Staphylococcus aureus and Pseudomonas aeruginosa are the major globally distributed pathogens, which causes chronic and recalcitrant infections due to their capacity to produce biofilms in large part. Biofilm production represents a survival strategy in these species, allowing them to endure environmental stress by altering their gene expression to match their own survival needs. In this study, we co-cultured different clinical isolates of S. aureus and P. aeruginosa as mono- and mixed-species biofilms in a full-strength Brain Heart Infusion Broth (BHI) and in a 1000-fold diluted Brain Heart Infusion Broth (BHI/1000) using Microtiter plate assay and determination of colony-forming units. Furthermore, the effect of starvation stress on the expression of pslA and fnbA genes of both species was investigated using RT-PCR. This work indicated that starvation stress significantly increased the biofilm biomass and bacterial density in all mono and mixed biofilm-producing strains. Interestingly, co-culture biofilms exhibited higher resistance to starvation as compared to monoculture. The current results also showed that the expression of fnbA and pslA genes was up-regulated under starvation stress in mono-and coculture biofilm. meanwhile, up-regulates of both genes in co-culture biofilm was significantly higher than mono- species biofilm.
Bacteria form complex and highly elaborate surface adherent communities known as biofilms.Biofilm have been shown to be associated with several human diseases ,and to colonize a wide variety of medical devices . The current study focuses on contribution of extracted genomic DNA in biofilm formation by P. aeruginosa and K. pneumoniae isolates .The percentages of Pseudomonas aeruginosa recovery from drinking water in this study were 10%(20 positive P. aeruginosa samples ) and K. pneumonia., 7%(14 positive K. pneumonia samples).The results showed that all P.aeruginosa and K. pneumoniae isolates (100%) were slime producer but in different degrees by forming of black
... 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 MorePseudomonas aeruginosa, a ubiquitous environmental organism, is a difficult-to-treat opportunistic pathogen due to its broad-spectrum antibiotic resistance and its ability to form biofilms. In this study, we investigate the link between resistance to a clinically important antibiotic, imipenem, and biofilm formation. First, we observed that the laboratory strain P. aeruginosa PAO1 carrying a mutation in the oprD gene, which confers resistance to imipenem, showed a modest reduction in biofilm formation.We also observed an inverse relationship between imipenem resistance and biofilm formation for imipenem-resistant strains selected in vitro, as well as for clinical isolates.We identified two clinical isolates of P. aeruginosa from the sputum
... Show MoreThe current study was designed to explore the association between the pigments production and biofilm construction in local Pseudomonas aeruginosa isolates. Out of 143 patients suffering from burns, urinary tract infections (UTI), respiratory tract infections and cystic fibrosis obtained from previous study by Mahmood (2015), twenty two isolates (15.38%) were identified from (11) hospitals in Iraq, splitted into three provinces, Baghdad, Al-Anbar and Karbala for the duration of June 2017 to April 2018. Characterization was carried out by using microscopical, morphological and biochemical methods which showed that all these isolates belong to P. aeruginosa. Screening of biofilm production isolates was carried out by usi
... Show MorePseudomonas aeruginosa is a Gram-negative opportunistic pathogen and a model bacterium for studying virulence and bacterial social traits. While it can be isolated in low numbers from a wide variety of environments including soil and water, it can readily be found in almost any human/animal-impacted environment. It is a major cause of illness and death in humans with immunosuppressive and chronic conditions, and infections in these patients are difficult to treat due to a number of antibiotic resistance mechanisms and the organism’s propensity to form multicellular biofilms. One hundred twenty clinical samples and forty hospital environmental samples (various sources) were collected from hospitals in Baghdad city during the period from Oc
... Show MoreOne hundred isolates of Pseudomonas aeruginosa were obtained from patients admitted to Baghdad hospitals, Iraq during the period between May 2018 until July 2018. These isolates were distributed as 15 isolates from blood, 25 isolates from urinary tract infections, 10 isolates from sputum, 12 isolates from wounds, 15 isolates from ear infections, 15 isolates from bronchial wash of patients suffering from respiratory tract infections in addition to 8 isolates from cystic fibrosis patients. The isolates were initially identified by culturing on MacConkey agar, blood agar and P. aeruginosa agar then diagnosed by performing some morphological and biochemical tests. The second diagnosis was done by API 20E system followed by Vitek 2 compact syste
... Show MoreOwing to high antibacterial resistance of Pseudomonas aeruginosa, it could be considered as the main reason behind the nosocomial infections. P. aeruginosa has a well-known biofilm forming ability. The expression of polysaccharide encoding locus (pelA gene) by P. aeruginosa is essential for this ability. The purpose of the current research was to determine the biofilm formation in P. aeruginosa isolated from clinical samples and to evaluate the role of the selected PelA gene in biofilm formation using PCR method in Iraqi patients. Results revealed that 24 (96%) isolates were found to have the ability to form biofilm that was remarkably related to gentamicin resistance. Moreover, the pelA gene was found in all biofilm-producers. In c
... Show MoreObjective: 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 plank
... Show MoreOne of the most important virulence factors in Pseudomonas aeruginosa is biofilm formation, as it works as a barrier for entering antibiotics into the bacterial cell. Different environmental and nutritional conditions were used to optimize biofilm formation using microtitre plate assay by P. aeruginosa. The low nutrient level of the medium represented by tryptic soy broth (TSB) was better in biofilm formation than the high nutrient level of the medium with Luria Broth (LB). The optimized condition for biofilm production at room temperature (25 °C) is better than at host temperature (37 °C). Moreover, the staining with 0.1% crystal violet and reading the biofilm with wavelength 360 are considered essential factors in
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