Inducible Resistance Phenotype Research Articles

Unveiling the genetic landscape of Streptococcus agalactiae bacteremia: emergence of hypervirulent CC1 strains and new CC283 strains in Tehran, Iran

BackgroundThe emergence of Streptococcus agalactiae infections in patients with bacteremia is increasing. It is crucial to investigate the epidemiology, molecular characteristics, biofilm status, and virulence analysis of Streptococcus agalactiae in these patients.MethodsIn this cross-sectional study, 61 S. agalactiae isolated from blood infection were subjected to characterization through antimicrobial susceptibility tests, biofilm formation, multilocus sequence typing (MLST), and PCR analysis for detecting resistance (tet and erm family) and virulence genes (alp2/3, alp4, bca, bac, eps, rib, lmb, cylE, and pilus island genes).ResultsOverall, 32.7% of the isolates demonstrated an inducible clindamycin resistance phenotype. The results showed that 49.2, 24.6, and 8.2% of confirmed Streptococcus agalactiae strains were classified as strong, intermediate, and weak biofilm-forming strains, respectively. tet(M) (57.1%) was recovered most, followed by tet(M) + tet(L) (14.3%), tet(S) + tet(K) (10.7%), tet(M) + tet(K) (8.9%), tet(M) + tet(K) + tet(O) (5.4%), and tet(S) + tet(L) + tet(O) (3.6%). Three virulence gene profiles of cylE, lmb, bca, rib (24.6%; 15/61), cylE, lmb, rib, alp3 (19.7%; 12/61), and cylE, lmb, bac, rib (16.4%; 10/61) were detected in approximately two-thirds of the isolates. MLST revealed that the 61 isolates belonged to six clonal complexes, including CC1 (49.2%), followed by CC12 (18%), CC19 (13.1%), CC22 (9.8%), CC17 (6.6%), and CC283 (3.3%), and 11 different sequence types (STs), including ST1 (24.6%), ST7 (14.8%), ST918 (13.1%), ST2118 (9.8%), ST19 (9.8%), ST48 (6.6%), ST1372 (4.9%), ST22 (4.9%), ST40 (4.9%), ST734 (3.3%), and ST283 (3.3%). Remarkably, all CC1 and CC12 isolates, three-fourths of CC19, and half of CC22 were confirmed as biofilm producers. Conversely, CC17 and CC28 isolates were found to be nonproducers. The occurrence of strong biofilm formation was limited to specific CCs, namely CC1 (34.4%), CC12 (8.2%), CC19 (3.3%), and CC22 (3.3%).ConclusionThe high prevalence of CC1 and CC12 clones among S. agalactiae strains reflects the emergence of these lineages as successful clones in Iran, which is a serious concern and poses a potential threat to patients.

Prevalence and genetic characteristics ofStaphylococcus aureus isolates from cellphones of medical students from Iran.

Although mobile phones as a rapid communication vehicle can lead to improved quality of healthcare, they can also facilitate the transmission of pathogens to patients. This current research focuses on genetic diversity, and genes involved in resistance and biofilm production of Staphylococcus aureus isolates from mobile phones of medical students. Antibiotic resistance profiling and polymerase chain reaction (PCR) amplification of antibiotic resistance and biofilm-related genes were investigated and statistically analyzed. Staphylococcal cassette chromosome mec (SCCmec) types were analyzed by multiplex PCR, and S. aureus protein A gene typing (spa typing) was done using PCR and sequencing. Sixty-four S. aureus isolates (16.8%) were obtained from 380 medical students' mobile phones who were working in hospitals. The findings showed that 71.9% of the isolates were MRSA and 78.1% were classified as MDR. All isolates exhibited sensitivity to vancomycin and linezolid. Overall, 7.8% of the isolates displayed an inducible clindamycin resistance phenotype, while 26.7% showed resistance to mupirocin. The results indicated that 68.8% of the isolates were biofilm producers, with 7 isolates (15.9%) classified as strong producers, 22 isolates (50%) as moderate producers, and 15 isolates (34.1%) as weak producers. The most prevalent type was CC8-MRSA III/t030 (18.7%), followed by CC8-MRSA III/t037 (12.5%), CC/ST22-MSSA/t790 (10.9%), CC1-MRSA IV-t114 (9.4%), CC1-MRSA IV-t127 (7.8%), CC8-MRSA V/t064 (7.8%), CC/ST15-MSSA-t360 (7.8%), CC30-MSSA/t021(6.3%), MRSA V-t355 (6.3%), CC8-MRSA III/t421 (4.7%), CC1-MRSA V-t267 (4.7%), and CC/ST15-MSSA-t084 (3.1%). The genetic diversity and prevalent multidrug resistance indicate that the resistance situation of S. aureus recovered from mobile phones in Tehran is severe, posing a potential threat to patients, the community, and healthcare settings.

Molecular analysis of broad-spectrum induced resistance in rice by the green leaf volatile Z-3-hexenyl acetate.

Green leaf volatiles (GLVs), volatile organic compounds released by plants upon tissue damage, are key signaling molecules in plant immunity. The ability of exogenous GLV application to trigger an induced resistance (IR) phenotype against arthropod pests has been widely reported, but its effectiveness against plant pathogens is less well understood. In this study, we combined mRNA sequencing-based transcriptomics and phytohormone measurements with multispectral imaging-based precision phenotyping to gain insights into the molecular basis of Z-3-hexenyl acetate-induced resistance (Z-3-HAC-IR) in rice. Furthermore, we evaluated the efficacy of Z-3-HAC-IR against a panel of economically significant rice pathogens: Pyricularia oryzae, Rhizoctonia solani, Xanthomonas oryzae pv. oryzae, Cochliobolus miyabeanus, and Meloidogyne graminicola. Our data revealed rapid induction of jasmonate metabolism and systemic induction of plant immune responses upon Z-3-HAC exposure, as well as a transient allocation cost due to accelerated chlorophyll degradation and nutrient remobilization. Z-3-HAC-IR proved effective against all tested pathogens except for C. miyabeanus, including against the (hemi)biotrophs M. graminicola, X. oryzae pv. oryzae, and P. oryzae. The Z-3-HAC-IR phenotype was lost in the jasmonate (JA)-deficient hebiba mutant, which confirms the causal role of JA in Z-3-HAC-IR. Together, our results show that GLV exposure in rice induces broad-spectrum, JA-mediated disease resistance with limited allocation costs, and may thus be a promising alternative crop protection approach.

The Oomycete Microbe-Associated Molecular Pattern, Arachidonic Acid, and an Ascophyllum nodosum-Derived Plant Biostimulant Induce Defense Metabolome Remodeling in Tomato.

Arachidonic acid (AA) is an oomycete-derived microbe-associated molecular pattern (MAMP) capable of eliciting robust defense responses and inducing resistance in plants. Similarly, Ascophylum nodosum extract (ANE) from the brown seaweed A. nodosum, a plant biostimulant that contains AA, can also prime plants for defense against pathogen challenges. A previous parallel study comparing the transcriptomes of AA- and ANE-root-treated tomatoes demonstrated significant overlap in transcriptional profiles, a shared induced resistance phenotype, and changes in the accumulation of various defense-related phytohormones. In this work, untargeted metabolomic analysis via liquid chromatography-mass spectrometry was conducted to investigate the local and systemic metabolome-wide remodeling events elicited by AA and ANE root treatment in tomatoes. Our study demonstrated AA and ANE's capacity to locally and systemically alter the metabolome of tomatoes with enrichment of chemical classes and accumulation of metabolites associated with defense-related secondary metabolism. AA- and ANE-root-treated plants showed enrichment of fatty acyl-glycosides and strong modulation of hydroxycinnamic acids and derivatives. Identification of specific metabolites whose accumulation was affected by AA and ANE treatment revealed shared metabolic changes related to ligno-suberin biosynthesis and the synthesis of phenolic compounds. This study highlights the extensive local and systemic metabolic changes in tomatoes induced by treatment with a fatty acid MAMP and a seaweed-derived plant biostimulant with implications for induced resistance and crop improvement.

Inhibition of Erythromycin and Erythromycin-Induced Resistance among Staphylococcus aureus Clinical Isolates.

The increasing incidence of erythromycin and erythromycin-induced resistance to clindamycin among Staphylococcus aureus (S. aureus) is a serious problem. Patients infected with inducible resistance phenotypes may fail to respond to clindamycin. This study aimed to identify the prevalence of erythromycin and erythromycin-induced resistance and assess for potential inhibitors. A total of 99 isolates were purified from various clinical sources. Phenotypic detection of macrolide-lincosamide-streptogramin B (MLSB)-resistance phenotypes was performed by D-test. MLSB-resistance genes were identified using PCR. Different compounds were tested for their effects on erythromycin and inducible clindamycin resistance by broth microdilution and checkerboard microdilution methods. The obtained data were evaluated using docking analysis. Ninety-one isolates were S. aureus. The prevalence of constitutive MLSB, inducible MLSB, and macrolide-streptogramin (MS) phenotypes was 39.6%, 14.3%, and 2.2%, respectively. Genes including ermC, ermA, ermB, msrA, msrB, lnuA, and mphC were found in 82.6%, 5.8%, 7.7%, 3.8%, 3.8%, 13.5%, and 3.8% of isolates, respectively. Erythromycin resistance was significantly reduced by doxorubicin, neomycin, and omeprazole. Quinine, ketoprofen, and fosfomycin combated and reversed erythromycin/clindamycin-induced resistance. This study highlighted the significance of managing antibiotic resistance and overcoming clindamycin treatment failure. Doxorubicin, neomycin, omeprazole, quinine, ketoprofen, and fosfomycin could be potential inhibitors of erythromycin and inducible clindamycin resistance.

Diproline-induced resistance to parasitic nematodes in the same and subsequent rice generations: Roles of iron, nitric oxide and ethylene.

Induced resistance (IR) is a plant phenotype characterized by lower susceptibility to biotic challenges upon elicitation by so-called IR stimuli. Earlier, we identified diproline (cyclo(l-Pro-l-Pro)) as IR stimulus that protects rice (Oryza sativa) against the root-knot nematode Meloidogyne graminicola (Mg). In the current study, detailed transcriptome analyses at different time points, and under uninfected and nematode-infected conditions revealed that this rice IR phenotype is correlated with induction of genes related to iron (Fe), ethylene (ET) and reactive oxygen species (ROS)/reactive nitrogen species (RNS) metabolism. An infection experiment under Fe limiting conditions confirmed that diproline-IR is only effective under optimal Fe supply. Although total root Fe levels were not affected in diproline-treated plants, phytosiderophore secretion was found to be induced by this treatment. Experiments on mutant and transgenic rice lines impaired in ET or ROS/RNS metabolism confirmed that these metabolites are involved in diproline-IR. Finally, we provide evidence for transgenerational inheritance of diproline-IR (diproline-TIR), as two successive generations of diproline-treated ancestors exhibited an IR phenotype while themselves never being exposed to diproline. Transcriptome analyses on the offspring plants revealed extensive overlap between the pathways underpinning diproline-IR and diproline-TIR. Although diproline induces significant systemic changes in global DNA methylation levels early after treatment, such changes in DNA methylation were not detected in the descendants of these plants. To our knowledge, this is the first report of TIR in rice and the first transcriptional assessment of TIR in monocots.

CHARACTERIZATION OF STAPHYLOCOCCUS AUREUS ISOLATED FROM BURN WOUND; STRONG ANTIBACTERIAL ACTIVITY OF PHAGE COCKTAIL AGAINST VANCOMYCIN INTERMEDIATE-RESISTANT STAPHYLOCOCCUS AUREUS

A universal problem is about spread of Staphylococcus aureus infections in burn patients. The present study aimed to learn about the molecular characteristics and the resistance pattern of S. aureus strains isolated from burn patients. In this cross-sectional study, we investigated 100 unique S. aureus isolated from burn patients by antimicrobial activity and biofilm formation and evaluated the effect of Complex Pyobacteriophage, a commercial bacteriophage cocktail, against the isolates mentioned above. Methicillin-resistant Staphylococcus aureus (MRSA) comprised 76%, and methicillin-susceptible Staphylococcus aureus (MSSA) comprised 24% of 100 S. aureus strains. The resistance rate among MRSA isolates was higher than compared of MSSA. Mupirocin resistance was found in 30% of isolates, with 28 (93.3%) and 2 (6.7%) strains of MRSA and MSSA, respectively, found. Vancomycin intermediate resistance in S. aureus (VISA) was 13% of MRSA strains. Two isolates were confirmed as vancomycin-resistant S. aureus (VRSA) strains and carried vanA. 31 and 62% of the total isolates showed inducible and constitutive resistance phenotypes. The rate of inducible and constitutive clindamycin resistance among MRSA strains was higher than MSSA strains. Biofilm production was detected in 66% of isolates. Strong, moderate, and weak producers accounted for 25%, 17%, and 24% of isolates. Phage analysis showed that 81% were susceptible to the phage cocktail, and only 19% were resistant to the phage cocktail. Our data indicated that VISA strains prevalence in the burn unit was mainly from S. aureus infections. Present work recommended that vancomycin treatment be closely monitored to prevent the spread of VISA and VRSA strains. Observations also highlighted the role of bacteriophage cocktails in eradicating S. aureus-related infections.

Inducible clindamycin resistance in coagulase negative staphylococci

Induced clindamycin resistance poses a major problem for the use of this drug to treat staphylococcal infections. An infectious etiology is often implicated in the presence of CoNS in clinical specimens. A total of 183 isolates of Staphylococci spp. recovered from different clinical specimens were studied. A coagulase test was done to select coagulase negative Staphylococci, and they were confirmed by conventional microbiological techniques. Using the double disc approximation test, induced clindamycin resistance was detected (D-test). The D test was used to detect the inducible clindamycin resistance phenotype in erythromycin resistant and clindamycin sensitive isolates. A micro titer plate test was used to determine biofilm development in these isolates. One hundred and twenty eight isolates from various clinical specimens were identified as proven CoNS, which were distributed across six different species’. haemolyticus, the most common species of CoNS isolated, was isolated from all types of specimens. The present study revealed the strong association of biofilm with inducible clindamycin resistance among the CoNs isolates.

New update on molecular diversity of clinical Staphylococcus aureus isolates in Iran: antimicrobial resistance, adhesion and virulence factors, biofilm formation and SCCmec typing.

Staphylococcus aureus is often considered as a potential pathogen and resistant to a wide range of antibiotics. The pathogenicity of this bacterium is due to the presence of multiple virulence factors and the ability to form biofilm. SCCmec types I, II and III are mainly attributed to HA-MRSA, while SCCmec types IV and V have usually been reported in CA-MRSA infections. In this study, we performed a cross-sectional study to determine the antimicrobial resistance, adhesion and virulence factors, biofilm formation and SCCmec typing of clinical S. aureus isolates in Iran. S. aureus isolates were identified using microbiological standard methods and antibiotic susceptibility tests were performed as described by the Clinical and Laboratory Standards Institute (CLSI) guidelines. Inducible resistance phenotype and biofilm formation were determined using D-test and tissue culture plate methods, respectively. Multiplex-PCRs were performed to detect adhesion and virulence factors, antibiotic resistance genes, biofilm formation and SCCmec typing by specific primers. Among 143 clinical samples, 67.8% were identified as MRSA. All isolates were susceptible to vancomycin. The prevalence of cMLSB, iMLSB and MS phenotypes were 61.1%, 22.2% and 14.8%, respectively. The TCP method revealed that 71.3% of isolates were able to form biofilm. The predominant virulence and inducible resistance genes in both MRSA and MSSA isolates were related to sea and ermC respectively. SCCmec type III was the predominant type. Data show the high prevalence rates of virulence elements among S. aureus isolates, especially MRSA strains. This result might be attributed to antibiotic pressure, facilitating clonal selection.

Distribution of erm genes among MRSA isolates with resistance to clindamycin in a Chinese teaching hospital.

The objective of this study was to analyze erythromycin and clindamycin resistance patterns among different MRSA lineages in China. Antimicrobial susceptibility testing, resistance determinant screening, plasmid electroporation and sequence comparisons were performed. High rates of clindamycin (92.5%, 270/292) and erythromycin (92.8%, 271/292) resistance were observed. Additionally, 88.2% (60/68) of the ST59 MRSA isolates and 78.9% (15/19) of the ST239 MRSA isolates had constitutive resistance to clindamycin, while 82.0% (123/150) of the ST5 MRSA isolates showed inducible clindamycin resistance. The ermB gene was identified in 80.9% (55/68) of the ST59 isolates but was not detected in ST5 and ST239 MRSA isolates. Detection rates of ermA were high in the ST5 (99.3%, 149/150) and ST239 (89.5%, 17/19) MRSA isolates, but no ermA-positive ST59 MRSA isolates were identified. The ermC gene, observed to be harbored on similar, transmissible plasmids ranging in size from 2402 to 2473bp, were found in different MRSA lineages. Summarily, high erythromycin and clindamycin resistance rates were observed in MRSA isolates. ST59 and ST239 MRSA isolates primarily exhibited constitutive resistance, while ST5 MRSA isolates showed inducible resistance phenotypes. ermA and ermB genes were frequently carried by specific MRSA clones, while ermC gene was present within small transmissible plasmids in all lineages. Erythromycin and clindamycin resistance genes transfer between MRSA isolates in healthcare settings remains a problem, and infection control procedures should be applied.

Determination of nasal carriage and skin colonization, antimicrobial susceptibility and genetic relatedness of Staphylococcus aureus isolated from patients with atopic dermatitis in Szczecin, Poland

BackgroundAtopic dermatitis (AD) is one of the most frequent chronic and inflammatory skin condition. AD is characterized by damaged epidermal barrier, xerosis and pruritus of eczematous skin lesions which tend to flare. The duration and frequency of exacerbation of AD symptoms markedly affects the quality of patient life. AD results from the interplay between host genetics, immunity, and environmental factors, however the detailed pathogenesis of this disease is still not entirely cleared. Furthermore, disturbances of the skin microbiota and skin functional impairment predispose to secondary skin infections. Staphylococcus aureus colonizes skin and mucous membranes of 20 to 80% of healthy individuals and of 90% of patients with AD in whom this bacterium is accounted as an important AD exacerbating factor. It is also proven, that S. aureus nasal carriage significantly increases the risk for self-transmission and endogenous infection. In the current study the presence of S. aureus either in nasal vestibule and on lesioned skin of 64 patients with AD enrolled in 10-year autovaccination program was determined. The genetic relatedness of 86 S. aureus isolated from patients nose and skin using Pulsed Field Gel Electrophoresis (PFGE) and antimicrobial susceptibility of all strains to methicillin, erythromycin, clindamycin, mupirocin, gentamicin, amikacin, tetracycline, chloramphenicol and cotrimoxazole was also evaluated.ResultsIn total 23 PFGE genotypes and 24 unique patterns were distinguished. 34 patients were S. aureus nasal carriers. Simultaneous presence of S. aureus in nose and on affected skin was found in 16 carriers colonized by indistinguishable or potentially related S. aureus vs 2 carriers colonized with non-related S. aureus in nasal vestibule and on skin.4 isolates were methicillin resistant (MRSA) among which 3 showed constitutive MLSB resistance phenotype and remaining one was resistant to tetracycline and chloramphenicol.In 4 isolates inducible MLSB resistance phenotype was found, one of them was additionally resistant to tetracycline. 7 S. aureus were mupirocin resistant among them 3 - isolated from one patient, were resistant simultaneously to tetracyclines and chloramphenicol. 7 strains demonstrated resistance to chloramphenicol and susceptibility to all tested antimicrobial agents. The susceptibility to gentamicin, amikacin and cotrimoxazole among all examined S. aureus was confirmed.ConclusionThe obtained results indicated non-clonal structure of S. aureus circulating in AD patients. PFGE results showed the clonal-structure of vast majority of S. aureus isolated from nose and skin from nasal carriers what may prove the autoinfection in these patients. All examined patients the moderate or strong severity of AD was reported. Susceptibility to most antibiotics among isolated strains was also observed.

Inducible clindamycin resistance in clinical isolates of staphylococcus aureus in Suez Canal University Hospital, Ismailia, Egypt.

The increasing incidence of methicillin resistance among Staphylococci has led to renewed interest in the usage of macrolide-lincosamide-streptogramin B (MLSB) antibiotics to treat S. aureus infections, with clindamycin being the preferable agent owing to its excellent pharmacokinetic properties. Inducible clindamycin resistance my lead to therapeutic failure. Detection of the prevalence of constitutive and inducible clindamycin resistance in clinical isolates of S. aureus to improve the clinical outcomes in patients. A total of 176 non-duplicate staphylococcal isolates were isolated from different clinical samples. Methicillin resistance was detected using Cefoxitin disk diffusion (CDD) method. Phenotypic clindamycin resistance was performed for all isolates by D test. Polymerase Chain Reaction (PCR) assay were done for detection of erm resistance genes (ermA, ermB and ermC). Out of 176 strains of S. aureus, 108 isolates (61.3%) were identified as MRSA. Erythromycin and clindamycin resistance was detected in 96 isolates (54.5%) and 68 isolates (38.6%) respectively. Clindamycin resistance (cMLSB) was significantly higher (p value < 0.001) in MRSA strains (56 isolates) compared to MSSA (12 isolates). Resistant genes were detected in 160 isolates (91%). The ermA gene was detected in 28 isolates (16%), the ermB gene was detected in 80 isolates (45.5%) (p < 0.001). The frequency of constitutive and inducible clindamycin resistance in MRSA isolates emphasizes the need to use D test in routine antimicrobial susceptibility testing to detect the susceptibility to clindamycin as the inducible resistance phenotype can inhibit the action of clindamycin and affect the treatment efficacy.

Prevalence of inducible clindamycin resistance among Staphylococcus aureus isolates from a tertiary care hospital

Clindamycin is an excellent drug for skin and soft tissue Staphylococcus aureus infections, but resistance mediated by inducible macrolide-lincosamide-streptogramin B (iMLS)phenotype leads to in vivo therapeutic failure even though they may be in vitro susceptible in Kirby–Bauer disk diffusion method (KBDDM). Hence the study was undertaken to detect the prevalence of iMLS phenotype among Staphylococcus aureus isolates by double disk approximation test (D-test) in a tertiary care hospital. A total of 100 consecutive Staphylococcus species isolates were identified by standard microbiological methods and subjected to antimicrobial susceptibility testing by KBDDM. Clindamycin-resistance either in the form of iMLSB or cMLSB was determined through double disk diffusion method or D-test by using erythromycin (2 μg) and clindamycin (15μg) as per the CLSI guidelines. Out of 100 Staphylococcus species studied, 50(50%) were methicillin sensitive Staphylococcus aureus, 30(30%) were Methicillin resistant Staphylococcus aureus and 20 (20%) were Coagulase negative Staphylococci. Out of 80 Staphylococcus aureus studied, iMLSB, cMLSB and MS phenotype were 32.5%, 1.25%, 5% respectively. Inducible resistance and MS phenotype were found to be higher in MRSA as compared to MSSA (60%, 6.66% and 16%, 4% respectively). The study revealed 32.5% of Staphylococcus aureus isolates were inducible clindamycin resistant, which could be easily misidentified as clindamycin susceptible in Kirby–Bauer disk diffusion method. Therefore, clinical microbiology laboratory should routinely perform D-test in all clinically isolated Staphylococcus aureus to guide clinicians for the appropriate use of clindamycin.

Phenotypic Detection of Macrolide-Lincosamide-Streptogramin Resistance among Staphylococcus aureus and Staphylococcus epidermidis in Baghdad, Iraq

Introduction: Staphylococci emerged as the most frequent nosocomial and community-acquired pathogens. Macrolide-lincosamide-streptogramin (MLS) antibiotics resistance was increasing among Staphylococcus aureus and Staphylococcus epidermidis (S. epidermidis) isolates. Objective: In the present study, the aim was to detect the phenotypic resistance pattern of MLS (constitutive and inducible) among S. aureus and S. epidermidis isolated from Iraqi patients. Methods: A total of 120 staphylococcal isolates (60 S. aureus and 60 S. epidermidis) were isolated from urine, wound swab, blood, and sputum specimens, then specified by the VITEK 2 system. Whole isolates were investigated by the disk-diffusion method against many antibiotics, then they were checked for the MLS phenotype by the D-zone test. Results: Out of 60 S. aureus isolates and 60 S. epidermidis, the isolation rates from wound, urine, blood, and sputum were 66.6 and 50%, 16.7 and 26.7%, 11.7 and 18.3%, and 5% for each species, respectively. The higher frequency rates of resistance were showed against erythromycin, clindamycin, and streptomycin, for both S. aureus with 83.3, 53.3, and 83.3%, respectively, and S. epidermidis with 73.3, 45, and 76.7%, respectively. Constitutive MLS resistance phenotype (MLSc) was shown in 32 isolates (53.3%) of S. aureus and inducible MLS resistance phenotype (MLSi) was noted in 16 isolates (26.7%). Conclusion: The current study concluded that the D-zone test must be applied within the routine work of the antimicrobial susceptibility test for staphylococcal isolates, to exclude the false results of staphylococcal isolates sensitivity against clindamycin.

Molecular basis of macrolide-lincosamide-streptogramin (MLS) resistance in Finegoldia magna clinical isolates

Of 69 clinical isolates of Finegoldia magna tested, 36% presented high-level MICs of erythromycin (>256 μg/ml), harboring erm(A) (n = 20) or erm(B) (n=5). Of nine isolates exhibiting an inducible resistance phenotype to macrolides-lincosamides-streptogramins B, four (44%) were susceptible with a potential risk of treatment failure due to emergence of resistant mutants.

Dissecting erm(41)-Mediated Macrolide-Inducible Resistance in Mycobacterium abscessus.

Macrolides are the cornerstone of Mycobacterium abscessus multidrug therapy, despite that most patients respond poorly to this class of antibiotics due to the inducible resistance phenotype that occurs during drug treatment. This mechanism is driven by the macrolide-inducible ribosomal methylase encoded by erm(41), whose expression is activated by the transcriptional regulator WhiB7. However, it has been debated whether clarithromycin and azithromycin differ in the extent to which they induce erm(41)-mediated macrolide resistance. Herein, we show that macrolide resistance is induced more rapidly in various M. abscessus isolates upon exposure to azithromycin than to clarithromycin, based on MIC determination. Macrolide-induced expression of erm(41) was assessed in vivo using a strain carrying tdTomato placed under the control of the erm(41) promoter. Visualization of fluorescent bacilli in infected zebrafish demonstrates that azithromycin and clarithromycin activate erm(41) expression in vivo That azithromycin induces a more rapid expression of erm(41) was confirmed by measuring the β-galactosidase activity of a reporter strain in which lacZ was placed under the control of the erm(41) promoter. Shortening the promoter region in the lacZ reporter plasmid identified DNA elements involved in the regulation of erm(41) expression, particularly an AT-rich motif sharing partial conservation with the WhiB7-binding site. Mutation of this motif abrogated the macrolide-induced and WhiB7-dependent expression of erm(41). This study provides new mechanistic information on the adaptive response to macrolide treatment in M. abscessus.

Characteristics of Erythromycin Resistance in Methicillin-Resistant Staphylococcus aureus Isolated From Raw Milk

Background: Methicillin-resistant Staphylococcus aureus (MRSA) strains are one of the most important multidrug resistant microorganisms that threaten human health. Objective: The present study was conducted to evaluate genotypic and phenotypic characteristics of erythromycin resistance among MRSA isolates recovered from raw milk in Iran. Materials and Methods: A total of 50 MRSA isolates were recovered from raw milk. Tests for erythromycin and clindamycin susceptibility and inducible clindamycin resistance were done. In addition, the presence of the methicillin resistance determinant (mecA), erythromycin resistance genes (ermA, ermB, ermC and msrA) and an important virulence gene (Panton– Valentine leukocidin) were investigated using polymerase chain reaction (PCR) method. Results: Forty-eight percent (24/50) and 46% (23/50) of the isolates were resistant to erythromycin and clindamycin, respectively. Seven (14%) isolates showed inducible clindamycin resistance phenotype. The mecA gene was detected in 88% (44/50) of MRSA isolates. The incidence of the ermA, ermB, ermC and msrA genes was 14%, 64%, 12%, and 26%, respectively and the PVL gene was present in 18% (9/50) of MRSA isolates. Conclusion: According to the results of the study, the incidence of erythromycin resistance genes and inducible clindamycin-resistant MRSA strains was high in raw milk samples in Iran.

Phenotypic and genotypic identification of &lt;i&gt;Staphylococcus aureus&lt;/i&gt; resistant to clindamycin in Mansoura University Children Hospital, Egypt

Background : Clindamycin has been a good alternative drug to penicillins in the treatment of infections caused by Staphylococcus aureus but resistance to this agent has led to therapeutic failure. Inducible clindamycin resistance in staphylococci carrying erm genes may not be detectable by routine disk diffusion test. The objective of this study is to phenotypically detect clindamycin resistance in clinical S. aureus isolates and determine the prevalence of ermA, ermB and ermC carriage among these isolates. Methodology : A total of 230 non-duplicate S. aureus were isolated from children admitted to Mansoura University Children Hospital during the period January 2016 and June 2017 by conventional microbiology method. Invitro antibiotic susceptibility to selected antibiotics including erythromycin was performed by the disk diffusion technique. The „D-zone‟ test was used to phenotypically detect inducible clindamycin resistance. The presence of ermA, ermB and ermC genes was confirmed by multiplex polymerase chain reaction (m-PCR) assay. Results : One hundred and seven (46.6%) isolates were phenotypically resistant to erythromycin while 109 (47.3%) were methicillin (cefoxitin) resistant S. aureus (MRSA). The macrolidelincosamin-streptograminB (MLSB) phenotypes among the erythromycin resistant isolates were 47 (44%) inducible MLSB and 46 (43%) constitutive MLSB, while 14 (13.0%) were MS phenotype. Although, the MLSB phenotype was more predominant in MRSA (n=60, 56.1%) than MSSA (n=33, 30.7%) while the MS phenotype was more predominant in MSSA (n=9, 8.4%) than MRSA (n=5, 4.6%) isolates, the difference was not statistically significant (p=0.0777). The ermA (29.0%, n=31) and ermC (18.7%, n=20) were the most prevalent genes carried by the isolates while ermB was carried by a few (4.7%, n=5). Forty six (43%) isolates did not carry any detectable erm gene. Conclusion : In this study, both inducible and constitutive clindamycin resistance phenotypes were common among S. aureus isolates. Although the genetic basis for this may be attributed to carriage of ermA, ermB and ermC genes, a number of the resistant isolates did not carry any of these genes. Keywords : phenotypic, genotypic, macrolide-lincosamide-streptogramin B, S. aureus, children,

A Study of Inducible Clindamycin Resistance among Staphylococcus aureus Skin and Soft Tissue Infections in A Tertiary Care Hospital

ABSTRACT Objectives: Drug resistant phenotypes like MRSA are difficult to treat requiring higher group of antibiotics. Topical agents like clindamycin can be used for the therapy of MRSA. The knowledge of prevalence of inducible clindamycin resistance phenotype is essential to prevent treatment failure. Methods : A total of 204 staphylococcal isolates obtained from skin and soft tissue infections and MRSA was detected by Cefoxitin disc diffusion method and detection of Mec A gene by Polymerase chain reaction (PCR). Antibiotic susceptibility testing was performed by Kirby Baeur disc diffusion method. The Erythromycin resistant isolates were tested for D test. The differences in antibiotic susceptibility pattern between MRSA and MSSA was compared by Chi Square test using Graph pad Quick Calcs software and p value less than 0.05 was considered as significant Results : Out of the 204 Staphylococcus aureus isolates, 48 (23.5%) were identified as MRSA by Cefoxitin disc diffusion method. All these 48(23.5%) of isolates were also positive for Mec A gene by PCR. Inducible clindamycin resistance (iMLSB resistance phenotype) was observed among 24(11.7%) of the isolates. MRSA showed comparatively lesser susceptibility than MSSA (p ≤0.05). Among the MRSA inducible clindamycin resistance was seen among 11 (22.9%). Conclusions : Emergence of drug resistance warrants antibiotic susceptibility testing for all the isolates in the laboratory. Cefoxitin disc diffusion method can be used in resource constraint laboratory where PCR facilities are not available. Inducible clindamycin resistance phenotype (iMLS phenotype) must be checked for all isolates showing erythromycin resistance to prevent treatment failure. J Microbiol Infect Dis 2019; 9(3):125-128.

Resistance-related determinants in clinically relevant Staphylococcus aureus isolated from teaching therapeutic centers, Tehran, Iran

Objectives: The purpose of this study was to determine the antimicrobial resistance-related genes in Staphylococcus aureus isolated from patients referred to 14 teaching hospitals, Tehran, Iran. Methods: A total of 225 S. aureus were obtained from clinical samples in a period of 12 months. Antimicrobial resistance, minimum inhibitory concentration minimal inhibitory concentration and Inducible resistance phenotypes were determined based on the Clinical laboratory standard institute (CLSI) guidelines. PCR was performed for amplification of mecA/B/C, iles-2, ermA/B/C and ereA/B genes. Results: 39.5 and 40.8% of isolates were resistant to oxacillin and cefoxitin (FOX), respectively. The frequency of constitutive macrolide-lincosamide-streptograminB, inducible macrolide-lincosamide-streptograminB and MS phenotypes were 55.3, 28.6 and 16.1%, respectively. 40.8, 4, 7.5, 6.2, 16 and 3.1% of strains harbored the mecA, ileS-2, ermA, ermB, ermC and ereA genes, respectively. Conclusion: The frequency of methicillin-resistant S. aureus isolates in our hospitals was high and disk diffusion testing using FOX or oxacillin and/or FOX minimal inhibitory concentration E test as an alternative to PCR for identification of methicillin-resistant S. aureus is suggested. This study highlights the hypothesis that rapid testing plays an important role in antibiotic stewardship by getting patients on targeted therapy faster.