Journal of Microbiology

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Anti protein A antibody-gold nanorods conjugate: a targeting agent for selective killing of methicillin resistant Staphylococcus aureus using photothermal therapy method: Rasoul Shokri , Mojtaba Salouti , Rahim Sorouri Zanjani; J. Microbiol. 2015;53(2):116-121. Published online January 28, 2015; DOI: https://doi.org/10.1007/s12275-015-4519-4

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The high prevalence of methicillin resistant Staphylococcus aureus (MRSA) and developing resistance to antibiotics requires new approaches for treatment of infectious diseases due to this bacterium. In this study, we developed a targeting agent for selective killing of MRSA using photothermal therapy method based on anti protein A antibody and gold nanorods (GNRs). Polystyrene sulfonate (PSS) coated GNRs were conjugated with anti protein A antibody. The FT-IR and UV-vis analyses approved the formation of anti protein A antibody-gold nanorods conjugate. In vitro study of photothermal therapy showed 82% reduction in the MRSA cells viability which was significantly greater than the ablation effect of free GNRs and laser alone. Significant accumulation of anti protein A antibody-GNRs in the infected muscle in comparison with normal muscle approved the targeting ability of new agent. In vivo study of photothermal therapy resulted in a significant reduction (73%) in the bacterial cells viability in the infected mouse model. These results demonstrated the ability of anti protein A antibody-GNRs conjugate in combination with NIR laser energy for selective killing of MRSA in mouse model.

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Infection Microenvironment‐Sensitive Photothermal Nanotherapeutic Platform to Inhibit Methicillin‐Resistant Staphylococcus aureus Infection
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Identification and Methicillin Resistance of Coagulase-Negative Staphylococci Isolated from Nasal Cavity of Healthy Horses: Jolanta Karakulska , Karol Fijałkowski , Paweł Nawrotek , Anna Pobucewicz , Filip Poszumski , Danuta Czernomysy-Furowicz; J. Microbiol. 2012;50(3):444-451. Published online June 30, 2012; DOI: https://doi.org/10.1007/s12275-012-1550-6

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Abstract: The aim of this study was an analysis of the staphylococcal flora of the nasal cavity of 42 healthy horses from 4 farms, along with species identification of CoNS isolates and determination of resistance to 18 antimicrobial agents, particularly phenotypic and genotypic methicillin resistance. From the 81 swabs, 87 staphylococci were isolated. All isolates possessed the gap gene but the coa gene was not detected in any of these isolates. Using PCR-RFLP of the gap gene, 82.8% of CoNS were identified: S. equorum (14.9%), S. warneri (14.9%), S. sciuri (12.6%), S. vitulinus (12.6%), S. xylosus (11.5% ), S. felis (5.7%), S. haemolyticus (3.4%), S. simulans(3.4%), S. capitis (1.1%), S. chromogenes (1.1%), and S. cohnii subsp. urealyticus (1.1%). To our knowledge, this was the first isolation of S. felis from a horse. The species identity of the remaining Staphylococcus spp. isolates (17.2%) could not be determined from the gap gene PCR-RFLP analysis and 16S rRNA gene sequencing data. Based on 16S-23S intergenic transcribed spacer PCR, 11 different ITS-PCR profiles were identified for the 87 analyzed isolates. Results of API Staph were consistent with molecular identification of 17 (19.5%) isolates. Resistance was detected to only 1 or 2 of the 18 antimicrobial agents tested in the 17.2% CoNS isolates, including 6.9% MRCoNS. The mecA gene was detected in each of the 5 (5.7%) phenotypically cefoxitin-resistant isolates and in 12 (13.8%) isolates susceptible to cefoxitin. In total, from 12 horses (28.6%), 17 (19.5%) MRCoNS were isolated. The highest percentage of MRCoNS was noted among S. sciuri isolates (100%).

Research Support, Non-U.S. Gov'ts

Responses in the Expression of Extracellular Proteins in Methicillin-Resistant Staphylococcus aureus Treated with Rhodomyrtone: Monton Visutthi , Potjanee Srimanote , Supayang Piyawan Voravuthikunchai; J. Microbiol. 2011;49(6):956-964. Published online December 28, 2011; DOI: https://doi.org/10.1007/s12275-011-1115-0

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31 Scopus

Abstract: Rhodomyrtone from a medicinal plant species, Rhodomyrtus tomentosa, is a challenged effective agent against Gram-positive bacteria, especially methicillin-resistant Staphylococcus aureus (MRSA). The present study was undertaken to provide insight into MRSA extracellular protein expression following rhodomyrtone treatment. Secreteomic approach was performed on a representative clinical MRSA isolate exposing to subinhibitory concentration rhodomyrtone (0.174 μg/ml). The identified extracellular proteins of a response of MRSA to rhodomyrtone treated condition were both suppressed and overexpressed. Staphylococcal antigenic proteins, immunodominant antigen A (IsaA) and staphylococcal secretory antigen (SsaA) involved in cell wall hydrolysis were downregulated after the treatment. The results suggested that rhodomyrtone may interfere with WalK/WalR (YycG/YycF) system. Other enzymes such as lipase precursor and another lipase, glycerophosphoryl diester phosphodiesterase, were absent. In contrast, cytoplasmic proteins such as SpoVG and glycerol phosphate lipoteichoic acid synthase, and ribosomal proteins were found in the treated sample. Appearance of several cytoplasmic proteins in the treated culture supernatant revealed that the bacterial cell wall biosynthesis was disturbed. This finding provides a proteomic mapping of extracellular proteins after rhodomytone treatment. Extensive investigation is required for this natural compound as it has a great potency as an alternative anti-MRSA drug.

Antimicrobial Susceptibilities of Coagulase-Negative Staphylococci (CNS) and Streptococci from Bovine Subclinical Mastitis Cases: Emel Banu Buyukunal Bal , Suleyman Bayar , Mehmet Ali Bal; J. Microbiol. 2010;48(3):267-274. Published online June 23, 2010; DOI: https://doi.org/10.1007/s12275-010-9373-9

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Abstract: The prevalence and antimicrobial susceptibilities of Staphylococci and Streptococci were assessed from subclinical mastitis cases. One hundred Coagulase-Negative Staphylococci (CNS) and 34 Streptoccocci were identified. The most frequently isolated species were Staphylococcus haemolyticus (27%) and Staphylococcus simulans (24%). Susceptible CNS species revealed the highest resistance to penicillin G (58%), ampicillin (48%), neomycin (20%), and oleandomycin (14%). CNS methicillin resistance rates within 82 isolates were 21.95% and 1.22% by disk diffusion and PCR methods, respectively. These results suggested the disk diffusion
method
was more prone to yield false positives. Partial sequencing of the 16S rRNA region from the mecA carrying isolate (S. haemolyticus) was homologous with S. haemolyticus sequences/accessions obtained from GenBank. However, the mecA gene sequence from this isolate was more closely allied with the S. aureus mecA gene of human origins. Identical sequence data was acquired from the National Center for Biotechnology Information (NCBI) database, suggesting horizontal gene transfer between the two species. CNS β-lactamase activity within 81 isolates was 29.63%. The most frequently isolated Streptococcus species were S. uberis (52%) and S. agalactiae (15%). Oleandomycin was the least effective antimicrobial agent on these isolates with 59% susceptibility. Results indicated that CNS and Streptococci exhibited various antimicrobial resistance responses. Consequently, isolation and identification of udder pathogens in herds suffering from subclinical agents is essential to select the most effective antimicrobial agent. Moreover, multiple resistance features of methicillin resistant (MR) isolates should be considered during antimicrobial susceptibility tests.

Synergistic Effects of the Combination of Galangin with Gentamicin against Methicillin-Resistant Staphylococcus aureus: Young-Seob Lee , Ok-Hwa Kang , Jang-Gi Choi , You-Chang Oh , Hee-Sung Chae , Jong Hak Kim , Hyun Park , Dong Hwan Sohn , Zheng-Tao Wang , Dong-Yeul Kwon; J. Microbiol. 2008;46(3):283-288. Published online July 5, 2008; DOI: https://doi.org/10.1007/s12275-008-0012-7

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Abstract: The antimicrobial killing activity toward methicillin-resistant Staphylococcus aureus (MRSA) has been a serious emerging global issue. New effective antimicrobials and/or new approaches to settle this issue are urgently needed. The oriental herb, Alpinia officinarum, has been used in Korea for several hundreds of years to treat various infectious diseases. As it is well known, one of the active constituents of Alpinia officinarum is galangin. Against the 17 strains, the minimum inhibitory concentrations (MICs) of galangin (GAL) were in the range of 62.5~125 μg/ml, and the MICs of gentamicin (GEN) ranged from 1.9 μg/ml to 2,000 μg/ml. The fractional inhibitory concentrations (FICs) of GAL, in combination with GEN, against 3 test strains were 0.4, 3.9, and 250 μg/ml, and were all 15.62 μg/ml in GEN. The FIC index showed marked synergism in the value range of 0.19 to 0.25. By determining time-kill curves, also confirmed the low synergism of the GAL and GEN combination against 4 h, 8 h, 12 h, and 24 h cultured MRSA. The time-kill study results indicated a low synergistic effect against 3 test strains. Thus, the mixture of GAL and GEN could lead to the development of new combination antibiotics against MRSA infection.

Staphylococcal methicillin resistance expression under various growth conditions: Lee, Yoo Nik , Poo Ha Ryoung , Lee, Young Ik; J. Microbiol. 1997;35(2):103-108.

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Abstract: To improve the detection of methicillin resistant staphylococci, lowered incubation temperature (30℃) and inclusion of sodium chloride in media have been empirically recommended. However, in this study, we found that sodium chloride in Peptone-Yeast Extract-K₂HPO₄(PYK) medium decreased methicillin minimum inhibitory concentrations. Divalent cations were shown to restore the expression of staphylococcal methicillin resistance. However, when it was determined by efficiency of plating, sodium chloride increased methicillin resistance expression on agar medium in which higher divalent cations were contained in the agar medium. The decrease of minimum inhibitory concentrations at 30℃ by sodium chloride occurred in Brain Heart Infusion but did not occur in other media investigated. Interestingly, both PYK and Brain Heart Infusion media had peptone, which contain cholic acids having detergent activities. Inclusion of sodium chloride in PYK caused a higher rate of autolysis. Penicillin binding protein 2a that has a low affinity to beta-lactam antibiotics, was highly inducible in methicillin resistant Staphylococcus epidermidis strains. In this study, we found that autolysins that are activated by the sodium chloride decreased the minimum inhibitory concentration at 30℃, and peptidoglycan is weakened due to the presence of methicillin. Peptone in the media may aggravate the fragile cells. However, stabilization due to the presence of divalent cations and production of penicilin binding protein 2a increase the survival of staphylococci.

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