Inhibitory Effects of Carvacrol on Biofilm Formation in Colistin Heteroresistant Acinetobacter baumannii Clinical Isolates.

BACKGROUND: The ability of bacteria to form biofilm is an essential strategy for creating stable infections. This issue is more critical in Acinetobacter bauamannii as a hospital pathogen. Today, the control of biofilm formation and solutions to prevent or remove biofilm is being developed. Carvacrol has been considered an anti-biofilm compound in significant bacteria. This study investigated the anti-biofilm effect of Carvacrol on biofilm formation in clinical colistin heteroresistant isolates of A. baumannii. METHODS: 22 clinical strains of A. baumannii were collected from Motahari Hospital in Tehran, Iran, in 2019. Biochemical and genotypic methods confirmed these isolates. Colistin heteroresistance was determined by the Standard PAP method. Carvacrol's antibacterial and anti-biofilm activity was determined according to the standard protocol. RESULTS: About 12 isolates were considered strong biofilm producers and were used for analysis. Six isolates had hetero-resistance to colistin. Carvacrol at a 512 g/ml concentration showed the best antibacterial activity against all isolates. The sub-MIC of Carvacrol (256 g/ml) reduced the biofilm formation capacity, which was statistically significant (p < 0.05). CONCLUSION: The results of this study showed that sub-MIC of Carvacrol has anti-biofilm effects in clinical A.baumannii colistin hetero-resistance isolates.

Investigating the effect of the inhibitory peptide on L.monocytogenes cell invasion: an in silico and in vitro study.

AIMS: L.monocytogenes monocytogenes is an omnipresent bacterium that causes a fatal food-borne illness, listeriosis. The connection of this bacterium to E-cadherin through internalin A plays a significant role in the internalization of the bacteria. In this study, this interaction has been investigated for the design of an inhibitory peptide. METHODS: The interaction of the proteins involved in the entry of bacteria was evaluated by molecular docking. According to their interactions, an inhibitory peptide was designed to bind to internalin A by server peptiderive. Its effects on L.monocytogenes invasion on the Caco-2 cell line and biofilm formation were also assessed. FINDINGS: Docking results showed that the peptide has a high affinity for binding to Internalin A. The synthesized peptide at a concentration of 64 µg/ml inhibited 80% of the invasion of L.monocytogenes into the Caco-2 cell line. Furthermore, the studied peptide at the highest concentration had a slight inhibitory effect on biofilm formation. CONCLUSION: These results reveal that short polypeptides can impede the invasion of target cells by L. monocytogenes in vitro and could be advantageous as restoring agents in vivo.

abkBA toxin-antitoxin system may act as antipersister modules in Acinetobacter baumannii clinical isolates.

Aim: Persistence cells comprise a subpopulation of bacteria that is resistant to treatment. In this study, the role of the toxin-antitoxin (TA) system in the formation of persistence cells of Acinetobacter baumannii isolates was investigated. Methods: After confirming all isolates, TA systems abkBA, mqsRA and higBA were identified. Persister cells were confirmed using the standard method. Real-time PCR was used to compare the expression of TA systems in isolates in persistence and normal states. Results: The abkAB system was present in all isolates; 4% of isolates formed persister cells. The expression level of the abkB gene in persistent isolates showed a sevenfold increase compared with nonpersistent isolates. Conclusion: The abkBA system is proposed as an antipersistence target in A. baumannii isolates.

Worldwide Prevalence of Colistin Resistance among Enterobacteriaceae: a Systematic Review and Meta-Analysis.

BACKGROUND: The aim of the present meta-analysis is to estimate the prevalence of colistin resistance among the Enterobacteriaceae family. METHODS: Articles from various databases (Medline, Scopus, Web of Science, and Embase) examining colistin resistance among Enterobacteriaceae in human, animal, and environmental specimens were searched from 2016 to 2021 using related keywords. The Cochran's Q-test and I2 were applied to evaluate heterogeneity and a random-effects model was used to assess the pooled prevalence. The meta-regression method was applied to determine heterogeneity among the studies. RESULTS: Of 5,145 articles, 60 articles with a sample size of 404,856 was included. The pooled estimate for prevalence of bacterial resistance were 9.13% (95% CI: 6.96 to 11.56; I-squared = 99.4%) in total, 8.34% (95% CI: 5.87 to 11.16; I-squared = 99.3%) for Klebsiella spp. subgroup and 3.44% (95% CI: 2.46 to 4.57; I-squared = 98.4%) for E. coli subgroup. The pooled prevalence for human and animal settings were 9.07% (95% CI: 6.77 to 11.67; I-squared = 99.3%) and 9.73% (95% CI: 484 to 16.02; I-squared = 99.4%), respectively. The continent (coefficient: 3.51; 95% CI: 0.08 to 6.94, p: 0.045) and bacterial type (coefficient: 0.03; 95% CI: 0.01 to 0.05 p: 0.042) had significant effects on heterogeneity among studies. CONCLUSION: The results of this study showed that the prevalence of colistin resistance in Enterobacteriaceae was similar between animals and humans, with the highest colistin resistance found in Klebsiella strains.

Development of innovative multi-epitope mRNA vaccine against Pseudomonas aeruginosa using in silico approaches.

The rising issue of antibiotic resistance has made treating Pseudomonas aeruginosa infections increasingly challenging. Therefore, vaccines have emerged as a viable alternative to antibiotics for preventing P. aeruginosa infections in susceptible individuals. With its superior accuracy, high efficiency in stimulating cellular and humoral immune responses, and low cost, mRNA vaccine technology is quickly replacing traditional methods. This study aimed to design a novel mRNA vaccine by using in silico approaches against P. aeruginosa. The research team identified five surface and antigenic proteins and selected their appropriate epitopes with immunoinformatic tools. These epitopes were then examined for toxicity, allergenicity and homology. The researchers also checked their presentation and identification by major histocompatibility complex cells and other immune cells through valuable tools like molecular docking. They subsequently modeled a multi-epitope protein and optimized it. The mRNA was analyzed in terms of structure and stability, after which the immune system's response against the new vaccine was simulated. The results indicated that the designed mRNA construct could be an effective and promising vaccine that requires laboratory and clinical trials.

Inhibitory Effects of Carvacrol on Biofilm Formation and Expression of Biofilm Related Genes in Clinical Isolates of Enterococcus faecalis.

BACKGROUND: Nowadays, novel antimicrobial strategies are being developed which focus on debilitating, rather than killing the microorganisms. In this regard, anti-biofilm therapy is one of the important ways to combat bacterial infections. Therefore, the aim of the current study was to evaluate the anti-biofilm activity of Carvacrol against E. faecalis by means of its effects on biofilm formation as well as on the gene expression levels of the two biofilm related genes, Epa and Esp. METHODS: A total of 40 clinical strains of E. faecalis were collected from three hospitals in Tehran, Iran during 2020. These isolates were confirmed by biochemical and genotypic methods. Antibacterial and anti-biofilm activity of Carvacrol essence were determined according the standard protocol. Finally, expression level of the biofilm related genes (Epa and Esp) were evaluated before and after the treatment with Carvacrol. RESULTS: A total of 14 isolates were considered as strong biofilm producers and were used for analysis. Carvacrol essence showed the best antibacterial activity at 2,500 µg/mL concentration against all the isolates, the biofilm formation capacity was decreased by Carvacrol essence, and it was statistically significant (p < 0.05). Expression levels of the Esp gene were decreased in 5 isolates while increased in 3 isolates following the Carvacrol treatment. Ex-pression levels of the EpaI gene was significantly decreased (p < 0.05) in 4 isolates following the Carvacrol treatment. CONCLUSIONS: In conclusion, the results presented in this study suggest that carvacrol extract exhibits significant antimicrobial and anti-biofilm properties against E. faecalis, even against vancomycin resistant isolates.

Clofazimine susceptibility testing of Mycobacterium avium complex and Mycobacterium abscessus: a meta-analysis study.

OBJECTIVES: The incidence of infections due to Mycobacterium avium complex (MAC) and Mycobacterium abscessus (MABS) is increasing worldwide. Current antimycobacterial agents are not sufficiently effective against nontuberculous mycobacteria (NTM) and there is a need for new drugs. This study aimed to estimate the overall in vitro activity of clofazimine (CFZ) against MAC and MABS clinical isolates. METHODS: We systematically searched four databases up to 1 March 2020 to identify relevant studies. Studies were included if they used the Clinical and Laboratory Standards Institute (CLSI) criteria for drug susceptibility testing (DST). We assessed the pooled in vitro CFZ resistance rate in MAC and MABS clinical isolates using a random- effects model. Sources of heterogeneity were evaluated using Cochran's Q and the I2 statistic. Potential for publication bias was explored using Begg's and Egger's tests. All analyses were conducted using Stata 14.0. RESULTS: A total of 20 publications (11 reports for MAC and 15 for MABS) were included. The pooled rates of in vitro resistance to CFZ in clinical isolates of MAC and MABS were 9.0% [95% confidence interval (CI) 3.0-17.0%] and 16.0% (95% CI 4.0-34.0%), respectively. There was no evidence of publication bias. CONCLUSION: This study reports the frequency of CFZ resistance in clinical isolates of MAC and MABS. According to the results, establishing accurate DST methods for detecting CFZ resistance, performing DST for all NTM isolates to provide effective treatment, and continuous monitoring of drug resistance are suggested for the prevention and control of CFZ-resistant NTM.

Role of Clofazimine in Treatment of Mycobacterium avium Complex.

Background: Non-tuberculous mycobacteria (NTM), specifically Mycobacterium avium complex (MAC), is an increasingly prevalent cause of pulmonary dysfunction. Clofazimine has been shown to be effective for the treatment of M. avium complex, but there were no published large-scale analyses comparing clofazimine to non-clofazimine regimens in MAC treatment. The objective of this large-scale meta-analysis was to evaluate patient characteristics and treatment outcomes of individuals diagnosed with MAC and treated with a clofazimine-based regimen. Methods: We used Pubmed/Medline, Embase, Web of Science, and the Cochrane Library to search for studies published from January 1, 1990 to February 9, 2020. Two reviewers (SSH and NY) extracted the data from all eligible studies and differences were resolved by consensus. Statistical analyses were performed with STATA (version 14, IC; Stata Corporation, College Station, TX, USA). Results: The pooled success treatment rate with 95% confidence intervals (CI) was assessed using random effect model. The estimated pooled treatment success rates were 56.8% in clofazimine and 67.9% in non-clofazimine groups. Notably, success rates were higher (58.7%) in treatment of HIV patients with disseminated infection. Conclusions: Treatment was more successful in the non-clofazimine group overall. However, HIV patients with disseminated infection had higher treatment response rates than non-HIV patients within the clofazimine group.

Corrigendum to "Traditional and Modern Cell Culture in Virus Diagnosis"[Osong Public Health Res Perspect 2016;7(2):77-82].

[This corrects the article on p. 77 in vol. 7, PMID: 27169004.].

Clonal Lineage Diversity, Antibiotic Resistance, and Virulence Determinants Among Methicillin-Resistant and Methicillin-Susceptible Staphylococcus aureus Isolated from Nurses at a Teaching Hospital in Ilam, Iran: Successful Nares Decolonization by Mupirocin.

BACKGROUND: Staphylococcus aureus is known to be responsible for nosocomial infections, and the typing method was useful in managing the reservoir of bacteria. The main aim of this study was to determine the prevalence of S. aureus in the nares and hands of nurses working in Imam Khomeini hospital, Ilam, Iran, as well as to determine the clonal relatedness, antimicrobial susceptibility profiles, different virulence, and resistance determinants among these isolates. The evolution of mupirocin activity in the eradication of methicillin-resistant S. aureus (MRSA) and methicillin-susceptible S. aureus (MSSA) colonization in the nares of the healthcare workers in Ilam, Iran, was also determined in this study. MATERIALS AND METHODS: In this cross-sectional study, 80 nurses, auxiliary nurses, and service workers from Imam Khomeini Hospital were enrolled. MRSA, antibiotic susceptibility, and virulence determinants were evaluated. Then, the isolates were subjected to pulsed field gel electrophoresis (PFGE) and Staphylococcal cassette chromosome mec typing. RESULTS: Our results demonstrated that 23% of isolates were MRSA. PFGE results demonstrated that pulsotypes A (3 out of 30; 10%) and J (3 out of 30; 10%), pulsotypes E (2 out of 30; 6.7%), M (2 out of 30; 6.7%), P(2 out of 30; 6.7%), and V (2 out of 30; 6.7%) were the most predominant pulsotypes, respectively. CONCLUSION: We cannot give conclusive suggestions about the correlation between nasal carriage and infections, but we suggest the monitoring of all healthcare workers annually, decontamination of their noses by using mupirocin and other antistaphylococcal agents, and also the washing of their hands at least every 2 h.

Association between biofilm production, adhesion genes and drugs resistance in different SCCmec types of methicillin resistant Staphylococcus aureus strains isolated from several major hospitals of Iran.

OBJECTIVES: The ability of bacteria to produce biofilm and adhesion makes them more resistant to antibiotics. The current study aims to evaluate the biofilm formation by Staphylococcus aureus and to determine the prevalence of adhesion genes, also their correlation with drug resistance. MATERIALS AND METHODS: A total of 96 MRSA were collected from hospitals of Iran's western provinces during 2012 to 2013. The presence of ica A, B, C, D, clfA, cna, fnbA, mecA genes were determined by PCR technique. Biofilm formation was studied by microtiter plate assay, the clonal relations of the strains were examined by SCCmec and Spa typing. RESULTS: The results demonstrated that 96 % of isolates were biofilm producers. The distributions of biofilm formation between isolates were 4.2%, 54.2%, 35.4% as high, moderate and weak, respectivelly. The highest biofilm production was observed from blood culture isolates. All virulent genes icaA,B, C, D, clfA, cna, fnbA were observed in moderate and weak biofilm formation isolates. Among high biofilm formation isolates, icaB and cna genes were not seen. Statistical analysis showed that there was a significant correlation between ica, fnbA and the biofilm production, but there was not a significant correlation between the type of samples and drug resistance, spa type and SCCmec type with biofilm production (P>0.05). Frequency of All virulent genes in type III SCCmec was higher than other types. CONCLUSION: The majority of MRSA isolates were biofilm producers and blood isolates ranked as the great biofilm producer. In these isolates ica D and fnbA genes are correlated with biofilm production.

Traditional and Modern Cell Culture in Virus Diagnosis.

Cell cultures are developed from tissue samples and then disaggregated by mechanical, chemical, and enzymatic methods to extract cells suitable for isolation of viruses. With the recent advances in technology, cell culture is considered a gold standard for virus isolation. This paper reviews the evolution of cell culture methods and demonstrates why cell culture is a preferred method for identification of viruses. In addition, the advantages and disadvantages of both traditional and modern cell culture methods for diagnosis of each type of virus are discussed. Detection of viruses by the novel cell culture methods is considered more accurate and sensitive. However, there is a need to include some more accurate methods such as molecular methods in cell culture for precise identification of viruses.

The Prevalence of ESBLs Producing Klebsiella pneumoniae Isolates in Some Major Hospitals, Iran.

Aims of this study were to investigate on antibiotic resistance and molecular epidemiology of K.pneumoniae producing ESBLs isolates of respiratory tract infections in some major hospitals in Iran. K.pneumonaie were obtained of patients with RTI. K. pneumoniae producing ESBLs detected by screening, confirming and PCR methods. During the 12-month period, a total of one hundred and thirteen of K.pneumoniae were found from RTI in three cities in different region of Iran which Sixty seven strains (59.2%) were ESBLs producer. In Ilam hospitals, seventeen strains (43.6%), in Milad hospital, thirty-seven strains (74%) and in Emam Reza hospital, thirteen strains (54.2%) were ESBLs producer. The findings showed that among sixty-seven K.pneumoniae producing ESBLs, Sixty-three strains (94%) were positive for blaSHV, eleven strains (16.4%) contained blaTEM and sixteen strains (23.9%) harbored blaCTX-M. Imipenem was found as an effectiveness antibiotic. In the current study, Majority of the ESBLs production had occurred in Milad hospital in Tehran (74%). In conclusion, spreading ESBL-producing strains is a concern, as it causes limitations to the antimicrobial agents for optimal treatment of patients.

Rapid identification of Iranian Acinetobacter baumannii strains by single PCR assay using BLA oxa-51 -like carbapenemase and evaluation of the antimicrobial resistance profiles of the isolates.

The rapid identification of relevant bacterial pathogens is of utmost importance in clinical settings. The aim of this study was to test a rapid identification technique for A. baumannii strains from Tehran Hospitals and to determine the antibiotic resistance profiles of the isolates. A hundred strains of Acinetobacter spp. grown from clinical specimens were identified as A. baumannii by conventional methods. Using PCR a bla OXA-51 -like gene was detected in all A. baumannii isolates but not in other species of acinetobacter. More than half of the isolates proved resistant to a variety of antibiotics by the disk diffusion technique. The rate of resistance to gentamicin, imipenem, ampicillin-sulbactam and amikacin was determined to be 45%, 53%, 62% and 62%, respectively. Moreover, most isolates (more than 90%) showed resistance to cephalosporins. This study shows that the demonstration of the bla OXA-51-like gene is a reliable and rapid way for the presumptive identification of A. baumannii and reveals that the rate of antibiotic resistance is high in Iranian A. baumannii isolates to a variety of antibiotics.