range able to infect 61 of 138 clinical strains of S. aureus tested, which included MRSA strains as well. The phage JD419 exhibits a unique morphology with an elongated capsid and a flexible tail. To evaluate the potential of JD419 to be used as a therapeutic phage, we tested the ability of the phage particles to remain infectious after treatment exceeding physiological pH or temperature. The activity was retained at pH values of 6.0–8.0 and below 50°C. As phages can contain virulence genes, JD419’s complete genome was sequenced. The 45509 bp genome is predicted to contain 65 ORFs, none of which show homology to any known virulence or antibiotic resistance genes. Genome analysis indicates that JD419 is a temperate phage, despite observing rapid replication and lysis of host strains. Following the recent advances in synthetic biology, JD419 can be modified by gene engineering to remove prophage-related genes, preventing potential lysogeny, in order to be deployed as a therapeutic phage.



2018 ◽  
Author(s):  
◽  
Johmari Logtenberg

Background: Healthcare-associated infections (HCAIs) are a global concern as they affect millions of people worldwide. Poor hygiene practices and the use of microbial contaminated medical equipment by healthcare workers (HCWs) are common contributing factors to the development of HCAIs, which result in additional hospital costs, prolonged hospital stays, development of antibacterial resistance and increases in mortality and morbidity. Because Chiropractic students (CSs) at the Durban University of Technology Chiropractic Day Clinic (DUT CDC) make use of diagnostic equipment during their consultations with patients, this study aimed to determine if the stethoscopes and sphygmomanometers that were used in the DUT CDC served as reservoirs for bacterial growth, including antibiotic-resistant bacteria, and to correlate the findings with the hygiene practices of CSs and the efficacy of disinfectants. Method: This quantitative study comprised of two phases: a phase one pre-test post-test design and a phase two cross-sectional descriptive questionnaire design. Phase one required the collection of bacterial samples from the stethoscopes and sphygmomanometers of 29 CSs before and after performing the physical assessments on new patients (58 samples). These bacterial samples were incubated and analysed. The bacterial isolates were enumerated, identified and, where appropriate, tested for antibiotic-resistance. The modified AOAC use dilution method was used to test the efficacy of the selected disinfectants. Phase two required 29 CSs to complete the research questionnaire. The data were initially captured onto Excel spreadsheets and subsequently analysed using IBM SPSS version 24.0 (p-value <0.05 was considered statistically significant), with the application of Spearman’s rank correlation, an one-way ANOVA evaluation, Tukey post hoc and paired t-tests. Results: Although the majority of the CSs was knowledgeable and regarded disinfection as important, only 13.8% applied adequate disinfection practices. The most common reasons that were stated were inadequate education or training, forgetfulness, lack of time, and disinfectant unavailability. Bacterial growth was present on 96.6% and 100.0% of the pre-test and post-test stethoscope samples respectively, and on 94.8% and 100.0% of the pre- and post-test sphygmomanometer samples respectively. The total colony-forming units (CFUs) for both the post- test readings were higher compared to their respective pre-test samples. Paired t-tests indicated significantly (p< 0.01) higher mean values for the post-stethoscope group only, with a greater distribution of the total CFUs for stethoscope samples at the diaphragms’ edge. The bacteria that were isolated from both sets of pre- and post-test samples consisted predominantly of coagulase negative staphylococci (CoNS), Micrococcus spp. and Staphylococcus aureus, while the minority consisted of Bacillus spp., Corynebacterium spp., coliforms and Escherichia coli. Overall, the majority of the bacteria was considered potentially pathogenic, except for the post-test sphygmomanometer sample. The disinfectant efficacy testing revealed results that were in contrast with the literature, which caused the researcher to question the validity and reliability of the modified AOAC use dilution method in this study. Chloramphenicol was the most effective antibiotic with a bacterial susceptibility rate of 95.7%, (Ciprofloxacin (93.2%), Vancomycin (80.8%), Amoxicillin (AMO) (69.5%) and Erythromycin (57.7%)). The resistance of Micrococcus spp. isolates to the various antibiotics was of concern. The high resistance levels of CoNS and S. aureus to AMO suggests the presence of methicillin-resistant Staphylococcus aureus and methicillin-resistant CoNS isolates. The most susceptible specie in general was Micrococcus spp. at 60.9%, followed by S. aureus at 59.6%, E. coli at 53.8%, and CoNS at 48.7%, while the least susceptible was coliforms at 36.9%. No correlations (p> 0.05) were identified between the mean CFU isolates from the CSs’ equipment and their reported average disinfection rate. Conclusions and recommendations: Both stethoscopes and sphygmomanometers were contaminated with non-pathogenic and potential pathogenic bacteria (some were resistant to multiple antibiotic classes). Although knowledgeable about equipment disinfection procedures, only 13.8% of the CSs reported disinfecting their pieces of equipment after examining every patient. The provision of adequate equipment disinfection education, the placement of visual reminders and accessible disinfectants will assist in improving the practice of adequate equipment disinfection. Moreover, equipment disinfection before and after every patient consultation will minimise cross- contamination and thus the risk of the development of HCAIs.



PLoS ONE ◽  
2021 ◽  
Vol 16 (10) ◽  
pp. e0258607
Author(s):  
Shivankar Agrawal ◽  
Jusna Nandeibam ◽  
Indira Sarangthem

Staphylococcus aureus and Methicillin-resistant S. aureus (MRSA) remains one of the major concerns of healthcare associated and community-onset infections worldwide. The number of cases of treatment failure for infections associated with resistant bacteria is on the rise, due to the decreasing efficacy of current antibiotics. Notably, Acrophialophora levis, a thermophilous fungus species, showed antibacterial activity, namely against S. aureus and clinical MRSA strains. The ethyl acetate extract of culture filtrate was found to display significant activity against S. aureus and MRSA with a minimum inhibitory concentration (MIC) of 1 μg/mL and 4 μg/mL, respectively. Scanning electron micrographs demonstrated drastic changes in the cellular architecture of metabolite treated cells of S. aureus and an MRSA clinical isolate. Cell wall disruption, membrane lysis and probable leakage of cytoplasmic are hallmarks of the antibacterial effect of fungal metabolites against MRSA. The ethyl acetate extract also showed strong antioxidant activity using two different complementary free radicals scavenging methods, DPPH and ABTS with efficiency of 55% and 47% at 1 mg/mL, respectively. The total phenolic and flavonoid content was found to be 50 mg/GAE and 20 mg/CAE, respectively. More than ten metabolites from different classes were identified: phenolic acids, phenylpropanoids, sesquiterpenes, tannins, lignans and flavonoids. In conclusion, the significant antibacterial activity renders this fungal strain as a bioresource for natural compounds an interesting alternative against resistant bacteria.



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