Objective: The present work was undertaken to investigate the impact of sub inhibitory concentration of gentamicin on hla gene expression in methicillin resistant Staphylococcus aureus isolates. Methods: The bacterial isolates used in this study represent 33 MRSA strains, previously isolated form patients visiting several hospitals in Baghdad. Gentamicin, vancomycin, and oxacillin MIC were determined using broth dilution method. Microtiter plate method was adopted to investigate the biofilm forming capacity. Alpha hemolysin was detected by culturing MRSA isolates on rabbit blood agar. Furthermore, hla gene was detected in MRSA isolates using conventional PCR technique; while, qRT-PCR method was performed to assay the hla expression in planktonic and biofilm cells in presence and absence of gentamicin. Results: the present results demonstrated that 12 (36.36%) isolates were gentamicin-resistant; whereas, all isolates were resistant to oxacillin and sensitive to vancomycin. Out of 33 MRSA, 3, 23, and 7 isolates formed a weak, moderate, and strong biofilm, respectively. Phenotypically, 30 isolates produced alpha hemolysin on rabbit blood agar plates; nevertheless, hla gene was located in 29 isolates. Of considerable interest, the addition of gentamicin significantly (P < 0.05) reduced the hemolysis activity; while, insignificant fold change (less than two) of hla gene was observed in all tested isolates in the presence of sub MIC of gentamicin (16 µg/ml). Conclusion: gentamicin upregulated the hla gene expression in biofilm cells; hitherto, this increment was isolate specific.
The current study aimed to detect the effect of gentamicin stress on the expression of hla (encodes hemolysin) and nuc (encodes nuclease) genes of Staphylococcus aureus. Fifty-eight isolates identified as S. aureus were isolated locally from different clinical specimens. Disk diffusion method was used to detect the resistance to S. aureus. The minimum inhibitory concentration (MIC) of gentamicin was estimated by broth microdilution method. hla and nuc genes were determined by polymerase chain reaction technique. The biofilm was evaluated using the microtiter plate method in the presence and absence of gentamicin at sub-MIC. The results showed that 18 (31%) and 40 (69%) S. aureus isolates were sensitive and resistant to gentamicin, respectiv
... Show MoreThe current study aimed to detect the effect of gentamicin stress on the expression of hla (encodes hemolysin) and nuc (encodes nuclease) genes of Staphylococcus aureus. Fifty-eight isolates identified as S. aureus were isolated locally from different clinical specimens. Disk diffusion method was used to detect the resistance to S. aureus. The minimum inhibitory concentration (MIC) of gentamicin was estimated by broth microdilution method. hla and nuc genes were determined by polymerase chain reaction technique. The biofilm was evaluated using the microtiter plate method in the presence and absence of gentamicin at sub-MIC. The results showed that 18
... Show MoreObjectives: The current work aimed to reveal the impact of gentamicin on the fibronectin binding proteins (fnbp) gene expression and its relation to biofilm and agr type in Staphylococcus aureus. Materials and Methods: A total of 25 S. aureus isolates were enrolled in this study previously isolated from different specimens. Identification confirmation and methicillin resistance were achieved by amplification of 16SrRNA and mecA. Multiplex polymerase chain reaction (PCR) based assay was employed to evaluate the agr typing. The gene expression of fnbA and fnbB genes was tested by real-time PCR technique. Minimum inhibitory concentration was estimated by micro broth dilution methodology. Microtiter plate method was performed to determine the a
... Show MoreThe pathogenicity resulting from Staphylococcus aureus infection has remarkable importance as one of the community-associated bacterial infections, due to the virulent ability of these bacteria to produce biofilms. This study was designed to detect biofilm production in clinical isolates from samples of wounds and urinary tract infections. The expression levels of the icaA gene that is responsible of slime layer production in biofilms was compared in isolates with different biofilm producing capabilities. Fifty seven samples that included 32 samples from urine and 25 samples from wounds were collected from Alwasti Hospital, Al-Kindi Teaching Hospital, and Alzahraa Clinic, Baghdad, Iraq. The bacteria was identified accor
... Show MoreNormally, bacteria exposed to antibiotics at sub minimal inhibitory concentrations (MIC) inside the host. Therefore, the current study aimed to comprehend the association among hemolysins, biofilm, as well as gentamicin resistance in local MRSA isolates. Around 35 Staphylococcus aureus locally isolated from different clinical specimens were employed in this study. Methicillin resistance was detected via cefoxitin disk diffusion and mecA amplification methods. MIC of gentamicin was estimated by broth microdilution method. Hemolysin genes involving hla, hlb, hld, and hlg were determined using multiplex polymerase chain reaction (PCR) technique. Microtiter plate method was employed for biofilm assessment in the presence and absence of gentamic
... Show MoreNormally, bacteria exposed to antibiotics at sub minimal inhibitory concentrations (MIC) inside the host. Therefore, the current study aimed to comprehend the association among hemolysins, biofilm, as well as gentamicin resistance in local MRSA isolates. Around 35 Staphylococcus aureus locally isolated from different clinical specimens were employed in this study. Methicillin resistance was detected via cefoxitin disk diffusion and mecA amplification methods. MIC of gentamicin was estimated by broth microdilution method. Hemolysin genes involving hla, hlb, hld, and hlg were determined using multiplex polymerase chain reaction (PCR) technique. Microtiter plate method was employed for biofilm assessment
... Show MoreEight isolates of methicillin resistance Staphylococcus aureus(MRSA) (SA40,SA32,SA30,SA13,SA10,SA36,SA3 and SA7) with different resistance phenotypes to macrolides , lincosamides and streptogramins Were used to detect theexpression of msrA, msrB, and linA/linA’genesby using real time polymerase chain reaction before and after treatment with antibiotics (erythromycin , clarithromycin , clindamycin and lincomycin) calibrated with triosphosphateisomerase.There highst expression of these genes was after 18 hours. It was an induction in the expression of msrA gene in isolates (SA40,SA32,SA30 and SA13) in presence of erythromycin,however,the isolates showed reduction in expression l
... Show MoreThis study included collection of 100 specimens from patients in AL-Kindy Teaching Hospital and teaching laboratories of Medical City Hospitals in Baghdad during the period from August to December 2012 ,these specimens differed in their sources which included 19 nasal swab, 16 wound swab,27 burn swab, 7 pus, 15 sputum, 10 corneal swab and 6 urine . Only 38 (38%) isolates was identified as Staphylococcus. In this study, 29 isolates (76.3%) were coagulase-positive (COPS), while only 9 isolates(23.6%) were coagulase negative (CONS), from total 38 isolates of Staphylococci.
The distribution of Methicillin resistance among Staphylococcus spp. was investigated by disc diffusion method. In this study, 21 isolates (55.26%) showed resistant to
Biofilms formation by pathogens microbial Control considered important in medical research because it is the hazarded virulence factor leading to becoming difficult to treat because of its high resistance to antimicrobials. Glycopeptide antibiotic a (Vancomycin) and the commercial bacteriocin (Nisin A) were used to comparative with purification bacteriocin (MRSAcin) against MRSA biofilm. One hundred food samples were collected from Baghdad markets from July 2016 to September 2016, including (cheese, yogurt, raw milk, fried meat, grilled meat, and beef burger). All samples were cultures; S. aureus was confirmation by macroscopic culture and microscopic examination, in addition to biochemical tests. Methicillin resistance S. asureus (
... Show MoreMethicillin resistant Staphylococcus aureus (MRSA) is the most common pathogenic bacteria in the hospitals and communities, the ability to form biofilm is considered the main cause of Staphylococcus pathogenicity since it provides resistance to both antibiotics and host immune response, so this study was aimed to evaluate the biofilms formation and its association with antibiotic resistance in clinical isolates of MRSA, in order to achieve this aim, 237 samples were collected from different patients with wounds infections after surgeries and samples from operations galleries from varies hospitals in Baghdad ,68 isolates out of 237 were subjected to Staphylococcus aureus according to conventional meth
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