Antimicrobial Resistance Patterns in Patients with Vaginal Discharge: A 2019-2022 Analysis at the National Health Laboratory in Eritrea.

Background: Antimicrobial resistance poses a significant global threat to the treatment of bacterial infections, particularly in low- and middle-income regions such as Africa. This study is aimed at analyzing antimicrobial resistance patterns in vaginal swab samples from patients at the National Health Laboratory from 2019 to 2022. Methods: This retrospective study examined patient records from vaginal swab analyses performed at the National Health Laboratory between January 1, 2019, and December 31, 2022. Ethical approval was obtained from the Ministry of Health Research Ethical Approval and Clearance Committee on 15/02/2023. Results: Of the 622 samples, 83% underwent microbial isolation and identification. Citrobacter spp. exhibited high resistance (>43%) to antibiotics such as cephalexin, ceftazidime, nalidixic acid, ampicillin, gentamicin, and tetracycline. E. coli showed resistance rates of more than 50% to ampicillin, trimethoprim-sulfamethoxazole, and tetracycline. Klebsiella spp. and Proteus spp. exhibited resistance rates that exceeded 47% to specific antibiotics. Gram-positive bacteria have resistance rates of more than 49% with ampicillin, trimethoprim-sulfamethoxazole, tetracycline, oxacillin, vancomycin, and penicillin G. In particular, S. aureus demonstrated no resistance to rifampicin or clindamycin, while Streptococcus spp. showed 100% resistance to rifampicin and vancomycin. Several species, including Proteus species, Streptococcus spp., S. aureus, and Klebsiella spp. exhibited multidrug resistance. Conclusion: Most gram-negative bacteria displayed higher resistance of >45% to ampicillin, trimethoprim-sulfamethoxazole, and tetracycline. Among gram-positive bacteria, a higher resistance rate with ampicillin, trimethoprim-sulfamethoxazole, tetracycline, oxacillin, vancomycin, and penicillin G was recorded. S. aureus showed no resistance to rifampicin and clindamycin, and Strep. spp. indicated 100% resistance to rifampicin and vancomycin. This study highlights critical gaps and areas for further exploration. Expanding the spectrum of antibiotics tested and investigating underlying multidrug resistance mechanisms would provide a more comprehensive understanding of resistance patterns.

A penicillinase-modified poly(N-isopropylacrylamide-co-acrylamide) smart hydrogel biosensor with superior recyclability for sensitive and colorimetric detection of penicillin G.

Antibiotics are widely used for treating bacterial infections. However, excessive or improper use of antibiotics can pose a serious threat to human health and water environments, and thus, developing cost-effective, portable and effective strategies to analyze and detect antibiotics is highly desired. Herein, we reported a responsive photonic hydrogel (RPH)-based optical biosensor (PPNAH) with superior recyclability for sensitive and colorimetric determination of a typical ß-lactam antibiotic penicillin G (PG) in water. This sensor was composed of poly(N-isopropylacrylamide-co-acrylamide) smart hydrogel with incorporated penicillinase and Fe3O4@SiO2 colloidal photonic crystals (CPCs). The sensor could translate PG concentration signals into changes in the diffraction wavelength and structural color of the hydrogel. It possessed high sensitivity and selectivity to PG and excellent detection performances for other two typical ß-lactam antibiotics. Most importantly, due to the unique thermosensitivity of the poly(N-isopropylacrylamide) moieties in the hydrogel, the PG-responded PPNAH sensor could be facilely regenerated via a simple physical method at least fifty times while without compromising its response performance. Besides, our sensor was suitable for monitoring the PG-contaminated environmental water and displayed satisfactory detection performances. Such a sensor possessed obvious advantages of superior recyclability, highly chemical stability, low production cost, easy fabrication, wide range of visual detection, simple and intuitive operation for PG detection, and environmental-friendliness, which holds great potential in sensitive and colorimetric detection of the PG residues in polluted water.

Evaluation of Antibiotic Penicillin G Activities Based on Electrochemical Measurement of a Tetrazolium Salt.

This study focused on the electrochemical properties of tetrazolium salts to develop a simple method for evaluating viable bacterial counts, which are indicators of drug susceptibility. Considering that the oxidized form of tetrazolium, which has excellent cell membrane permeability, changes to the insoluble reduced form formazan inside the cell, the number of viable cells was estimated based on the reduction current of the tetrazolium remaining in the bacterial suspension. Dissolved oxygen is an important component of bacterial activity. However, it interferes with the electrochemical response of tetrazolium. We estimated the number of viable bacteria in the suspension based on potential-selective current responses that were not affected by dissolved oxygen. Based on solubility, cell membrane permeability, and characteristic electrochemical properties of the tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium, we developed a method for rapidly measuring viable bacteria within one-fifth of the time required by conventional colorimetric methods for drug susceptibility testing.

Selective adsorption and identification of penicillin G sodium in milk by molecularly imprinted polymer doped carbon dot.

A carbon dot (CD) was prepared by o-phenylenediamine and water, which showed bright yellow fluorescence under ultraviolet light irradiation (λ = 580 nm), and verified good fluorescence quenching effect on penicillin G sodium (Png-Na). Using methacrylic acid as a functional monomer, ethylene glycol dimethacrylate as a crosslinker, and Png-Na as a template, a kind of composite microsphere combining CD and molecularly imprinted polymer (MIP) was synthesized by surface-initiated atomic transfer radical polymerization (SI-ATRP). For reasons of comparison, we also prepared MIP without CD and non-imprinted polymers (NIPs). Through static and dynamic adsorption experiments, the maximum adsorption capacity was 47.05 mg g-1 and the equilibrium time was 30 min. High-performance liquid chromatography (HPLC) was utilized to determine the content of Png-Na in the spiked milk samples. A sensitive, rapid, and simple method for determination of Png-Na in food samples was developed. The utilized approach enabled the quantification of Png-Na within the concentration range 20-1000 µg L-1 (with a limit of detection of 5 µg L-1). The recoveries achieved were in the range 93.3-98.2%, with a relative standard deviation of 1.2-4.2%. The results demonstrated that CD@MIP possessed the capability of specific adsorption and fluorescence detection of Png-Na, enabling simultaneous detection and enrichment of Png-Na in real samples.

Biodegradation of penicillin G sodium by Sphingobacterium sp. SQW1: Performance, degradation mechanism, and key enzymes.

Biodegradation is an efficient and cost-effective approach to remove residual penicillin G sodium (PGNa) from the environment. In this study, the effective PGNa-degrading strain SQW1 (Sphingobacterium sp.) was screened from contaminated soil using enrichment technique. The effects of critical operational parameters on PGNa degradation by strain SQW1 were systematically investigated, and these parameters were optimized by response surface methodology to maximize PGNa degradation. Comparative experiments found the extracellular enzyme to completely degrade PGNa within 60 min. Combined with whole genome sequencing of strain SQW1 and LC-MS analysis of degradation products, penicillin acylase and ß-lactamase were identified as critical enzymes for PGNa biodegradation. Moreover, three degradation pathways were postulated, including ß-lactam hydrolysis, penicillin acylase hydrolysis, decarboxylation, desulfurization, demethylation, oxidative dehydrogenation, hydroxyl reduction, and demethylation reactions. The toxicity of PGNa biodegradation intermediates was assessed using paper diffusion method, ECOSAR, and TEST software, which showed that the biodegradation products had low toxicity. This study is the first to describe PGNa-degrading bacteria and detailed degradation mechanisms, which will provide new insights into the PGNa biodegradation.

Synthesis and Synergistic Antimicrobial Efficacy of Covalent Conjugates Composed of Epsilon-Poly-l-lysine and Beta-Lactam Antibiotics.

The increasing severity of problems posed by drug-resistant pathogens has compelled researchers to explore innovative approaches for infection prevention. Among these strategies, conjugation methods stand out for their convenience and high efficacy. In this study, multiple covalent conjugates were synthesized, incorporating the natural antimicrobial peptide epsilon-poly-l-lysine (EPL) and two commonly used ß-lactam antibiotics: penicillin G or ampicillin. Enhanced antimicrobial efficacy against typical Gram-negative pathogens, along with faster kill kinetics compared to combination approaches, was demonstrated by the EPL-Ampicillin covalent conjugates. Their antimicrobial mechanism was also substantiated through SEM and fluorescence tests in this work, confirming the inheritance of membrane-disrupting properties from EPL. Furthermore, the excellent biocompatibility of the raw materials was reserved in the covalent conjugates. This simplified conjugation method holds promise for the development of infection therapeutic drugs and potentially restores the sensitivity of conventional antibiotics to drug-resistant pathogens by introducing membrane-disrupting mechanisms.

Early life exposure of infants to benzylpenicillin and gentamicin is associated with a persistent amplification of the gut resistome.

BACKGROUND: Infant gut microbiota is highly malleable, but the long-term longitudinal impact of antibiotic exposure in early life, together with the mode of delivery on infant gut microbiota and resistome, is not extensively studied. METHODS: Two hundred and eight samples from 45 infants collected from birth until 2 years of age over five time points (week 1, 4, 8, 24, year 2) were analysed. Based on shotgun metagenomics, the gut microbial composition and resistome profile were compared in the early life of infants divided into three groups: vaginal delivery/no-antibiotic in the first 4 days of life, C-section/no-antibiotic in the first 4 days of life, and C-section/antibiotic exposed in first 4 days of life. Gentamycin and benzylpenicillin were the most commonly administered antibiotics during this cohort's first week of life. RESULTS: Newborn gut microbial composition differed in all three groups, with higher diversity and stable composition seen at 2 years of age, compared to week 1. An increase in microbial diversity from week 1 to week 4 only in the C-section/antibiotic-exposed group reflects the effect of antibiotic use in the first 4 days of life, with a gradual increase thereafter. Overall, a relative abundance of Actinobacteria and Bacteroides was significantly higher in vaginal delivery/no-antibiotic while Proteobacteria was higher in C-section/antibiotic-exposed infants. Strains from species belonging to Bifidobacterium and Bacteroidetes were generally persistent colonisers, with Bifidobacterium breve and Bifidobacterium bifidum species being the major persistent colonisers in all three groups. Bacteroides persistence was dominant in the vaginal delivery/no-antibiotic group, with species Bacteroides ovatus and Phocaeicola vulgatus found to be persistent colonisers in the no-antibiotic groups. Most strains carrying antibiotic-resistance genes belonged to phyla Proteobacteria and Firmicutes, with the C-section/antibiotic-exposed group presenting a higher frequency of antibiotic-resistance genes (ARGs). CONCLUSION: These data show that antibiotic exposure has an immediate and persistent effect on the gut microbiome in early life. As such, the two antibiotics used in the study selected for strains (mainly Proteobacteria) which were multiple drug-resistant (MDR), presumably a reflection of their evolutionary lineage of historical exposures-leading to what can be an extensive and diverse resistome. Video Abstract.

A low-volume LC/MS method for highly sensitive monitoring of phenoxymethylpenicillin, benzylpenicillin, and probenecid in human serum.

Background: The optimization of antimicrobial dosing plays a crucial role in improving the likelihood of achieving therapeutic success while reducing the risks associated with toxicity and antimicrobial resistance. Probenecid has shown significant potential in enhancing the serum exposure of phenoxymethylpenicillin, thereby allowing for lower doses of phenoxymethylpenicillin to achieve similar pharmacokinetic/pharmacodynamic (PK/PD) targets. We developed a triple quadrupole liquid chromatography mass spectrometry (TQ LC/MS) analysis of, phenoxymethylpenicillin, benzylpenicillin and probenecid using benzylpenicillin-d7 and probenecid-d14 as IS in single low-volumes of human serum, with improved limit of quantification to support therapeutic drug monitoring. Methods: Sample clean-up was performed by protein precipitation using acetonitrile. Reverse phase chromatography was performed using TQ LC/MS. The mobile phase consisted of 55% methanol in water + 0.1% formic acid, with a flow rate of 0.4 mL min-1. Antibiotic stability was assessed at different temperatures. Results: Chromatographic separation was achieved within 2 minutes, allowing simultaneous measurement of phenoxymethylpenicillin, benzylpenicillin and probenecid in a single 15 µL blood sample. Validation indicated linearity over the range 0.0015-10 mg L-1, with accuracy of 96-102% and a LLOQ of 0.01 mg L-1. All drugs demonstrated good stability under different storage conditions. Conclusion: The developed method is simple, rapid, accurate and clinically applicable for the quantification of phenoxymethylpenicillin, benzylpenicillin and probenecid in tandem.

Antibiotic Resistance Profile of Clinical Strains of Enterococci from Secondary/Persistent Endodontic Infections: What do We Know? A Systematic Review of Clinical Studies.

INTRODUCTION: Enterococcus faecalis is the most common enterococcal species associated with infective endocarditis and 1 of the most commonly detected bacteria in cases of secondary/persistent endodontic infection (SPEI). Antimicrobial resistance is a global public health concern. This review aimed to answer the following research question: "Is there a change in the antibiotic resistance profile in clinical strains of E. faecalis over the years?". P (population) - patients with SPEI, I (intervention) -endodontic retreatment, C (comparison) -not included, O (outcome) - profile of Enterococci resistance and susceptibility to systemic antibiotics used. METHODS: Two authors independently performed study selection, data extraction, and risk of bias assessment. The literature search was conducted using the following electronic databases: PubMed, Scopus, EMBASE, Web of Science, and Medline. Clinical studies in which Enterococci strains were isolated to assess their antimicrobial resistance were included. RESULTS: Eleven clinical trials were included. Overall, E. faecalis isolated from teeth with SPEI presented an intermediate resistance to 16 antibiotics. In recent years, E. faecalis showed a little resistance to amoxicillin (without clavulanate) and benzylpenicillin. Erythromycin and rifampicin presented an increase in the intermediate-resistance status between the first and the last studies. E. faecium presented intermediate-resistance results. CONCLUSION: The most effective drugs remain the combination of amoxicillin and clavulanate, followed by amoxicillin and benzylpenicillin. In patients allergic to penicillin derivatives, moxifloxacin and azithromycin may be indicated with caution. The antibiotics with the highest pattern of resistance against E. faecalis are clindamycin, gentamicin, metronidazole, and rifampicin and are therefore, contraindicated in cases of SPEI. Very few clinical studies using a microbiological approach in teeth with endodontic failure have been carried out to improve the efficacy of prophylactic regimens. However, as bacteria periodically develop resistance to the main drugs used, regular studies should be carried out on the action of these drugs in infection control.

Rising Incidence of Syphilitic Uveitis-Related Hospitalizations in the US.

Importance: The number of syphilis cases continues to increase in the US every year since 2001 with a 74% increase observed since 2017. In addition, there remains a national shortage of injectable penicillin G. Despite the increase in reported cases, to the authors' knowledge, there has been no recent nationwide study investigating the trends in incidence of syphilitic uveitis. Objective: To evaluate the national and regional incidence of syphilitic uveitis-related hospitalizations in the US. Design, Setting, and Participants: This was a retrospective, cross-sectional study. The Nationwide Inpatient Sample was queried to identify all inpatient admissions with a diagnosis of syphilitic uveitis in the US between the years 2010 and 2019. Analyses were performed to determine baseline sociodemographic characteristics and identify national and regional trends in incidence. All patients hospitalized with a diagnosis of syphilis, uveitis, and/or syphilitic uveitis were eligible for inclusion. Statistical analysis of study data took place in June 2023. Exposure: Diagnosis of syphilis, uveitis, and/or syphilitic uveitis on inpatient admissions during the years 2010 to 2019 in the Nationwide Inpatient Sample. Main Outcome Measures: The primary outcome was to determine trends in the national and regional incidence of syphilitic uveitis-related hospitalizations in the US. Secondary outcome measures included sociodemographic characteristics of patients with syphilitic uveitis, incidence stratified by sex and race and ethnicity, and median charge per syphilitic uveitis hospital admission. Results: From the Nationwide Inpatient Sample, inpatient data from 444 674 patients (median [IQR] age, 53 [37-67] years; 54.8% male) were analyzed. There were an estimated 5581 syphilitic uveitis-related hospitalizations during the 10-year study period. The median (IQR) age of individuals with syphilitic uveitis was 45 (35-55) years, and 4395 patients (78.9%) were male. Syphilitic uveitis disproportionately affected African American individuals (1787 patients [32%], although they compose 13.6% of the population) and those belonging to the lowest median household income quartile (2163 [38.8%]). The national incidence was 0.15 per 100 000 population and showed an increasing trend over the years, with the lowest incidence in 2011 (0.08 per 100 000 population) and the highest incidence in 2019 (0.23 per 100 000 population; P = .04). Regional analysis showed an increase in incidence across all 4 US geographical regions. A total of 1293 patients (23.2%) had comorbid AIDS. Conclusion and Relevance: Although this cross-sectional study only captured inpatient diagnosis, an increasing incidence of syphilitic uveitis-related hospitalizations was observed in the US between 2010 and 2019. Given the concomitant national shortage of injectable penicillin G, results suggest that clinicians should maintain a high index of suspicion for syphilis when evaluating patients with intraocular inflammation.

[Clinical and etiological characteristics of infectious vulvovaginitis in children in Zhejiang province from 2009 to 2019].

Objective: To explore the clinical characteristics, common pathogens in children with vulvovaginitis. Methods: This was a retrospective cases study. A total of 3 268 children with vulvovaginitis were enrolled, who visited the Department of Pediatric and Adolescent Gynecology, Children's Hospital, Zhejiang University School of Medicine from January 2009 to December 2019. Patients were divided into 3 groups according to the age of <7, 7-<10 and 10-18 years. Patients were also divided in to 4 groups according to the season of first visit. The pathogen distribution characteristics of infective vulvovaginitis were compared between the groups. Their clinical data were collected and then analyzed by χ2 test. Results: The were 3 268 girls aged (6.2±2.5) years. There were 1 728 cases (52.9%) aged <7 years, 875 cases (26.8%) aged 7-<10 years, and 665 cases (20.3%) aged 10-18 years. Of these cases, 2 253 cases (68.9%) were bacterial vulvovaginitis, 715 cases (21.9%) were fungal vulvovaginitis and 300 cases (9.2%) were vulvovaginitis infected with other pathogens. Bacterial culture of vaginal secretions was performed in 2 287 cases, and 2 287 strains (70.0%) of pathogens were detected, of which the top 5 pathogens were Streptococcus pyogenes (745 strains, 32.6%), Haemophilus influenzae (717 strains, 31.4%), Escherichia coli (292 strains, 12.8%), Staphylococcus aureus (222 strains, 9.7%) and Klebsiella pneumoniae (67 strains, 2.9%). Regarding different age groups, H.influenzae was the most common in children under 7 years of age (40.3%, 509/1 263), S.pyogenes (41.9%, 356/849) was predominantly in children aged 7 to 10 years, and E.coli was predominant in children aged 10 to 18 years (26.3%, 46/175). Susceptibility results showed that S.pyogenes was susceptible to penicillin G (610/610, 100.0%), ceftriaxone (525/525, 100.0%), and vancomycin (610/610, 100.0%); the resistance rates to erythromycin and clindamycin were 91.9% (501/545)and 90.7% (495/546), respectively. For H.influenzae, 32.5% (161/496) produced ß-elactamase, and all strains were sensitive to meropenem (489/489, 100.0%) and levofloxacin (388/388, 100.0%), while 40.5% (202/499) were resistant to ampicillin. Among E.coli, all strains were sensitive to imipenem(100%, 175/175). The resistance rates of E.coli to levofloxacin and ceftriaxone were 29.1% (43/148) and 35.1% (59/168), respectively. A total of 48 strains of methicillin-resistant Staphylococcus aureus (MRSA) were isolated with a proportion of 28.3% (45/159) in 3 268 patients. The results of drug susceptibility test showed that all MRSA strains were sensitive to linezolid 100.0% (40/40), vancomycin (45/45, 100.0%), and tigecycline (36/36, 100.0%); the resistance rates of MRSA to penicillin G, erythromycin and clindamycin were 100% (45/45), 95.6% (43/45) and 88.9% (40/45), respectively. All methicillin-sensitive Staphylococcus aureus (MSSA) strains were sensitive to oxacillin (114/114, 100.0%), linezolid (94/94, 100.0%), vancomycin (114/114, 100.0%), and tigecycline (84/84, 100.0%); it's resistance rates to penicillin G, erythromycin and clindamycin were 78.1% (89/114), 59.7% (68/114) and 46.5% (53/114), respectively. The drug resistance rate of MSSA to penicillin G, erythromycin and clindamycin were lower than those of MRSA (χ²=11.71,19.74,23.95, respectively, all P<0.001). Conclusions: The age of consultation for pediatric infectious vulvovaginitis is mainly around 6 years. The most common pathogens are S.pyogenes, H.influenzae and Escherichia coli. Third generation cephalosporins can be used as the first choice of empirical anti-infection drugs. However, the results of drug susceptibility should be considered for targeted treatment.

Antimicrobial Resistance and Molecular Typing of Neisseria gonorrhoeae Isolates From the Eastern Cape Province in South Africa.

BACKGROUND: There is a paucity of Neisseria gonorrhoeae antimicrobial resistance data from resource-constrained settings because of the lack of diagnostic testing and limited scale of surveillance programs. This study aimed to determine the antimicrobial resistance profile of N. gonorrhoeae in the rural Eastern Cape province of South Africa. METHODS: Specimens for N. gonorrhoeae culture were obtained from men with urethral discharge and women with vaginal discharge attending primary health care facilities. Direct inoculation of the agar plates was performed followed by culture and drug susceptibility testing using the Etest at the laboratory. Whole-genome sequencing of the isolates was performed to identify resistance-determining variants. RESULTS: One hundred N. gonorrhoeae isolates were obtained. Most strains were nonsusceptible to ciprofloxacin (76%), tetracycline (75%), and penicillin G (72%). The gyrA S91F mutation was present in 68 of 72 ciprofloxacin-resistant isolates (94%), with concurrent parC mutations in 47 of 68 (69%); gyrA I250M was the only mutation in 4 other resistant strains. One azithromycin-resistant isolate was identified with a minimal inhibitory concentration (MIC) of 8.0 mg/L and the 23S rDNA gene mutation C2597T. The median MIC of cefixime was 0.016 mg/L (range, 0.016-0.064 mg/L), and that of ceftriaxone was 0.016 mg/L (range, 0.016 mg/L). Whole-genome sequencing showed 58 sequence types as revealed in N. gonorrhoeae sequence typing for antimicrobial resistance and 70 sequence types in N. gonorrhoeae multiantigen sequence typing. CONCLUSIONS: This study confirmed high rates of N. gonorrhoeae antimicrobial resistance to ciprofloxacin, penicillin G, and tetracycline in our setting. The MICs of cephalosporins are reassuring for ceftriaxone use in syndromic treatment regimens, but the identification of azithromycin resistance warrants further attention.

Rapid Environmental Impact Assessment of Penicillin G in a Veterinary Product Using an Original Software Method and Monitoring by SPE-Online-UHPLC-MS/MS.

Veterinary antibiotics have become a major concern due to potential environmental effects. This study presents an investigation of the exposure and environmental effects of the veterinary medicinal product in powder form, administered in the drinking water of piglets, chickens and turkeys, containing 250 mg/g penicillin G (benzylpenicillin potassium), performed according to the European Medicines Agency (EMEA) guideline and the results obtained by an analytical method based on online solid-phase extraction, ultra-high performance liquid chromatography coupled to a triple quadrupole mass spectrometer (SPE-online-UHPLC-MS/MS). The study presents the determination of the environmental risk and through an original, interactive, fast software method, created on the basis of a proprietary calculation algorithm that goes through all the prescriptions and recommendations of the EMEA guide. The results demonstrated that the concentration value for penicillin G determined in surface water by SPE-online-UHPLC-MS/MS is much lower than that predicted by calculation (predictable concentration in surface water, PECsurface water = 37.66 µg/L and the concentration SPE-online-UHPLC-MS/MS = 0.032 µg/L). Both results lead to a sub-unit risk quotient (R) indicating that the treatment carried out with the considered veterinary product does not present any risk to the environment.

Biofilm formation on endovascular aneurysm repair (EVAR) grafts-a proof of concept in vitro model.

OBJECTIVES: An endovascular aneurysm repair (EVAR) graft is a catheter-implanted vascular prosthesis and is the preferred treatment for patients with aortic aneurysm. If an EVAR graft becomes the focus of infection, the treatment possibilities are limited because it is technically difficult to remove the graft to obtain source control. This study examines whether Pseudomonas aeruginosa and Staphylococcus aureus form biofilm on EVAR prostheses. METHODS: EVAR graft sections were exposed to bacteria at 102 or 108 colony forming units (CFU)/mL in lysogeny broth and Krebs-Ringer at 37°C, bacterial biofilm formation was evaluated by scanning electron microscopy and counting CFU on the graft sections after antibiotic exposure at × 10 minimal inhibitory concentration. Bacteria were tested for tolerance to benzylpenicillin, tobramycin, and ciprofloxacin. RESULTS: Bacterial exposure for 15 minutes established biofilms on all prosthesis fragments (6/6 replicates). After 4 hours, bacteria were firmly attached to the EVAR prostheses and resisted washing. After 18-24 hours, the median CFU/g of EVAR graft reached 5.2 × 108 (1.15 × 108-1.1 × 109) for S. aureus and 9.1 × 107 (3.5 × 107-6.25 × 108) for P. aeruginosa. Scanning electron microscopy showed bacterial attachment to the graft pieces. There was a time-dependent development of tolerance with approximately 20 (tobramycin), 560 (benzylpenicillin), and 600 (ciprofloxacin) times more S. aureus surviving antibiotic exposure in 24- compared with 0-hour-old biofilm. Five (tobramycin) and 170 times (ciprofloxacin) more P. aeruginosa survived antibiotic exposure in 24- compared with 0-hour-old biofilms. DISCUSSION: Our results show that bacteria can rapidly adhere to and subsequently form antibiotic-tolerant biofilms on EVAR graft material in concentrations equivalent to levels seen in transient bacteraemia in vivo. Potentially, the system can be used for identifying optimal treatment combinations for infected EVAR prosthesis.

Case Report: Multimode Imaging of Chronic Syphilitic Chorioretinitis.

SIGNIFICANCE: The clinical manifestations of ocular syphilis may mimic those of other diseases, which may result in a missed diagnosis and delayed treatment. PURPOSE: We describe multimodal imaging findings and treatment outcomes of a patient with chronic syphilitic chorioretinitis. CASE REPORT: A 40-year-old male patient complained of progressive decreased visual acuity of his left eye for more than 1 year. The best-corrected visual acuity was 20/20 in the right eye and 3/50 in the left eye. Relative afferent pupillary defect and 1+ vitreous cells were detected in the left eye. The authors performed fundus examination, fluorescence angiography, ultrawide-field fundus autofluorescence, structure optical coherence tomography, wide-field montage optical coherence tomography angiography, and visual field. Laboratory tests including a toluidine red unheated serum test (1:32) and the Treponema pallidum antibody (9.01S/CO) showed positive results. Chronic syphilitic chorioretinitis was diagnosed in both eyes. The patient was admitted for administration of intravenous penicillin G for 14 days, followed by intramuscular benzathine penicillin G weekly for three doses. Six months after treatment, the toluidine red unheated serum test ratio had decreased to 1:2 (positive). The best-corrected visual acuity was 20/20 in the right eye and 6/20 in the left eye. The reexamination results showed that the ocular structure and capillaris flow partially recovered. CONCLUSIONS: Chronic syphilitic chorioretinitis profoundly affects the structure of the retina and choroid; however, eyes may partially recover after an effective treatment. Ultrawide-field imaging technology has several advantages, such as broader imaging field and more details provided, in determining syphilis-induced ocular disorders.

Treatment outcomes with benzylpenicillin and non-benzylpenicillin antibiotics, and the performance of the penicillin zone-edge test versus molecular detection of blaZ in penicillin-susceptible Staphylococcus aureus (PSSA) bacteraemia.

OBJECTIVES: The blaZ gene encodes penicillinase, which inactivates penicillin. As there were reports on suboptimal sensitivity for the penicillin zone-edge test, a phenotypic method for blaZ detection, we investigated treatment outcomes in patients with penicillin-susceptible Staphylococcus aureus (PSSA) bacteraemia (phenotypically negative for penicillinase), subjecting isolates to molecular testing for blaZ retrospectively. PATIENTS AND METHODS: A retrospective cohort study was conducted on 121 patients with a first episode of PSSA bacteraemia from 1 January 2012 to 31 October 2015 at Tan Tock Seng Hospital (TTSH), Singapore. Patients were grouped into IV benzylpenicillin and non-benzylpenicillin groups. The primary outcome was overall treatment failure, defined as either 30 day all-cause mortality and/or 90 day relapse. The penicillin (P10) zone-edge test was repeated on archived PSSA isolates, concurrently with penicillin MIC determination via gradient diffusion and PCR for blaZ. RESULTS: Among 121 patients, 57 patients (47.1%) received IV benzylpenicillin as the predominant antibiotic. There was no significant difference in overall treatment failure between treatment with the benzylpenicillin [7/57 (12.3%)] versus non-benzylpenicillin groups [12/64 (18.8%)] (P = 0.33) or cloxacillin/cefazolin [6/37 (16.2%)] (P = 0.59). For 112 PSSA isolates available for testing, repeat penicillin zone-edge testing was negative for penicillinase production, corroborating previous results. A single PSSA isolate with a negative penicillin zone-edge test was found to be positive for blaZ. CONCLUSIONS: We found no differences in overall treatment failure between patients with PSSA bacteraemia treated with benzylpenicillin, anti-staphylococcal ß-lactams cefazolin/cloxacillin and other antimicrobials, when using the penicillin zone-edge test as the phenotypic method for blaZ screening.

Tratamento da sífilis congênita e sua repercussão na rotina neonatal / Treatment of congenital syphilis and its impact on the neonatal routine

Introdução: A sífilis é uma doença infecto-contagiosa transmitida pela via sexual (sífilis adquirida) e verticalmente (sífilis congênita) durante a gestação pela placenta da mãe para o feto. Objetivos: Apresentar o tratamento proposto pelo Ministério da Saúde através do protocolo clínico atual para Sífilis transmitida verticalmente e relatar se foi uma opção viável, já que não havia a disponibilidade da Benzilpenicilina Procaína em todos os estados brasileiros; analisar quantas internações por um período mínimo de 10 dias poderiam ter sido evitadas se houvesse a disponibilidade da Benzilpenicilina Procaína conforme está no protocolo do Ministério da Saúde sendo notada a falta de tal medicação, prioritariamente, no Estado de São Paulo. Método: Estudo retrospectivo, investigativo, com revisão de prontuários de 23 gestantes com Sífilis e os respectivos recém-nascidos durante todo o ano de 2020, na maternidade de um hospital escola em Catanduva-SP. Resultados: Todos os recém-nascidos de mães com Sífilis na gestação recebiam o tratamento e/ou o seguimento para Sífilis Congênita de acordo com o teste - VDRL colhido ao nascimento. O tratamento para Sífilis Congênita sem Neurossífilis foi realizado com Benzilpenicilina Cristalina durante 10 dias, por via endovenosa, em quatorze bebês (56,5%). Destas 14 internações, apenas um caso de Sífilis Congênita com Neurossífilis realmente necessitava permanecer internada para tratamento endovenoso com Benzilpenicilina Cristalina. Uma mãe evadiu-se do hospital com o recém-nascido e não completou o tratamento no hospital-escola deste estudo, e para oito (34,8%) bebês definiu-se apenas o seguimento ambulatorial como conduta de acordo com as diretrizes do Ministério da Saúde. Em relação às intercorrências acometidas aos bebês que ficaram internados foram basicamente a dificuldade de amamentação e a perda do acesso venoso periférico com recorrência. Conclusão: A discordância nas Diretrizes do Ministério da Saúde para o tratamento da Sífilis Congênita, mesmo as mais atualizadas (2020), ocorre em decorrência da indisponibilidade desta medicação de baixo custo, Benzilpenicilina Procaína, aplicada intramuscular, o que facilita sua administração a nível de Unidades Básicas de Saúde e dispensa a internação hospitalar de bebês com necessidade de tratamento para a Sífilis Congênita sem Neurossífilis. Com isso, o trabalho sugere que a disponibilização da Benzilpenicilina Procaína para o tratamento dos casos simples de Sífilis Congênita culminará em incontáveis benefícios para o binômio mãe-bebê

Introduction: Syphilis is an infectious contagious disease transmitted sexually (acquired syphilis) and vertically (congenital syphilis) during pregnancy through the placenta from the mother to the fetus. Objectives: To prove whether the treatment proposed by the Ministry of Health through the current clinical protocol for vertically transmitted syphilis is a viable option, since Benzylpenicillin Procaine is not available in all Brazilian states; and to analyze how many hospitalizations for a minimum period of 10 days could have been avoided if Benzylpenicillin Procaine had been available as per the Ministry of Health protocol, with the lack of such medication being noted, primarily, in the State of São Paulo. Method: Retrospective, investigative study, reviewing the medical records of 23 pregnant women with Syphilis and their newborns throughout the year 2020, in the maternity hospital of a teaching hospital in Catanduva-SP. Results: All newborns of mothers with Syphilis during pregnancy received treatment and/or follow-up for Congenital Syphilis according to the test - VDRL collected at birth. Treatment for Congenital Syphilis without Neurosyphilis was performed with crystalline Benzylpenicillin for 10 days, intravenously, in fourteen babies (56.5%). Of these 14 hospitalizations, only one case of Congenital Syphilis with Neurosyphilis really needed to remain hospitalized for intravenous treatment with Crystalline Benzylpenicillin. One mother escaped from the hospital with the newborn and did not complete the treatment at the Hospital-Escola, and eight (34.8%) babies were defined only as outpatient follow-up as conduct in accordance with the guidelines of the Ministry of Health. Regarding the complications affected by the babies who were hospitalized, they were basically the difficulty in breastfeeding and the loss of peripheral venous access with recurrence. Conclusion: Disagreement in the Guidelines of the Ministry of Health for the treatment of Congenital Syphilis, even the most up-to-date ones (2020) occurs due to the unavailability of this low-cost medication, Benzylpenicillin Procaine, applied intramuscularly, which facilitates its administration at the Unit level Health Basics and waives the hospitalization of babies in need of treatment for Congenital Syphilis without Neurosyphilis. With this, the work suggests that the availability of Procaine Benzylpenicillin for the treatment of simple cases of Congenital Syphilis will culminate in countless benefits for the mother baby binomial

Introducción: La sífilis es una enfermedad infectocontagiosa de transmisión sexual (sífilis adquirida) y vertical (sífilis congénita) durante el embarazo a través de la placenta de la madre al feto. Objetivos: Probar si el tratamiento propuesto por el Ministerio de Salud a través del protocolo clínico vigente para la sífilis de transmisión vertical es una opción viable, ya que la Bencililpenicilina Procaína no está disponible en todos los estados brasileños; y analizar cuántas hospitalizaciones por un período mínimo de 10 días se podrían haber evitado si la bencililpenicilina procaína hubiera estado disponible según el protocolo del Ministerio de Salud, observándose la falta de dicha medicación, principalmente, en el estado de São Paulo. Método: Estudio retrospectivo, investigativo, revisando las historias clínicas de 23 gestantes con Sífilis y sus recién nacidos a lo largo del año 2020, en la maternidad de un hospital universitario de Catanduva-SP. Resultados: Todos los recién nacidos de madres con Sífilis durante el embarazo recibieron tratamiento y/o seguimiento para Sífilis Congénita según prueba - VDRL recogido al nacer. El tratamiento de la Sífilis Congénita sin Neurosífilis se realizó con Bencilpenicilina cristalina durante 10 días, por vía intravenosa, en catorce bebés (56,5%). De estas 14 hospitalizaciones, solo un caso de Sífilis Congénita con Neurosífilis necesitó realmente permanecer hospitalizado para recibir tratamiento intravenoso con Bencilpenicilina Cristalina. Una madre se escapó del hospital con el recién nacido y no completó el tratamiento en el hospital escuela en este estudio, y ocho (34,8%) bebés fueron definidos solo como seguimiento ambulatorio como conducta de acuerdo con las directrices del Ministerio de Salud. En cuanto a las complicaciones que afectaron a los bebés que fueron hospitalizados fueron básicamente la dificultad en la lactancia y la pérdida del acceso venoso periférico con recurrencia. Conclusión: La inconformidad en las Directrices del Ministerio de Salud para el tratamiento de la Sífilis Congénita, incluso las más actualizadas (2020) se da por la indisponibilidad de este medicamento de bajo costo, la Bencilpenicilina Procaína, de aplicación intramuscular, que facilita su administración a nivel de Unidad Básicos de Salud y renuncia a la hospitalización de bebés con necesidad de tratamiento por Sífilis Congénita sin Neurosífilis. Con ello, el trabajo sugiere que la disponibilidad de la Bencilpenicilina Procaína para el tratamiento de casos simples de Sífilis Congénita culminará en innumerables beneficios para el binomio madre-bebé

Neurosyphilis in a young man having HIV infection, blurring of vision and skin rash.

We report a 22-year-old homosexual man, a known case of HIV-1 infection but non-compliant to medications, who presented with a 5-month history of progressive painless non-pruritic coin-shaped skin lesions and recent gradual bilateral blurring of vision. His history divulged a primary syphilitic event 1 year prior to the present clinical manifestations. Investigation led to the diagnosis of neurosyphilis with ocular involvement with concurrent signs of secondary syphilis. Treatment with aqueous crystalline penicillin G, ophthalmic steroid and tropicamide drops, and topical emollients resulted in significant clinical improvement of ocular symptoms and skin lesions. The diagnosis of neurosyphilis requires a high degree of clinical suspicion and should be included in the differential diagnosis of unexplained ocular symptoms, particularly in men who have sex with men and HIV-infected patients. This is necessary for the early diagnosis, appropriate management, and good outcome of these patients.

Selection and Molecular Response of AHL-lactonase (aiiA) Producing Bacillus sp. Under Penicillin G-induced Conditions.

Quorum sensing (QS) is the process by which microorganisms employ chemicals called autoinducers (AIs) to communicate with their population. The QS mechanism generally controls the expression of the virulence related genes in bacteria. N-acyl homoserine lactones (AHLs) are the most widespread QS molecules. Due to their diverse AHL-lactonase activities, Bacillus species make particularly suitable candidates for procedures such as demolition of pathogenic bacterial QS signals and bioremediation of ß-lactam antibiotics from contaminated environments. In this study, seven Bacillus strains with Quorum quenching (QQ) activity were isolated using an enrichment medium supplemented with Penicillin G (PenG). The AHL-lactonase encoding gene (aiiA) was amplified by PCR and sequenced. Amino acid sequences underwent multiple sequence alignment. Docking studies were carried out with both C6HSL and PenG ligand using AutoDock tools. The aiiA amino acid sequences of the isolates were found to be well conserved. Furthermore, amino acid sequence alignment revealed that 74.9% of amino acid sequences were conserved in the genus Bacillus. Docking of the C6HSL to wild type (3DHA) and H97D variant reduced the docking score by only 0.1 kcal/mol for the mutated protein. When PenG docked with a higher (1.5 kcal/mol) score as a ligand to wild-type and mutant receptors, the docking score for the mutated protein likewise decreased by 0.1 kcal/mol. This research contributed to the diversification of organisms with QQ activity and beta-lactam antibiotic resistance. It also clarified the binding score of the PenG ligand to the Bacillus AHL lactonase molecule for the first time.