High Levels Of Antibiotic Resistance Research Articles

Multiplex PCR assay to detect high risk lineages of Salmonella Typhi and Paratyphi A.

Enteric fever infections remain a significant public health issue, with up to 20 million infections per year. Increasing rates of antibiotic resistant strains have rendered many first-line antibiotics potentially ineffective. Genotype 4.3.1 (H58) is the main circulating lineage of S. Typhi in many South Asian countries and is associated with high levels of antibiotic resistance. The emergence and spread of extensively drug resistant (XDR) typhoid strains has increased the need for a rapid molecular test to identify and track these high-risk lineages for surveillance and vaccine prioritisation. Current methods require samples to be cultured for several days, followed by DNA extraction and sequencing to determine the specific lineage. We designed and evaluated the performance of a new multiplex PCR assay, targeting S. Paratyphi A as well as the H58 and XDR lineages of S. Typhi on a collection of bacterial strains. Our assay was 100% specific for the identification of lineage specific S. Typhi and S. Paratyphi A, when tested with a mix of non-Typhi Salmonella and non-Salmonella strains. With additional testing on clinical and environmental samples, this assay will allow rapid lineage level detection of typhoid of clinical significance, at a significantly lower cost to whole-genome sequencing. To our knowledge, this is the first report of a SNP-based multiplex PCR assay for the detection of lineage specific serovars of Salmonella Typhi.

Bacteria associated with leaf-cutter ants drive natural antibiotic resistance in soil bacteria

AbstractActinobacteria that live mutualistically with leaf-cutter ants secrete antibiotics that may induce antibiotic resistance in nearby soil bacteria. We tested for the first time whether soil bacteria near and inside Atta cephalotes nests in Costa Rica show higher levels of antibiotic resistance than bacteria collected farther away. We collected soil samples 0 m to 50 m away from ant nests and grew bacteria from them on agar with paper discs treated with antibiotics of common veterinary use. As a proxy for antibiotic resistance, we measured the distance from the edge of each disc to the closest bacterial colonies. In general, resistance to oxytetracycline increased with proximity to leaf-cutter ant nests. Antibiotic resistance to oxytetracycline was also higher in samples collected inside the nest than in samples from the nest mound; not all antibiotics demonstrated the same trend. A preliminary exploratory morphological analysis suggests bacterial communities between 0 m and 50 m from ant nests were similar in diversity and abundance, indicating the pattern of antibiotic resistance described above may not be caused by differences in community composition. We conclude that actinobacteria living mutualistically with A. cephalotes drive natural antibiotic resistance to tetracycline in proximal bacterial communities.

The Frequency of Occurrence of Resistance and Genes Involved in the Process of Adhesion and Accumulation of Biofilm in Staphylococcus aureus Strains Isolated from Tracheostomy Tubes.

Background: Bacterial biofilm on the surface of tracheostomy tubes (TTs) is a potential reservoir of potentially pathogenic bacteria, including S. aureus. For this reason, our study aimed to investigate biofilm production in vitro and the presence of icaAD and MSCRAMM genes in clinical S. aureus strains derived from TTs, with respect to antibiotic resistance and genetic variability. Methods: The clonality of the S. aureus strains was analyzed by the PFGE method. The assessment of drug resistance was based on the EUCAST recommendations. The isolates were evaluated for biofilm production by the microtiter plate method and the slime-forming ability was tested on Congo red agar (CRA). The presence of icaAD genes was investigated by PCR and MSCRAMM genes were detected by multiplex PCR. Results: A total of 60 patients were enrolled in the study. One TT was obtained from each patient (n = 60). Twenty-one TTs (35%) were colonized with S. aureus. A total of 24 strains were isolated as 3 patients showed colonization with 2 SA clones (as confirmed by PFGE). PFGE showed twenty-two unique molecular profiles. Two isolates (8%) turned out to be MRSA, but 50% were resistant to chloramphenicol, 25% to erythromycin and 8% to clindamycin (two cMLSB and four iMLSB phenotypes were detected). The microtiter plate method with crystal violet confirmed that 96% of the strains were biofilm formers. Representative strains were visualized by SEM. All isolates had clfAB, fnbA, ebpS and icaAD. Different MSCRAMM gene combinations were observed. Conclusions: the present study showed that the S. aureus isolated from the TTs has a high diversity of genotypes, a high level of antibiotic resistance and ability to produce biofilm.

Scenario of Antibiotic Resistance in Pakistan: A Systematic Review

Background: A rise in antimicrobial resistance and a decrease in the discovery of novel antibiotics have occurred in recent years. Pakistan is a South Asian developing nation with a high level of antibiotic resistance, which poses a serious regional and global threat. Objective: To determine the situation of antimicrobial resistance in Pakistan Methodology: In the current study the literature was reviewed systematically according to the guidelines of PRISMA. Numerous databases like PUBMED, Embase, Medline and Google scholar were reviewed to determine the prevalence of antimicrobial resistance in Pakistan in the last 10 years from 2012 to 2022. For collection of duplicate data, a standardized extractive datasheet was used. All the data was entered in Microsoft Excel 2019. Results: In the current study, a total of 82 articles were included for final analysis. Multiple drug resistant bacteria were observed in 8 (9.76%) studies while extensive drug resistant bacteria were observed in 2 (2.44%) studies. High median resistance (95% CI) was shown by E. coli to first line antibiotics. Salmonella spp showed high median resistance to ciprofloxacin (90.5%). High median resistance was observed to majority of available antibiotics by Acinetobacter spp. Pseudomonas spp were highly susceptible (86.5%) to cefoperazone-sulbactam. MRSA S. aureus was reported in 13 studies and they were highly resistant to penicillin (98%), cefoxitin (83%) and levofloxacin (80%). Conclusion: Our study concludes that antimicrobial resistance pattern of common pathogens was high against commonly prescribed antibiotics. Regularization of monitoring practices, regular local and national monitoring, molecular investigations, and particular efforts to address the risk associated with the rise in antibiotic resistance are required to combat this issue. Keywords: Antibiotic resistance; Multiple drug resistance, Extensive drug resistance

A high level of antibiotic resistance in Klebsiella and Aeromonas isolates from street water sold in Mozambique, associated with the prevalence of extended-spectrum and AmpC ß-lactamases

This study aims to evaluate the resistance profile and the prevalence of antibiotic resistance genes in 30 isolates of Klebsiella spp. and Aeromonas spp. recovered from water sold in the streets of Maputo. Susceptibility profiles to 15 antibiotics were performed according to Clinical Laboratory Standard Institute guidelines with antibiotic disks on Mueller-Hinton agar plates. Multiplex PCRs were performed targeting 10 ß-lactamase genes, five ESBL (blaTEM-variants, blaOXA-variants, BlaSHV-variants, MCTX-M Group 1 and Group 9 variants) and five AmpC (ACC variants, FOX variants, MOX variants, CIT variants and DHA variants). The results showed a high prevalence of Klebsiella resistance to ß-lactam antibiotics, such as amoxicillin/clavulanic acid (62.5%), amoxicillin (56.3%), ampicillin (50%), cefoxitin (43.8%), and cefotaxime (43.8%). Aeromonas showed resistance to cefoxitin and ampicillin (71.4%), amoxicillin/clavulanic acid (57.1%) and imipenem (42.9%). ESBL blaOXA -variants, blaSVH -variants, MCTX-M Group 1 variants, and MCTX-M Group 9 variants were the most prevalent b-lactam genes, followed by the b-lactams AmpC, ACC variants and FOX variants. It is extremely important to improve waterborne disease control strategies, especially in terms of public awareness of the potential health implications of multidrug-resistant strains of Klebsiella and Aeromonas, which are often neglected.

The Efficacy of Using Combination Therapy against Multi-Drug and Extensively Drug-Resistant Pseudomonas aeruginosa in Clinical Settings.

Pseudomonas aeruginosa is a Gram-negative bacterium which is capable of developing a high level of antibiotic resistance. It has been placed on the WHO’s critical priority pathogen list and it is commonly found in ventilator-associated pneumonia infections, blood stream infections and other largely hospital-acquired illnesses. These infections are difficult to effectively treat due to their increasing antibiotic resistance and as such patients are often treated with antibiotic combination regimens. Methods: We conducted a systematic search with screening criteria using the Ovid search engine and the Embase, Ovid Medline, and APA PsycInfo databases. Results: It was found that in many cases the combination therapies were able to match or outperform the monotherapies and none performed noticeably worse than the monotherapies. However, the clinical studies were mostly small, only a few were prospective randomized clinical trials and statistical significance was lacking. Conclusions: It was concluded that combination therapies have a place in the treatment of these highly resistant bacteria and, in some cases, there is some evidence to suggest that they provide a more effective treatment than monotherapies.

Human Cryptic Host Defence Peptide GVF27 Exhibits Anti-Infective Properties against Biofilm Forming Members of the Burkholderia cepacia Complex.

Therapeutic solutions to counter Burkholderia cepacia complex (Bcc) bacteria are challenging due to their intrinsically high level of antibiotic resistance. Bcc organisms display a variety of potential virulence factors, have a distinct lipopolysaccharide naturally implicated in antimicrobial resistance. and are able to form biofilms, which may further protect them from both host defence peptides (HDPs) and antibiotics. Here, we report the promising anti-biofilm and immunomodulatory activities of human HDP GVF27 on two of the most clinically relevant Bcc members, Burkholderia multivorans and Burkholderia cenocepacia. The effects of synthetic and labelled GVF27 were tested on B. cenocepacia and B. multivorans biofilms, at three different stages of formation, by confocal laser scanning microscopy (CLSM). Assays on bacterial cultures and on human monocytes challenged with B. cenocepacia LPS were also performed. GVF27 exerts, at different stages of formation, anti-biofilm effects towards both Bcc strains, a significant propensity to function in combination with ciprofloxacin, a relevant affinity for LPSs isolated from B. cenocepacia as well as a good propensity to mitigate the release of pro-inflammatory cytokines in human cells pre-treated with the same endotoxin. Overall, all these findings contribute to the elucidation of the main features that a good therapeutic agent directed against these extremely leathery biofilm-forming bacteria should possess.

Mcr Genes Conferring Colistin Resistance in Enterobacterales; a Five Year Overview.

The present review aims to study and detect the global emergence of mcr genes in E. coli, K. pneumoniae and Salmonella spp., isolates from human specimens over the last six years. Nowadays the rise of multidrug-resistant superbugs has made essential the return of drugs that were previously abandoned. A clear example is colistin, which acts against multidrug - resistant gram - negative pathogens, including Enterobacterales. Colistin resistance is an unfortunate fact, with the emergence of mcr genes conferring resistance to colistin in Enterobacterales posing the most recent threat. Literature about mcr genes and their spread in E. coli, K. pneumoniae and Salmonella spp. is cited, focusing on the emergence of mcr genes in human specimens since 2015. The data were taken from the PubMed and Scopus databases. It seems that the mcr-1 gene continues to be the protagonist among the three species. E. coli is the dominant species harbouring mcr genes. Moreover, plasmid - mediated colistin resistance is also conferred upon other species that carry different genes resistant to antibiotics. There are only scarse reports on human Salmonella spp isolates harbouring mcr genes. Finally, the emergence of the mcr-9 gene in all of them is quite remarkable. CONCLUSION: Plasmid - mediated colistin resistance in Enterobacterales is a global issue and has been worsening over the years. The continuous mutations of mcr gene subtypes underline the need for better surveillance, constant investigation and wise use of colistin, especially in countries with high levels of antibiotic resistance.

Molecular characterization of Staphylococcus aureus ST6 and ST7 isolates from food-borne illness outbreaks

Objective: To analyze the Staphylococcal enterotoxins, Staphylococcal enterotoxin genes, drug resistance and molecular typing of 41 Staphylococcus aureus isolates from 2 food-borne illness outbreaks on 21 August and 27 September 2020 in Guangzhou. Methods: A total of 41 Staphylococcus aureus isolates from 2 outbreaks were analyzed by multilocus sequence typing (MLST) and spa typing. The Staphylococcal enterotoxins typing and the Staphylococcal enterotoxin genes of the isolates were analyzed by ELISA and PCR, respectively. The antimicrobial susceptibility of the isolates was performed by disc diffusion. 21 Staphylococcus aureus isolates were characterized using whole genome sequencing (WGS). Based on the whole genome single nucleotide polymorphism (SNP), the phylogenetic tree was constructed by Snippy. Results: 41 Staphylococcus aureus isolates were divided into 2 types by MLST and spa typing: ST6-t701 and ST7-t091. 2 ST7-t091 isolates were identified as methicillin-resistant Staphylococcus aureus (MRSA). 25 ST7-t091 isolates and 14 ST6-t701 isolates were methicillin-sensitive Staphylococcus aureus (MSSA), and were resistant to 7 and 6 antibiotics, respectively. All isolates were positive for sea by PCR. WGS revealed all 21 isolates carried scn, sak, sea, hla, hld, hlgA, hlgB, hlgC, lukD virulence genes. The results showed the isolates contained an immune evasion cluster type D which located in bacteriophage ϕSa3. The SNP phylogenetic tree showed 2 MRSA ST7-t091 were constituted a separate clade from the 12 MSSA ST7-t091 isolates and 7 ST6-t701 isolates showed high similarity to each other. Conclusion: Base on the results of phylogenetic analysis, the 2 food-borne illness outbreaks occurred on 21 August and 27 September 2020 are caused by the combination of the MRSA ST7-t091 strain and the MSSA ST7-t091 strain, and the MSSA ST6-t701 strain, respectively. All isolates have high level of antibiotic resistance and carry high virulent genes.

Activity of Sulbactam-Durlobactam and Comparators Against a National Collection of Carbapenem-Resistant Acinetobacter baumannii Isolates From Greece.

Background Acinetobacter baumannii is a leading cause of healthcare-associated infections worldwide, due to both its persistence in the hospital setting and ability to acquire high levels of antibiotic resistance. Carbapenem-resistant A. baumannii isolates (CRAB) limit the activity of current antimicrobial regimens and new alternatives or adjuncts to traditional antibiotics are urgently needed. Durlobactam is a novel broad-spectrum inhibitor of serine-type β-lactamases that restores sulbactam (SUL) activity against A. baumannii. The sulbactam-durlobactam (SD) combination has recently completed Phase 3 testing in the global ATTACK trial.ObjectivesThe aim of this study is to evaluate the in vitro activity of SD versus comparators against a representative nationwide collection of CRAB isolates.MethodsOne hundred ninety CRAB isolates were collected from clinical samples of patients hospitalized in 11 hospitals throughout Greece during 2015. In vitro activities of SD and comparators (SUL alone, amikacin, minocycline, imipenem, meropenem, colistin, SD and imipenem combined with SD) were determined by broth microdilution.ResultsDurlobactam restored sulbactam activity against the majority of the strains tested, with SD exhibiting the lowest MIC90 (8 μg/ml) relative to the other single comparators tested; 87.9% of the isolates had SD MICs ≤4/4 µg/ml. The most active comparator was colistin (MIC90 = 16 μg/ml). The addition of imipenem further lowered the MIC90 of SD by one two-fold dilution.ConclusionsThis study demonstrated the potential utility of SD for the treatment of infections caused by A. baumannii. If its clinical efficacy is confirmed, SD may be an important therapeutic option for CRAB infections.

The Role of ZntA in Klebsiella pneumoniae Zinc Homeostasis.

ABSTRACTKlebsiella pneumoniae is an opportunistic Gram-negative pathogen that is a leading cause of healthcare-associated infections, including pneumonia, urinary tract infections, and sepsis. Essential to the colonization and infection by K. pneumoniae is the acquisition of nutrients, such as the transition metal ion zinc. Zinc has crucial structural and catalytic roles in the proteome of all organisms. Nevertheless, in excess, it has the potential to mediate significant toxicity by dysregulating the homeostasis of other transition elements, disrupting enzymatic processes, and perturbing metalloprotein cofactor acquisition. Here, we sought to elucidate the zinc detoxification mechanisms of K. pneumoniae, which remain poorly defined. Using the representative K. pneumoniae AJ218 strain, we showed that the P-type ATPase, ZntA, which is upregulated in response to cellular zinc stress, was the primary zinc efflux pathway. Deletion of zntA rendered K. pneumoniae AJ218 highly susceptible to exogenous zinc stress and manifested as an impaired growth phenotype and increased cellular accumulation of the metal. Loss of zntA also increased sensitivity to cadmium stress, indicating a role for this efflux pathway in cadmium resistance. Disruption of zinc homeostasis in the K. pneumoniae AJ218 ΔzntA strain also impacted manganese and iron homeostasis and was associated with increased production of biofilm. Collectively, this work showed the critical role of ZntA in K. pneumoniae zinc tolerance and provided a foundation for further studies on zinc homeostasis and the future development of novel antimicrobials to target this pathway.IMPORTANCE Klebsiella pneumoniae is a leading cause of healthcare-associated infections, including pneumonia, urinary tract infections, and sepsis. Treatment of K. pneumoniae infections is becoming increasingly challenging due to high levels of antibiotic resistance and the rising prevalence of carbapenem-resistant, extended-spectrum β-lactamases producing strains. Zinc is essential to the colonization and infection by many bacterial pathogens but toxic in excess. This work described the first dissection of the pathways associated with resisting extracellular zinc stress in K. pneumoniae. This study revealed that the P-type ATPase ZntA was highly upregulated in response to exogenous zinc stress and played a major role in maintaining bacterial metal homeostasis. Knowledge of how this major bacterial pathogen resists zinc stress provided a foundation for antimicrobial development studies to target and abrogate their essential function.

Evaluation of the bioresistance profile of enterobacteria isolated from faeces of children with diarrhoea in the town of Koula-Moutou, Gabon: prospective study.

the emergence and expansion of multidrug resistance in Enterobacteriaceae responsible for various infections are increasing in the world. This study was designed to determine the phenotypic profiles of the resistance of enterobacteria strains isolated from the faeces of children with diarrhoeal diseases and to classify them according to the type of resistance. screening was carried out on 98 isolates divided into 2 groups: opportunistic pathogens and strict enteropathogens. Their sensitivity to 13 antibiotics was evaluated by the Mueller Hinton agar medium diffusion method. a strong resistance to different classes of β-lactams was found in the strains, 74.0% (n=45) and 83.3% (n=31) for opportunists and enteropathogens, respectively. These strains were completely resistant to doxycycline and erythromycin (100%; n=98) for both types of bacteria. Opportunists and enteropathogens were 95.1% (n=58) and 94.6% (n=35) resistant to gentamicin, and 31.1% (n=19) and 35.1% (n=13) resistant to chloramphenicol, respectively. Similarly, the total resistance of strains was observed with ofloxacin and amounted to 98.4% (n=60) and 96.7% with levofloxacin and norfloxacin, respectively. The analysis of β-lactam resistance phenotypes revealed a dominance of the carbapenemase-producing strains (28.6%; n=28). However, 24.3% (n=9) of enteropathogens were pan-resistant versus 19.7% (n=12) for opportunists. the results of this study indicate a worryingly high level of antibiotic resistance in enterobacteria which might tend towards total resistance.

INVESTIGATION OF TIGECYCLINE SUSCEPTIBILITY OF MULTIDRUG-RESISTANT ACINETOBACTER BAUMANNII ISOLATES BY DISC DIFFUSION, AGAR GRADIENT AND BROTH MICRODILUTION TESTS.

The use of tigecycline is becoming increasingly important because of the high levels of antibiotic resistance in Acinetobacter baumannii (A. baumannii) isolates. In this prospective study, multidrug-resistant A. baumannii isolates were obtained from various tissue and fluid samples of patients admitted to or treated at various departments and tested in Laboratory of Microbiology, Duzce University Medical Faculty between January 2013 and December 2015. Tigecycline resistance in multidrug-resistant A. baumannii isolates were analyzed using the disc diffusion test (DDT), agar gradient test (AGT), and gold standard test [broth microdilution test (BMT)]. A. baumannii isolates resistant to multiple drugs were included in the study (N=94). Using the BMT method, 89 (95%), 4 (4%) and 1 (1%) A. baumannii isolates were determined as tigecycline susceptible, intermediate and resistant isolates, respectively. Using the Food and Drug Administration criteria, the rates of major error (ME), minor error (mE) and categorical agreement (CA) for DDT were 26%, 67% and 9%, respectively. In contrast, for AGT, the rates of ME, mE and CA were 0%, 4%, 95%, respectively. Tigecycline resistance as assessed by BMT showed no increase between 2013 and 2015. Accordingly, isolates found to be resistant or intermediate by DDT should be confirmed by BMT. Due to the ease of application, AGT is a safe method of detecting susceptibility.

The Fate of Foodborne Pathogens in Manure Treated Soil.

The aim of this review was to provide an update on the complex relationship between manure application, altered pathogen levels and antibiotic resistance. This is necessary to protect health and improve the sustainability of this major farming practice in agricultural systems based on high levels of manure production. It is important to consider soil health in relation to environment and land management practices in the context of the soil microflora and the introduction of pathogens on the health of the soil microbiome. Viable pathogens in manure spread on agricultural land may be distributed by leaching, surface run-off, water source contamination and contaminated crop removal. Thus it is important to understand how multiple pathogens can persist in manures and on soil at farm-scale and how crops produced under these conditions could be a potential transfer route for zoonotic pathogens. The management of pathogen load within livestock manure is a potential mechanism for the reduction and prevention of outbreaks infection with Escherichia coli, Listeria Salmonella, and Campylobacter. The ability of Campylobacter, E. coli, Listeria and Salmonella to combat environmental stress coupled with their survival on food crops and vegetables post-harvest emphasizes the need for further study of these pathogens along with the emerging pathogen Providencia given its link to disease in the immunocompromised and its’ high levels of antibiotic resistance. The management of pathogen load within livestock manure has been widely recognized as a potential mechanism for the reduction and prevention of outbreaks infection but any studies undertaken should be considered as region specific due to the variable nature of the factors influencing pathogen content and survival in manures and soil. Mediocre soils that require nutrients could be one template for research on manure inputs and their influence on soil health and on pathogen survival on grassland and in food crops.

Identification of multi-antibiotic resistant bacteria isolated from Vietnamese climbing perch (Anabas testudineus) on fish farms in Ho Chi Minh City, Vietnam

Climbing perch (Anabas testudineus) is considered a cultured species with high economic value and is widely distributed in several Asian countries, including Vietnam. In climbing perch aquaculture, antibiotic and antibacterial agents have been overused, leading to the emergence of antibiotic-resistant bacteria or genes in the aquatic environment as well as in the seafood products. These antibiotic-resistant bacteria and/or genes can then be spread to human through the food chain, generating detrimental consequences for public health and environment. In this study, 14 bacterial strains were isolated from Vietnamese climbing perch collected from various fish farms in Ho Chi Minh City which possess abnormal symptoms on the outside and in their organs. These bacterial strains were then subjected to several biochemical tests to characterize their colony morphologies. Besides, gene sequencing was performed and as compared to the 16S rRNA, 4 bacterial strains were identified with a high percentage in similarity, including Pseudomonas aeruginosa (99.7%), Aeromonas caviae (99.93%), Kosakonia sacchari (100%), and Edwardsiella ictaluri (99.44%). Additionally, these bacteria showed high levels of antibiotic resistance against beta-lactam, tetracycline, and others. Taken together, these results showed that the four identified bacterial strains isolated from Vietnamese climbing perch circulated in fish farms in Ho Chi Minh City. Their current status of multi-antibiotic resistance indicates further alternative sources of antimicrobial agents, for example, natural compounds, to overcome antibiotic resistance. It is then crucial to identify fish diseases, the associated pathogens, and the new alternatives for antibiotics to protect aquatic ecosystems and water resources.

A scoping review protocol to identify clinical signs, symptoms and biomarkers indicative of biofilm presence in chronic wounds

Introduction: Wound healing is characterised by haemostatic, inflammatory, proliferative and remodelling phases. In the presence of comorbidities such as diabetes, healing can stall and chronic wounds may result. Infection is detrimental to these wounds and associated with poor outcomes. Wounds are contaminated with microbes and debris, and factors such as host resistance, bacterial virulence, species synergy and bioburden determine whether a wound will deteriorate to critically colonised/infected states. Biofilms are sessile microbial communities, exhibiting high-level antibiotic tolerance and resistance to host defences. Biofilm in critically colonised wounds can contribute to delayed healing. Little is known about clinical presentation and diagnosis of wound biofilms. Objective: To identify from the literature clinical signs, symptoms and biomarkers that may indicate biofilm presence in chronic wounds. Methods: This review will be guided by the Preferred Reporting Items for Systematic Reviews extension for Scoping Reviews (PRISMA-ScR), and the Joanna Briggs Institute Manual for Evidence Synthesis. Studies of any design in any language recruiting adult patients with venous, diabetic, pressure or mixed arterial-venous ulcers and reporting data on clinical signs/symptoms of biofilm are eligible. Searches of Medline, Embase, CINAHL, Cochrane Central and BASE will be conducted from inception to present. Reference scanning and contact with content experts will be employed. Title/abstract screening and full text selection will be executed by two reviewers independently. Discrepancies will be resolved by discussion between reviewers or through third party intervention. Data will be extracted by a single reviewer and verified by a second. Clinical signs and symptoms data will be presented in terms of study design, setting and participant demographic data. Discussion: Understanding biofilm impact on chronic wounds is inconsistent and based largely on in vitro research. This work will consolidate clinical signs, symptoms and biomarkers of biofilm in chronic wounds reported in the literature.

Phenotypic and molecular evaluation of biofilm formation in Klebsiella pneumoniae carbapenemase (KPC) isolates obtained from a hospital of Pelotas, RS, Brazil.

Introduction. A significant cause of mortality in the intensive care unit (ICU) is multidrug-resistant (MDR) Gram-negative bacteria, such as Klebsiella pneumoniae carbapenemase (KPC). Biofilm production is a key factor in KPC colonization and persistence in the host, making the treatment difficult.Gap Statement. The aim of this study was to evaluate the antibiotic resistance, molecular and phenotypic biofilm profiles of 12 KPC isolates associated with nosocomial infection in a hospital in Pelotas, Rio Grande do Sul, Brazil.Methodology. Clinical isolates were obtained from different sources, identified and characterized by antibiotic resistance and carbapenemase synthesis following the Clinical and Laboratory Standards Institute (CLSI) guidelines. Polymerase chain reaction (PCR) was used to evaluate the presence of carbapenemase (blaKPC) and biofilm formation-associated genes (fimA, fimH, rmpA, ecpA, mrkD and wabG). Additionally, phenotypic evaluation of in vitro biofilm formation capacity was evaluated by Congo red agar (CRA) assay and the crystal violet staining method.Results. The 12 isolates evaluated in this study presented the blaKPC gene and were positive for synthesizing carbapenemases in vitro. In the carbapenem class, 83.3 % isolates were resistant and 16.7 % intermediately resistant to imipenem and meropenem. Molecular analyses found that the fimA and wabG genes were detected in 75 % of isolates, while fimH and ecpA were detected in 42 % and mrkD were detected in 8.3 % (1). The CRA assay demonstrated that all isolates were slime producers and 91.7 % (11) of isolates were classified as strong and 8.3 % (1) as moderate biofilm producers by the crystal violet staining method. The optical density (OD540nm) for strong biofilm formers ranged from 0.80±0.05 to 2.47±0.28 and was 0.55±0.12 for moderate biofilm formers.Conclusion. Our study revealed a high level of antibiotic resistance and biofilm formation in KPC isolates obtained from a hospital in Pelotas, RS, Brazil.

Bacterial uropathogens and burden of antimicrobial resistance pattern in urine specimens referred to Ethiopian Public Health Institute.

Urinary tract infections (UTIs) are the leading causes of morbidity in the general population, and is the second most common infectious disease after respiratory infections. Appropriate antibiotic therapy is essential to achieving good therapeutic results. Therefore, the purpose of this study was to investigate the profile of pathogens cultured from urinary tract infections and to determine their resistance profiles to commonly prescribed antibiotics. A cross-sectional study was carried out at the National Referral Laboratory of the Ethiopian Institute of Public Health from January 2017 to December 2018. All positive cultures were characterized by colony morphology, Gram stain, and standard biochemical tests. The antimicrobial susceptibility test of the isolate was performed using the Kirby- Bauer disk diffusion test on Muller-Hinton agar. In addition, bacterial identification, antimicrobial susceptibility testing and phenotypic detection of MDR were performed with VITEK 2 Compact according to the manufacturer's instructions. Out of 1012 cultured urine specimens, 325 (32.1%) was showed significant bacteriuria. The overall prevalence of UTIs was 325(32.1%) and the highest prevalence rate was obtained from 21-30 years age group 73(22.5%). Among UTIs patients, 583(57.6%) were females and 429(42.4%) were males. The UTIs of 179 (55%) women is relatively higher than that of men 146 (45%). Among 325 isolates, Gram-negative bacteria (GNB) appeared more frequently 252 (51.7%) than Gram-positive bacteria 63 (19.4%). In GNB, E. coli 168(66.7%), Klebsiella species 32(12.7%), and Enterobacter species 13 (5.2%) were dominated isolates whereas in GPB accounted for coagulase-negative staphylococcus (CoNS) 33(52.4%), Enterococcus species 16(25.4%), and Staphylococcus aureus 10(15.9%). Major of the isolates showed high levels of antibiotic resistance to commonly prescribed antimicrobials. Imipenem, Amikacin, and Nitrofurantoin were the most sensitive antibiotics for Gram-negative isolates while Nitrofurantoin, clindamycin, and Gentamycin were effective against gram-positive uropathogens. Overall, 156/256(60.9%), 56/256(22.4%), 10/256(4%) of gram-negative isolates were MDR, XDR, and PDR respectively while among the GPB isolates, 34/63(53.1%), 10/63(15.8%), and 1/63(1.6%) were MDR, XDR, and PDR isolates respectively. Among the tested bacterial strains, 190/319 (59.5%) were MDR, 66/319 (20.7%) strains were XDR, and 11/319 (3.45%) were PDR isolated. The prevalence of urinary tract infection was high, and Gram-negative organisms were the most common causes of UTIs in this study. It was found that the resistance to commonly used antibiotics is very high. Early detection and close monitoring of MDR, XDR, or even PDR bacterial strains must be started by all clinical microbiology laboratories to reduce the menace of antimicrobial resistance that is now a global problem.

Microbiota of the lower respiratory tract in community-acquired pneumonia, including cases associated with SARS-CoV-2

Introduction. Many aspects of the pathogenesis and pathomorphology of pneumonia associated with novel coronavirus require a comprehensive study using modern diagnostic methods.The aim of the study was to study the microbiota of the lower respiratory tract in community-acquired pneumonia associated with SARS-CoV-2, to assess the antibiotic and phage resistance of circulating strains of microorganisms.Materials and methods. The analysis of biosamples from 486 patients undergoing inpatient treatment in five mono-hospitals in Tyumen and Tyumen region with a diagnosis of moderate and severe community-acquired pneumonia was carried out. In almost 90% of cases patients received oxygen therapy, about 8% of patients were connected to ventilators. The inoculation of the cultures with clinical samples was carried out for six months (from April to October 2020). The isolated bacterial strains were identified by mass spectrometry. The resistance to antimicrobial drugs and bacteriophages was assessed for identified isolated.Results. Gram-positive cocci, mainly opportunistic microorganisms of the genus Streptococcus and Candida fungi predominated in the microbiota of the lower respiratory tract of patients diagnosed with community-acquired pneumonia associated with SARS-CoV-2. At the same time, bacteria of the Enterobacteriaceae family and nonfermenting gram-negative bacteria were less common compared to patients without coronavirus infection. In the structure of pathogens, the leading position was occupied by the bacteria K. pneumoniae and Acinetobacter spp. The analysis of the sensitivity of microorganisms to antimicrobial drugs showed the highest resistance rates in strains of Acinetobacter spp., Enterococcus spp., Coagulase-negative Staphylococcus. It has been established that in the group of patients with community-acquired pneumonia associated with SARS-CoV-2, the risk of infection with Streptococcus spp. with high level of antibiotic resistance was 1.5 times higher, and taking into account the 95% confidence interval, the value of this indicator ranged from 1.1 to 2.1 times.Conclusion. The data obtained indicate that the microbiota of the lower respiratory tract in community-acquired pneumonia associated with SARS-CoV-2 is represented mainly by bacteria of the genus Streptococcus, which have a high level of resistance to antimicrobial drugs.

Identification of Escherichia coli strains in the vaginal cultures of healthy women and their patterns of antibiotic resistance.

Background: Bacterial vaginosis is the most common cause of vaginitis in women of childbearing age, and it predominantly affects young sexually active women. Escherichia coli is one of the most common bacteria found in the genital tract of non-pregnant (9–28%) and pregnant women (24–31%). E. coli strains can colonize the vaginal and endocervical regions in pregnant women, and may lead to the development of urinary tract, intra-amniotic or puerperal infections. Aim of the study: Isolation and identification of the antibiotic resistance patterns of extended spectrum beta-lactamase (ESBL)-producing and non-producing E. coli in the vaginal cultures of healthy women. Material and methods: Vaginal samples were taken from 55 healthy women. For the bacterial identification and resistance patterns, automated equipment from Beckman Coulter was used. Phenotypic techniques were used to confirm the presence or absence of ESBL. Results: Fifty-five cultures developed E. coli, with the rest of the strains corresponding to different bacteria. Of the 55 E. coli cultures, 35 (63.63%) were ESBL-producing and 20 (36.36%) did not produce ESBL. There was an 80% resistance to penicillin, and a 76.4% and 65.5% resistance to the first and fourth generation cephalosporins, respectively. A 45.5% resistance was observed for the fluoroquinolones, 52.7% for trimethoprim/ sulfamethoxazole, and 100% sensitivity to carbapenemics and amikacin. Conclusions: A large presence of vaginal ESBL-producing E. coli was observed in healthy women, which increases the risk of therapeutic failure due to high levels of antibiotic resistance.