ANALYSIS OF THE ANTIBIOGRAM PROFILES OF BIOFILM FORMING STAPHYLOCOCCUS AUREUS AND ESCHERICHIA COLI

Background and Objectives: Bacteria that attach to the surfaces and produce polymeric matrix resulting in the biofilms formation are involved in a wider range of human infections. Biofilms forming Staphylococcus aureus and Escherichia coli are considered to be highly antibiotics resistant. This study aimed to analysis the antibiogram profile of biofilm forming S. aureus and E. coli in Mukalla city, Hadhramaut,Yemen. Methods: Sixty clinical isolates of S. aureus and E. coli were isolated from different clinical samples, and identified by standard bacteriological methods, then subjected to biofilm formation detection by tissue culture plate (TCP) method. The antibiotics susceptibility test was performed by disc diffusion (Kirby-Bauer) method. Chi-square test was used to analyze the data and p value< 0.05 was taken as significant. Results: Among the total isolates S. aureus and E. coli, TCP method detected 55% of isolates as strong, 25% as moderate and 25% as weak/non-biofilm producers. Biofilm forming of S. aureus developed significantly higher degrees of antibiotic resistance of amoxicillin/clavulanic acid100%, ceftazidime 95.8%, cefotaxime62.5%, cefadroxil 45.8%, ciprofloxacin 41.7% and ceftriaxone 25% with significant statistics correlation the resistance of amoxicillin/clavulanic acid and ceftazidime and bacterial biofilm production (p-value< 0.05). The rates of antibiotics resistance biofilm E. coli were 100%, 91.7%, 75%, 70.8%, 66.7%, 62.5% and 33.3%for amoxicillin/clavulanic acid, cefadroxil, cefotaxime, ceftazidime, ceftriaxone, ciprofloxacin and co-trimoxazole respectively with statistically significant correlation of cefadroxil resistance (p-value < 0.05). Conclusion: TCP method showed that S. aureus and E. coli isolates have high degree of biofilm forming ability. A high antibiotics resistance found in biofilm producers isolates than non-biofilm producers. Peer Review History: Received: 23 October 2021; Revised: 1 December; Accepted: 17 December, Available online: 15 January 2022 Academic Editor: Dr. Nuray Arı, Ankara University, Turkiye, [email protected] UJPR follows the most transparent and toughest ‘Advanced OPEN peer review’ system. The identity of the authors and, reviewers will be known to each other. This transparent process will help to eradicate any possible malicious/purposeful interference by any person (publishing staff, reviewer, editor, author, etc) during peer review. As a result of this unique system, all reviewers will get their due recognition and respect, once their names are published in the papers. We expect that, by publishing peer review reports with published papers, will be helpful to many authors for drafting their article according to the specifications. Auhors will remove any error of their article and they will improve their article(s) according to the previous reports displayed with published article(s). The main purpose of it is ‘to improve the quality of a candidate manuscript’. Our reviewers check the ‘strength and weakness of a manuscript honestly’. There will increase in the perfection, and transparency. Received file: Reviewer's Comments: Average Peer review marks at initial stage: 5.0/10 Average Peer review marks at publication stage: 7.0/10 Reviewers: Dr. Wadhah Hassan Ali Edrees, Hajja University, Yemen, [email protected] Dr. Tamer Elhabibi, Suez Canal University, Egypt, [email protected] Dr. Rawaa Souhil Al-Kayali, Aleppo University, Syria, [email protected] Rola Jadallah, Arab American University, Palestine, [email protected] Similar Articles: PREVALENCE OF STAPHYLOCOCCUS AUREUS IN DENTAL INFECTIONS AND THE OCCURRENCE OF MRSA IN ISOLATES PREVALENCE OF METHICILLIN RESISTANT STAPHYLOCOCCUS AUREUS (MRSA) AND ANTIMICROBIAL SUSCEPTIBILITY PATTERNS AT A PRIVATE HOSPITAL IN SANA'A, YEMEN

A novel phage indirectly regulates diatom growth by infecting diatom-associated biofilm-forming bacterium

Algae and heterotrophic bacteria have close and intricate interactions, which are regulated by multiple factors in the natural environment. Phages are the major factor determining bacterial mortality. However, their impacts on the algae-associated bacteria and thus on the algae-bacteria interactions are poorly understood. Here, we obtained a diatom-associated bacterium Stappia indica SNL01 that could form biofilm and had an inhibitory effect on the growth of diatom Thalassiosira pseudonana . Meanwhile, the phage SI01 with a double-stranded circular DNA genome (44,247 bp) infecting S. indica SNL01 was isolated. Phylogenetic analysis revealed that phage SI01 represents a novel member of the Podoviridae family. The phage contained multiple lysis genes encoding for cell wall lysing muramidase and spore cortex lysing SleB, as well as depolymerase-like tail spike protein. By lysing the host bacterium and inhibiting the formation of biofilm, this phage could indirectly promote the growth of the diatom. Our results shed new insights into how phages indirectly regulate algal growth by infecting bacteria closely associated with algae or in the phycosphere. IMPORTANCE The impact of phage infection on the algae-bacteria relationship in the ocean is poorly understood. Here, a novel phage infecting the diatom-associated bacterium Stappia indica SNL01 was isolated. This bacterium could form biofilm and had a negative effect on diatom growth. We revealed that this phage contained multiple lysis genes and could inhibit the formation of bacterial biofilm, thus indirectly promoting diatom growth. This study implicates that phages are not only important regulators of bacteria but also have substantial indirect effects on algae as well as the algae-bacteria relationship.

Inactivation of Spores and Vegetative Forms of Clostridioides Difficile by Chemical Biocides: Mechanisms of Biocidal Activity, Methods of Evaluation, and Environmental Aspects

Clostridioides difficile infections (CDIs) are the most common cause of acquired diseases in hospitalized patients. Effective surface disinfection, focused on the inactivation of the spores of this pathogen, is a decisive factor in reducing the number of nosocomial cases of CDI infections. An efficient disinfection procedure is the result of both the properties of the biocidal agent used and the technology of its implementation as well as a reliable, experimental methodology for assessing the activity of the biocidal active substance based on laboratory models that adequately represent real clinical conditions. This study reviews the state of knowledge regarding the properties and biochemical basis of the action mechanisms of sporicidal substances, with emphasis on chlorine dioxide (ClO2). Among the analyzed biocides, in addition to ClO2, active chlorine, hydrogen peroxide, peracetic acid, and glutaraldehyde were characterized. Due to the relatively high sporicidal effectiveness and effective control of bacterial biofilm, as well as safety in a health and environmental context, the use of ClO2 is an attractive alternative in the control of nosocomial infections of CD etiology. In terms of the methods of assessing the biocidal effectiveness, suspension and carrier standards are discussed.

Polymer-directed inhibition of reversible to irreversible attachment prevents Pseudomonas aeruginosa biofilm formation

Non-toxic, biocompatible materials that inhibit bacterial biofilm formation on implanted medical devices and so prevent infection are urgently required. Weakly amphiphilic acrylate polymers with rigid hydrocarbon pendant groups resist bacterial biofilm formation in vitro and in vivo but the biological mechanism involved is not known. By comparing biofilm formation on polymers with the same acrylate backbone but with different pendant groups, we show that poly(ethylene glycol dicyclopentenyl ether acrylate; pEGdPEA) but not neopentyl glycol propoxylate diacrylate (pNGPDA) inhibited the transition from reversible to irreversible attachment. By using single-cell tracking algorithms and controlled flow microscopy we observed that fewer Pseudomonas aeruginosa PAO1 cells accumulated on pEGdPEA compared with pNGPDA. Bacteria reaching the pEGdPEA surface exhibited shorter residence times and greater asymmetric division with more cells departing from the surface post-cell division, characteristic of reversible attachment. Migrating cells on pEGdPEA deposited fewer exopolysaccharide trails and were unable top adhere strongly. Discrimination between the polymers required type IV pili and flagella. On pEGdPEA, the lack of accumulation of cyclic diguanylate or expression of sadB were consistent with the failure to transit from reversible to irreversible attachment. Constitutive expression of sadB increased surface adhesion sufficient to enable P. aeruginosa to form biofilms in a Mot flagellar stator dependent manner. These findings were extendable to other biofilm resistant acrylates highlighting their unique ability to inhibit reversible to irreversible attachment as a mechanism for preventing biofilm-associated infections.

Small-Molecule Compound SYG-180-2-2 to Effectively Prevent the Biofilm Formation of Methicillin-Resistant Staphylococcus aureus

The resistance of methicillin-resistant Staphylococcus aureus (MRSA) has augmented due to the abuse of antibiotics, bringing about difficulties in the treatment of infection especially with the formation of biofilm. Thus, it is essential to develop antimicrobials. Here we synthesized a novel small-molecule compound, which we termed SYG-180-2-2 (C21H16N2OSe), that had antibiofilm activity. The aim of this study was to demonstrate the antibiofilm effect of SYG-180-2-2 against clinical MRSA isolates at a subinhibitory concentration (4 μg/ml). In this study, it was showed that significant suppression in biofilm formation occurred with SYG-180-2-2 treatment, the inhibition ranged between 65.0 and 85.2%. Subsequently, confocal laser scanning microscopy and a bacterial biofilm metabolism activity assay further demonstrated that SYG-180-2-2 could suppress biofilm. Additionally, SYG-180-2-2 reduced bacterial adhesion and polysaccharide intercellular adhesin (PIA) production. It was found that the expression of icaA and other biofilm-related genes were downregulated as evaluated by RT-qPCR. At the same time, icaR and codY were upregulated when biofilms were treated with SYG-180-2-2. Based on the above results, we speculate that SYG-180-2-2 inhibits the formation of biofilm by affecting cell adhesion and the expression of genes related to PIA production. Above all, SYG-180-2-2 had no toxic effects on human normal alveolar epithelial cells BEAS-2B. Collectively, the small-molecule compound SYG-180-2-2 is a safe and effective antibacterial agent for inhibiting MRSA biofilm.

Efficiency of cold atmospheric plasma, cleaning powders and their combination for biofilm removal on two different titanium implant surfaces

Objectives Biofilm removal is the decisive factor for the control of peri-implantitis. Cold atmospheric pressure plasma (CAP) can become an effective aid due to its ability to destroy and to inactivate bacterial biofilm residues. This study evaluated the cleaning efficiency of CAP, and air-polishing with glycine (APG) or erythritol (APE) containing powders alone or in combination with CAP (APG + CAP, APE + CAP) on sandblasted/acid etched, and anodised titanium implant surface. Materials and methods On respective titanium discs, a 7-day ex vivo human biofilm was grown. Afterwards, the samples were treated with CAP, APG, APE, APG + CAP, and APE + CAP. Sterile and untreated biofilm discs were used for verification. Directly after treatment and after 5 days of incubation in medium at 37 °C, samples were prepared for examination by fluorescence microscopy. The relative biofilm fluorescence was measured for quantitative analyses. Results Air-polishing with or without CAP removed biofilms effectively. The combination of air-polishing with CAP showed the best cleaning results compared to single treatments, even on day 5. Immediately after treatment, APE + CAP showed insignificant higher cleansing efficiency than APG + CAP. Conclusions CAP supports mechanical cleansing and disinfection to remove and inactivate microbial biofilm on implant surfaces significantly. Here, the type of the powder was not important. The highest cleansing results were obtained on sandblasted/etched surfaces. Clinical relevance. Microbial residuals impede wound healing and re-osseointegration after peri-implantitis treatment. Air-polishing treatment removes biofilms very effectively, but not completely. In combination with CAP, microbial free surfaces can be achieved. The tested treatment regime offers an advantage during treatment of peri-implantitis.

The Effect of Ultraviolet Treatment on TiO2 Nanotubes: A Study of Surface Characteristics, Bacterial Adhesion, and Gingival Fibroblast Response

Titanium dioxide (TiO2) nanotubes are emerging as a provocative target for oral implant research. The aim of this study was to evaluate the effect of UV on the wettability behavior, bacterial colonization, and fibroblast proliferation rate of TiO2 nanotube surfaces prepared using different anodization voltages and aimed for use as implant abutment materials. Four different experimental materials were prepared: (1) TiO2 nanotube 10 V; (2) TiO2 nanotube 15 V; (3) TiO2 nanotube 20 V; and (4) commercial pure titanium as a control group. TiO2 nanotube arrays were prepared in an aqueous electrolyte solution of hydrofluoric acid (HF, 0.5 vol.%). Different anodization voltages were used to modify the morphology of the TiO2 nanotubes. Equilibrium contact angles were measured using the sessile drop method with a contact angle meter. The investigated surfaces (n = 3) were incubated at 37 °C in a suspension of Streptococcus mutans (S. mutans) for 30 min for bacterial adhesion and 3 days for biofilm formation. Human gingival fibroblasts were plated and cultured on the experimental substrates for up to 7 days and the cell proliferation rate was assessed using the AlamarBlue assayTM (BioSource International, Camarillo, CA, USA). The data were analyzed using one-way ANOVA followed by Tukey’s post-hoc test. Water contact angle measurements on the TiO2 after UV treatment showed an overall hydrophilic behavior regardless of the anodization voltage. The ranking of the UV-treated surfaces of experimental groups from lowest to highest for bacterial adhesion was: TiO2 nanotube 20 V < Ti and TiO2 nanotube 15 V < TiO2 nanotube 10 V (p < 0.05), and for bacterial biofilm formation was: TiO2 nanotube 20 V-TiO2 nanotube 10 V < Ti-TiO2 nanotube 15 V (p < 0.05). Fibroblast cell proliferation was lower on TiO2 nanotube surfaces throughout the incubation period and UV light treatment showed no enhancement in cellular response. UV treatment enhances the wettability behavior of TiO2 nanotube surfaces and could result in lower bacterial adhesion and biofilm formation.

Estimating the Postmortem Interval of Carcasses in the Water Using the Carrion Insect, Brain Tissue RNA, Bacterial Biofilm, and Algae

The accurate estimation of postmortem interval (PMI) is crucial in the investigation of homicide cases. Unlike carcasses on land, various biological and abiotic factors affect the decomposition of carcasses in water. In addition, the insect evidence (e.g., blow flies) that is commonly used to estimate the PMI are unavailable before the carcasses float on water. Therefore, it is difficult to estimate the PMI of a carcass in water. This study aimed to explore an effective way of estimating the PMI of a carcass in water. Carrion insects, brain tissue RNA, bacterial biofilm on the skin surface, and algae in water with PMI were studied using 45 rat carcasses in a small river. The results showed that carrion insects might not be suitable for the estimation of PMI of a carcass in water since they do not have a regular succession pattern as a carcass on land, and the flies only colonized six of the carcasses. The target genes (β-actin, GAPDH, and 18S) in the brain tissue were associated with the PMI in a time-dependent manner within 1 week after death. A polynomial regression analysis was used to assess the relationship between the gene expression profiles and PMI. The correlation coefficient R2 of each regression equation was ≥ 0.924. A third-generation sequencing analysis showed that the bacteria on the skin surface of the carcass and the algae in the water samples around the carcass had a regular succession pattern, where Cryptomonas and Placoneis incased and decreased, respectively, within first 9 days. The results of this study provide a promising way to use the brain tissue RNA, bacterial biofilm, and algae to estimate the PMI of a carcass in water.