Antibiotics

13 pages, 240 KiB

Open AccessArticle

Development and Pilot of an Interactive Online Course on Antimicrobial Stewardship in Companion Animals

by Nonke E. M. Hopman, Jaap A. Wagenaar, Ingeborg M. van Geijlswijk and Els M. Broens

Antibiotics 2021, 10(5), 610; https://doi.org/10.3390/antibiotics10050610 - 20 May 2021

Cited by 1 | Viewed by 3053

A holistic approach to antimicrobial use (AMU) and prescribing is needed to combat the problem of antimicrobial resistance (AMR). Previously, an antimicrobial stewardship programme (ASP) was developed, introduced, and evaluated in 44 Dutch companion animal clinics, which resulted in an optimization of AMU. [...] Read more.

A holistic approach to antimicrobial use (AMU) and prescribing is needed to combat the problem of antimicrobial resistance (AMR). Previously, an antimicrobial stewardship programme (ASP) was developed, introduced, and evaluated in 44 Dutch companion animal clinics, which resulted in an optimization of AMU. As a follow-up to this, an online course was developed to promote awareness of AMU, AMR, and responsible antimicrobial prescribing. The aim of this paper is to describe the development and pilot, including evaluation, of this course, which will be disseminated more widely among Dutch companion animal veterinarians. The interactive programme consists of a major e-learning component and two online, face-to-face meetings. The course comprises five different parts corresponding with five consecutive weeks. Theory on several topics is offered, for example on AMU and AMR in general, Dutch regulations and guidelines on veterinary AMU, behavioural change, and possible methods to quantify AMU. Additionally, several assignments are offered, for example to reflect upon one’s own current antimicrobial prescribing behaviour. Interactive discussion and peer-to-peer learning are promoted. Since September 2020, the course has been offered in a pilot phase, and the feedback is promising. Evaluation of the pilot phase will result in recommendations for further optimization and dissemination. Full article

(This article belongs to the Special Issue Antimicrobial Stewardship in Veterinary Medicine)

8 pages, 1292 KiB

Open AccessBrief Report

Inhibition of LpxC Increases the Activity of Iron Chelators and Gallium Nitrate in Multidrug-Resistant Acinetobacter baumannii

by Víctor Vinuesa, Raquel Cruces, Francesca Nonnoi and Michael J. McConnell

Antibiotics 2021, 10(5), 609; https://doi.org/10.3390/antibiotics10050609 - 20 May 2021

Cited by 5 | Viewed by 2509

Abstract

Infections caused by multidrug-resistant Acinetobacter baumannii would benefit from the development of novel treatment approaches. Compounds that interfere with bacterial iron metabolism, such as iron chelators and gallium nitrate, have previously been shown to have antimicrobial activity against A. baumannii. In this [...] Read more.

Infections caused by multidrug-resistant Acinetobacter baumannii would benefit from the development of novel treatment approaches. Compounds that interfere with bacterial iron metabolism, such as iron chelators and gallium nitrate, have previously been shown to have antimicrobial activity against A. baumannii. In this study, we characterize the effect of LpxC inhibitors on the antimicrobial activity of previously characterized iron chelators, 2,2′-bipyridyl (BIP) and deferiprone (DFP), and gallium nitrate (Ga(NO₃)₃) against A. baumannii reference strains and multidrug-resistant clinical isolates. The LpxC inhibitor LpxC-2 was synergistic with BIP for 30% of strains tested (FICI values: 0.38–1.02), whereas inhibition with LpxC-4 was synergistic with BIP for 60% of strains tested (FICI values: 0.09–0.75). In time–kill assays, combinations of BIP with both LpxC inhibitors demonstrated synergistic activity, with a more than 3 log₁₀ reduction in bacterial counts compared to BIP alone. LpxC-2 was synergistic with Ga(NO₃)₃ for 50% of strains tested (FICI values: 0.27–1.0), whereas LpxC-4 was synergistic with Ga(NO₃)₃ for all strains tested (FICI values: 0.08–≤0.50). In time–kill assays, combinations of Ga(NO₃)₃ with LpxC-2 and LpxC-4 decreased the growth of both strains compared to each compound separately; however, only the combination with LpxC-4 met the defined criteria for synergy. These results identify a novel synergy between two antimicrobial classes against A. baumannii strains. Full article

(This article belongs to the Section Novel Antimicrobial Agents)

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10 pages, 772 KiB

Open AccessArticle

Multi-Drug Resistant Organisms Infection Impact on Patients Length of Stay in Respiratory Care Ward

by Yi-Ping Chen, Xian-Wen Tasi, Ko Chang, Xuan-Di Cao, Jung-Ren Chen and Chien-Sen Liao

Antibiotics 2021, 10(5), 608; https://doi.org/10.3390/antibiotics10050608 - 20 May 2021

Cited by 5 | Viewed by 3182

Abstract

This study aimed to investigate the effects of multi-drug-resistant organism (MDRO) infection and other factors on the length of hospital stay (LOS) of patients in the respiratory care ward (RCW) of a regional hospital in Taiwan. In this retrospective study, we collected cases [...] Read more.

This study aimed to investigate the effects of multi-drug-resistant organism (MDRO) infection and other factors on the length of hospital stay (LOS) of patients in the respiratory care ward (RCW) of a regional hospital in Taiwan. In this retrospective study, we collected cases from MDRO-infected patients in the RCW from January 2016 to March 2020. The RCW comprises 13 beds in total. There were 106 infected patients, of which 42 were in the case group (infected with MDROs) and 64 were in the control group (not infected with MDROs). Clinical specimens were inoculated in a selective medium to isolate the pathogenic bacteria by standard procedures. The results showed the main factors affecting the LOS were: patients with MDRO infection, patients discharged from the RCW, and patients who underwent catheterization. The LOS of patients infected with MDROs was significantly longer than that of patients without MDRO infection (β = 0.55, 95% CI = 0.02–1.09), with the case group and the control group being 479.8 ± 546.5 and 307.3 ± 436.2 days, respectively. Infection with carbapenem-resistant Pseudomonas aeruginosa (CRPA) was associated with a longer LOS than other MDRO strains. These findings have important implications for infection control in RCW and in better tracking the health of patients. Full article

(This article belongs to the Special Issue Multi-Drug Resistant Gram-Negative Microorganisms: Epidemiology, Treatment and Alternative Approach)

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13 pages, 498 KiB

Open AccessReview

Sensitivity of Staphylococcal Biofilm to Selected Compounds of Plant Origin

by Denis Swolana, Małgorzata Kępa, Agata Kabała-Dzik, Radosław Dzik and Robert D. Wojtyczka

Antibiotics 2021, 10(5), 607; https://doi.org/10.3390/antibiotics10050607 - 20 May 2021

Cited by 9 | Viewed by 3091

Abstract

Staphylococcus epidermidis is a bacterium that belongs to the human microbiota. It is most plentiful on the skin, in the respiratory system, and in the human digestive tract. Moreover, it is the most frequently isolated microorganism belonging to the group of Coagulase Negative [...] Read more.

Staphylococcus epidermidis is a bacterium that belongs to the human microbiota. It is most plentiful on the skin, in the respiratory system, and in the human digestive tract. Moreover, it is the most frequently isolated microorganism belonging to the group of Coagulase Negative Staphylococci (CoNS). In recent years, it has been recognized as an important etiological factor of mainly nosocomial infections and infections related to the cardiovascular system. On the other hand, Staphylococcus aureus, responsible for in-hospital and out-of-hospital infections, is posing an increasing problem for clinicians due to its growing resistance to antibiotics. Biofilm produced by both of these staphylococcal species in the course of infection significantly impedes therapy. The ability to produce biofilm hinders the activity of chemotherapeutic agents—the only currently available antimicrobial therapy. This also causes the observed significant increase in bacterial resistance. For this reason, we are constantly looking for new substances that can neutralize microbial cells. In the present review, 58 substances of plant origin with antimicrobial activity against staphylococcal biofilm were replaced. Variable antimicrobial efficacy of the substances was demonstrated, depending on the age of the biofilm. An increase in the activity of the compounds occurred in proportion to increasing their concentration. Appropriate use of the potential of plant-derived compounds as an alternative to antibiotics may represent an important direction of change in the support of antimicrobial therapy. Full article

(This article belongs to the Special Issue Development of Antimicrobial Biomaterials and Natural Alternatives against Biofilms and Implant-Related Infections)

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15 pages, 5140 KiB

Open AccessArticle

Educational Video Improves Knowledge about Outpatients’ Usage of Antibiotics in Two Public Hospitals in Indonesia

by Fauna Herawati, Rika Yulia, Bustanul Arifin, Ikhwan Frasetyo, Setiasih, Herman J. Woerdenbag, Christina Avanti and Retnosari Andrajati

Antibiotics 2021, 10(5), 606; https://doi.org/10.3390/antibiotics10050606 - 20 May 2021

Cited by 4 | Viewed by 3971

Abstract

The inappropriate use or misuse of antibiotics, particularly by outpatients, increases antibiotic resistance. A lack of public knowledge about “Responsible use of antibiotics” and “How to obtain antibiotics” is a major cause of this. This study aimed to assess the effectiveness of an [...] Read more.

The inappropriate use or misuse of antibiotics, particularly by outpatients, increases antibiotic resistance. A lack of public knowledge about “Responsible use of antibiotics” and “How to obtain antibiotics” is a major cause of this. This study aimed to assess the effectiveness of an educational video about antibiotics and antibiotic use to increase outpatients’ knowledge shown in two public hospitals in East Java, Indonesia. A quasi-experimental research setting was used with a one-group pre-test—post-test design, carried out from November 2018 to January 2019. The study population consisted of outpatients to whom antibiotics were prescribed. Participants were selected using a purposive sampling technique; 98 outpatients at MZ General Hospital in the S regency and 96 at SG General Hospital in the L regency were included. A questionnaire was used to measure the respondents’ knowledge, and consisted of five domains, i.e., the definition of infections and antibiotics, obtaining the antibiotics, directions for use, storage instructions, and antibiotic resistance. The knowledge test score was the total score of the Guttman scale (a dichotomous “yes” or “no” answer). To determine the significance of the difference in knowledge before and after providing the educational video and in the knowledge score between hospitals, the (paired) Student’s t-test was applied. The educational videos significantly improved outpatients’ knowledge, which increased by 41% in MZ General Hospital, and by 42% in SG General Hospital. It was concluded that an educational video provides a useful method to improve the knowledge of the outpatients regarding antibiotics. Full article

(This article belongs to the Special Issue Antimicrobial Use, Resistance and Stewardship)

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20 pages, 1491 KiB

Open AccessArticle

The Perfect Condition for the Rising of Superbugs: Person-to-Person Contact and Antibiotic Use Are the Key Factors Responsible for the Positive Correlation between Antibiotic Resistance Gene Diversity and Virulence Gene Diversity in Human Metagenomes

by Célia P. F. Domingues, João S. Rebelo, Joël Pothier, Francisca Monteiro, Teresa Nogueira and Francisco Dionisio

Antibiotics 2021, 10(5), 605; https://doi.org/10.3390/antibiotics10050605 - 20 May 2021

Cited by 10 | Viewed by 4359

Abstract

Human metagenomes with a high diversity of virulence genes tend to have a high diversity of antibiotic-resistance genes and vice-versa. To understand this positive correlation, we simulated the transfer of these genes and bacterial pathogens in a community of interacting people that take [...] Read more.

Human metagenomes with a high diversity of virulence genes tend to have a high diversity of antibiotic-resistance genes and vice-versa. To understand this positive correlation, we simulated the transfer of these genes and bacterial pathogens in a community of interacting people that take antibiotics when infected by pathogens. Simulations show that people with higher diversity of virulence and resistance genes took antibiotics long ago, not recently. On the other extreme, we find people with low diversity of both gene types because they took antibiotics recently—while antibiotics select specific resistance genes, they also decrease gene diversity by eliminating bacteria. In general, the diversity of virulence and resistance genes becomes positively correlated whenever the transmission probability between people is higher than the probability of losing resistance genes. The positive correlation holds even under changes of several variables, such as the relative or total diversity of virulence and resistance genes, the contamination probability between individuals, the loss rate of resistance genes, or the social network type. Because the loss rate of resistance genes may be shallow, we conclude that the transmission between people and antibiotic usage are the leading causes for the positive correlation between virulence and antibiotic-resistance genes. Full article

(This article belongs to the Special Issue Antibiotic Resistance Genes: Spread and Evolution)

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Each human being in a community is colonized by bacteria (as an integral part of the microbiome). Some bacteria contain antibiotic resistance genes and/or virulence genes. These bacteria can be transmitted from one person to another, which includes the passage of resistance and/or virulence genes that they may contain. All the individuals in the community are organized in a network of contacts. In the regular network, each individual is linked to other four individuals (the four closest individuals; n = 4). The other two types of network were constructed following Watts and Strogatz method [<a href="#B37-antibiotics-10-00605" class="html-bibr">37</a>]. From the regular network, each connection has a probability p to change, that is, each individual can be reconnected to another one in the network. Therefore, the value of p defines the type of network: (i) p = 0 the network remains regular; (ii) p = 0.5 is a small-world network; (iii) p = 1 the network becomes random. Full article ">Figure 2
Flowchart of the program. After the network’s construction, the program performs several cycles where, eventually, there is gene transfer between nodes (people). Some individuals get sick and take antibiotics. Some genes are lost due to antibiotic pressure or the fitness cost imposed by resistance genes. Full article ">Figure 3
Effect of the gene transmission probability. (A–F): the relationship between the diversity of resistance genes (vertical axes) and the diversity of virulence genes (horizontal axes). Each dot represents the case of an individual metagenome. (A,B): disappearance of the diversity of virulence genes; (C,D): positive correlation between the diversity of resistance genes and the diversity of virulence genes; (E,F): positive correlation between the diversity of resistance genes and the diversity of virulence, with many individuals having a high diversity of the two gene types. Parameters as follows. In all cases, we set resistance genes loss rate = 0. In (A), when the gene transmission probability is low (0.0005), virulence genes disappeared from the network. In (B), gene transmission probability = 0.0025 (R = 0.309, slope = 11.00, p-value = 1.47 × 10−23). In (C), gene transmission probability = 0.005 (R = 0.934, slope = 0.798, p-value = ~0). In (D), gene transmission probability = 0.01 (R = 0.973, slope = 0.757, p-value = ~0). In (E), gene transmission probability = 0.015 (R = 0.972, slope = 0.754, p-value = ~0). In (F), gene transmission probability = 0.02 (R = 0.976, slope = 0.751, p-value = ~0). Full article ">Figure 4
Effect of the relative values of the gene transmission probability and the resistance genes loss rate. (A,B): the relationship between the diversity of virulence genes (horizontal axes) and the diversity of resistance genes (vertical axes). Each dot represents the case of an individual metagenome. In both (A) and (B), the gene transmission probability = 0.005. (A): resistance genes loss rate = 0, which is lower than the gene transmission probability, resulting in a positive slope; (R = 0.929, slope = 0.775, p-value ~ 0). (B): resistance genes loss rate = 0.03, which higher than the gene transmission probability, resulting in a negative slope; (R = -0.682, slope = −0.174, p-value = 1.19 × 10−137). (C): Slope of the regression between the diversity of virulence and resistance genes according to the loss rate (horizontal axes) and the gene transmission probability (vertical axes). Green: positive slopes; Red: negative slopes; Blue: the slope is not significantly different from zero (p-value ≥ 1 × 10−6). Full article ">

16 pages, 3024 KiB

Open AccessArticle

Enrofloxacin Dose Optimization for the Treatment of Colibacillosis in Broiler Chickens Using a Drinking Behaviour Pharmacokinetic Model

by Robin Temmerman, Ludovic Pelligand, Wim Schelstraete, Gunther Antonissen, An Garmyn and Mathias Devreese

Antibiotics 2021, 10(5), 604; https://doi.org/10.3390/antibiotics10050604 - 19 May 2021

Cited by 11 | Viewed by 5079

Abstract

Enrofloxacin is frequently administered via drinking water for the treatment of colibacillosis in broiler chickens. However, the EMA/CVMP has urged to re-evaluate historically approved doses, especially for antimicrobials administered via drinking water. In response, the objectives of this study were two-fold. First, to [...] Read more.

Enrofloxacin is frequently administered via drinking water for the treatment of colibacillosis in broiler chickens. However, the EMA/CVMP has urged to re-evaluate historically approved doses, especially for antimicrobials administered via drinking water. In response, the objectives of this study were two-fold. First, to evaluate the pharmacokinetics (PK) of enrofloxacin following IV, PO and drinking water administration. Second, to predict the efficacy of a range of doses in the drinking water for the treatment of APEC infections. For the first objective, PK parameters were estimated by fitting a one-compartmental model with a zero-order IV infusion and an oral absorption lag function to the simultaneously modelled IV and PO data. After fixing these parameter values, a drinking behaviour pharmacokinetic (DBPK) model was developed for the description and prediction of drinking water PK profiles by adding three model improvements (different diurnal and nocturnal drinking rates, inter-animal variability in water consumption and taking account of dose non-proportionality). The subsequent simulations and probability of target attainment (PTA) analysis predicted that a dose of 12.5 mg/kg/24 h is efficacious in treating colibacillosis with an MIC up to 0.125 μg/mL (ECOFF), whereas the currently registered dose (10 mg/kg/24 h) reaches a PTA of 66% at ECOFF. Full article

(This article belongs to the Special Issue Optimization and Improvement of Veterinary Antimicrobial Treatment to Reduce Antimicrobial Resistance)

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An overview of the PK modelling process. Using the data of the IV and PO single bolus administration (rich and sparsely sampled) animal trials, the parameters of the one-compartment model (Cl, V, Ka and F with an additional IV delay and Tlag) were estimated and subsequently fixed in further analyses. In order to model the drinking water administration modality and develop the DBPK model, three model improvements were implemented. First, diurnal and nocturnal infusion rates were chosen in order to mimic the drinking behaviour during the photo- and scoto-period, respectively. Second, the variability in drinking water behaviour and therefore in bioavailability was captured using an additional parameter, Drink. Third, accounting for the nonlinear relationship between dose and exposure, a covariate was introduced (dVddose) to decrease Drink and hence bioavailability proportionally with higher doses. Full article ">Figure 2
Stratified visual predictive check (VPC) of the different treatment groups ((a)—IV, (b)—PO richly sampled, (c)—PO sparsely sampled, (d)—drinking water administration 2.5 mg/kg, (e)—drinking water administration 5 mg/kg, (f)—drinking water administration 10 mg/kg, (g)—drinking water administration 15 mg/kg, and (h)—drinking water administration 20 mg/kg) with 200 replicates of each animal. Approximately 20% of the data should fall outside the plotted quantiles. Red lines: observed quantiles; black lines: predicted quantiles; blue circles: observed data. Full article ">Figure 3
Correlation between the observed water consumption and the estimated water uptake by the model for 15 animals. The observed water drunk over 24 h was determined by adding the estimated proportion of nocturnal drinking (10.5%) to the water uptake evaluated during the photoperiod. Coefficient of determination (R2) = 0.8020. Full article ">Figure 4
Probability of target attainment (PTA) for fAUC (48–72 h)/MIC ≥ 100 h in 75 simulated broiler chickens for four doses of enrofloxacin (10, 12.5, 20 and 50 mg/kg) administered via drinking water (five other simulated doses are omitted). The horizontal dotted line signifies the PTA of 90%. The relative frequency of the MIC distribution of APEC strains is taken from a recent study from Flanders (Belgium) [<a href="#B24-antibiotics-10-00604" class="html-bibr">24</a>]. Full article ">Figure 5
The green lines correspond with the time when enrofloxacin was introduced in the drinking water (9 a.m.) and kept for 24 h. This was done for three consecutive days. The red line indicates the cessation of enrofloxacin administration. The blue dotted lines relate to the six different sample points (7, 23, 31, 46, 55 and 79 h post administration). The grey areas indicate the periods that the light in the stable was turned off (scotoperiod, 11 p.m. to 7 a.m.), and the white areas correspond to the photoperiod. The black dotted line is an example of a PK profile of a representative broiler dosed at 10 mg/kg/24 h. Full article ">Figure 6
The correlation between AUC(0–72h) of enrofloxacin administered via the drinking water of 15 broiler chickens dosed at 20 mg/kg and the amount of water drunk over 16 h (the filming period) per bird the day before administration. Coefficient of determination (R2) = 0.8127. Full article ">

12 pages, 7471 KiB

Open AccessArticle

An Antisense yycF RNA Modulates Biofilm Organization of Methicillin-Resistant Staphylococcus aureus and Pathogenicity in a Rat Model of Osteomyelitis

by Shizhou Wu, Yunjie Liu, Lei Lei and Hui Zhang

Antibiotics 2021, 10(5), 603; https://doi.org/10.3390/antibiotics10050603 - 19 May 2021

Cited by 13 | Viewed by 2461

Abstract

Staphylococcus aureus (S. aureus) is one of most common opportunistic pathogens and is attributed to several human infections. The increasing incidence of methicillin-resistant S. aureus (MRSA) is a serious clinical threat for osteomyelitis crisis. The YycFG two-component system of S. aureus [...] Read more.

Staphylococcus aureus (S. aureus) is one of most common opportunistic pathogens and is attributed to several human infections. The increasing incidence of methicillin-resistant S. aureus (MRSA) is a serious clinical threat for osteomyelitis crisis. The YycFG two-component system of S. aureus regulates genes associated with biofilm formation. To investigate the potential role of an antisense yycF RNA in the regulation of transcription levels of yycF and associated effects on biofilm formation and pathogenicity, antisense yycF (ASyycF) RNA was detected by RT-PCR and 5′ RACE assays. ASyycF overexpression mutants were constructed, and the biofilm biomass was determined by crystal violet microtiter assay and scanning electron microscopy (SEM). Quantitative RT-PCR and Western blotting analyses were used to detect whether ASyycF overexpression inhibited the transcription and translation of biofilm-related genes. Then, a rat tibial infective model was used to evaluate the pathogenicity of ASyycF overexpression in vivo. ASyycF transcription led to reductions in YycF production and biofilm formation. Overexpression of ASyycF inhibited the transcription and translation of biofilm-related genes. The sensitivity to vancomycin was improved in ASyycF-overexpressing MRSA. Furthermore, ASyycF inhibited MRSA invasion in a rat tibial infection model. From this study, the expression of the YycF protein was found to be inversely correlated with different levels of ASyycF transcription. The biofilm biomass and pathogenicity decreased in the ASyycF-overexpressing mutant. Thus, the current evidence may support ASyycF as a supplementary strategy for managing S. aureus and MRSA infections. Full article

(This article belongs to the Special Issue Antibiotic Resistance and Treatment of MRSA Infection)

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Detection of antisense yycF RNA. (A) Total RNA samples were isolated from S. aureus to detect ASyycF RNA transcription by first cDNA strand synthesis and RT-PCR (red arrows). (B) Detection of the 5′ terminus of the ASyycF transcription by 5′ RACE (red arrows): (a) gene-specific outer primer; (b) gene-specific inner primer; (c) and (d) are 5′ RACE outer and inner primers, respectively. (C) Schematic of ASyycF showing that transcription starts within the 5′ terminus in the yycF open reading frame (ORF). The full length for ASyycF RNA is approximately 400 bp. Full article ">Figure 2
ASyycF modulated the bacterial growth and biofilm organization. (A) The growth curves for the Staphylococcus aureus. (B) Biomass was quantified by crystal violet staining. Optical densities at 600 nm were measured (n = 10, * p < 0.05). (C) SEM images of S. aureus ATCC29213, and methicillin-resistant Staphylococcus aureus (MRSA) strains after ASyycF overexpression. Full article ">Figure 3
Effect of biofilm formation on antibiotics sensitivity. (A) Intensity of fluorescence for S. aureus ATCC29213, and MRSA strains after ASyycF overexpression (scale bar = 100 μm). (B) Intensity of fluorescence comparisons and the intensities of S. aureus ATCC29213 were measured as reference (n = 10, * p < 0.05). (C) E-test for the sensitivity of MRSA to vancomycin. Full article ">Figure 4
ASyycF overexpression inhibited the transcription of biofilm-related genes. (A) Quantitative RT-PCR analysis showed the gene transcription in S. aureus, using 16S as an internal control (n = 10, * p < 0.05); (B) The productions of YycF were quantified in the cells of S. aureus for Western blotting (upper lane); the lower panel shows a Coomassie-stained gel supporting equal loading of each protein sample. (C) Quantitative analysis for the relative YycF protein amounts (n = 4, * p < 0.05). Full article ">Figure 5
ASyycF inhibited MRSA invasion in rat tibial infective model. (A) HE staining for histological evaluation (upper lane); the fluorescent labeled peptide nucleic acid in situ hybridization probing for S. aureus (lower lane). (B) Working model. Full article ">

8 pages, 1136 KiB

Open AccessCommunication

Pharmacodynamic Evaluation of a Single Dose versus a 24-Hour Course of Multiple Doses of Cefazolin for Surgical Prophylaxis

by Aaron Heffernan, Jowana Alawie, Steven C Wallis, Saiyuri Naicker, Santosh Adiraju, Jason A. Roberts and Fekade Bruck Sime

Antibiotics 2021, 10(5), 602; https://doi.org/10.3390/antibiotics10050602 - 19 May 2021

Cited by 2 | Viewed by 3387

Abstract

The optimal perioperative duration for the administration of cefazolin and other prophylactic antibiotics remains unclear. This study aimed to describe the pharmacodynamics of cefazolin for a single 2 g dose versus a 24 h course of a 2 g single dose plus a [...] Read more.

The optimal perioperative duration for the administration of cefazolin and other prophylactic antibiotics remains unclear. This study aimed to describe the pharmacodynamics of cefazolin for a single 2 g dose versus a 24 h course of a 2 g single dose plus a 1 g eight-hourly regimen against methicillin-susceptible Staphylococcus aureus. Static concentration time–kill assay and a dynamic in vitro hollow-fibre infection model simulating humanised plasma and interstitial fluid exposures of cefazolin were used to characterise the pharmacodynamics of prophylactic cefazolin regimens against methicillin-sensitive Staphylococcus aureus clinical isolates. The initial inoculum was 1 × 10⁵ CFU/mL to mimic a high skin flora inoculum. The static time–kill study showed that increasing the cefazolin concentration above 1 mg/L (the MIC) did not increase the rate or the extent of bacterial killing. In the dynamic hollow-fibre model, both dosing regimens achieved similar bacterial killing (~3-log CFU/mL within 24 h). A single 2 g dose may be adequate when low bacterial burdens (~10⁴ CFU/mL) are anticipated in an immunocompetent patient with normal pharmacokinetics. Full article

(This article belongs to the Special Issue Antibiotic Usage in Acute Situations)

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16 pages, 563 KiB

Open AccessArticle

Antibiotic Prescriptions among China Ambulatory Care Visits of Pregnant Women: A Nationwide Cross-Sectional Study

by Houyu Zhao, Mei Zhang, Jiaming Bian and Siyan Zhan

Antibiotics 2021, 10(5), 601; https://doi.org/10.3390/antibiotics10050601 - 19 May 2021

Cited by 14 | Viewed by 2805

Abstract

Background: Antibiotic use in pregnant women at the national level has rarely been reported in China. Objectives: We aimed to investigate antibiotic prescriptions during pregnancy in ambulatory care settings in China. Methods: Data of 4,574,961 ambulatory care visits of pregnant women from October [...] Read more.

Background: Antibiotic use in pregnant women at the national level has rarely been reported in China. Objectives: We aimed to investigate antibiotic prescriptions during pregnancy in ambulatory care settings in China. Methods: Data of 4,574,961 ambulatory care visits of pregnant women from October 2014 to April 2018 were analyzed. Percentages of Antibiotic prescriptions by different subgroups and various diagnosis categories and proportions of inappropriate antibiotic prescriptions for different subgroups were estimated. Food and Drug Administration (FDA) pregnancy categories were used to describe the antibiotic prescription patterns. The 95% confidence intervals (CIs) were estimated using the Clopper––Pearson method or Goodman method. Results: Among the 4,574,961 outpatient visits during pregnancy, 2.0% (92,514 visits; 95% CI, 2.0–2.0%) were prescribed at least one antibiotic. The percentage of antibiotic prescriptions for pregnant women aged >40 years was 4.9% (95% CI, 4.7–5.0%), whereas that for pregnant women aged 26–30 years was 1.5% (95% CI, 1.4–1.5%). In addition, percentages of antibiotic prescriptions varied among different trimesters of pregnancy, which were 5.4% (95% CI, 5.3–5.4%) for the visits in the first trimester of pregnancy and 0.5% (95% CI, 0.4–0.5%) in the third trimester of pregnancy. Furthermore, the percentages of antibiotic prescriptions substantially varied among different diagnosis categories and nearly three-quarters of antibiotic prescriptions had no clear indications and thus might be inappropriate. In total, 130,308 individual antibiotics were prescribed; among these, 60.4% (95% CI, 60.0–60.8%) belonged to FDA category B, 2.7% (95% CI, 2.1–3.5%) were classified as FDA category D and 16.8% (95% CI, 16.2–17.4%) were not assigned any FDA pregnancy category. Conclusions: Antibiotic prescriptions in ambulatory care during pregnancy were not highly prevalent in mainland China. However, a substantial proportion of antibiotics might have been prescribed without adequate indications. Antibiotics whose fetal safety has not been sufficiently illustrated were widely used in pregnant women. Full article

(This article belongs to the Special Issue Appropriateness of Antibiotics in China)

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17 pages, 2364 KiB

Open AccessArticle

Xanthones Active against Multidrug Resistance and Virulence Mechanisms of Bacteria

by Fernando Durães, Diana I. S. P. Resende, Andreia Palmeira, Nikoletta Szemerédi, Madalena M. M. Pinto, Gabriella Spengler and Emília Sousa

Antibiotics 2021, 10(5), 600; https://doi.org/10.3390/antibiotics10050600 - 19 May 2021

Cited by 25 | Viewed by 4860

Abstract

The emergence of multidrug and extensively drug-resistant pathogenic bacteria able to resist to the action of a wide range of antibiotics is becoming a growing problem for public health. The search for new compounds with the potential to help in the reversion of [...] Read more.

The emergence of multidrug and extensively drug-resistant pathogenic bacteria able to resist to the action of a wide range of antibiotics is becoming a growing problem for public health. The search for new compounds with the potential to help in the reversion of bacterial resistance plays an important role in current medicinal chemistry research. Under this scope, bacterial efflux pumps are responsible for the efflux of antimicrobials, and their inhibition could reverse resistance. In this study, the multidrug resistance reversing activity of a series of xanthones was investigated. Firstly, docking studies were performed in the AcrAB-TolC efflux pump and in a homology model of the NorA pump. Then, the effects of twenty xanthone derivatives on bacterial growth were evaluated in Staphylococcus aureus 272123 and in the acrA gene-inactivated mutant Salmonella enterica serovar Typhimurium SL1344 (SE03). Their efflux pump inhibitory properties were assessed using real-time fluorimetry. Assays concerning the activity of these compounds towards the inhibition of biofilm formation and quorum sensing have also been performed. Results showed that a halogenated phenylmethanamine xanthone derivative displayed an interesting profile, as far as efflux pump inhibition and biofilm formation were concerned. To the best of our knowledge, this is the first report of xanthones as potential efflux pump inhibitors. Full article

(This article belongs to the Special Issue The Role of Efflux Pump Inhibitor in Bacterial Multidrug Resistance)

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11 pages, 260 KiB

Open AccessArticle

Feline Otitis Externa Caused by Methicillin-Resistant Staphylococcus aureus with Mixed Hemolytic Phenotype and Overview of Possible Genetic Backgrounds

by Jana Avberšek, Bojan Papić, Darja Kušar, Vladimira Erjavec, Katja Seme, Majda Golob and Irena Zdovc

Antibiotics 2021, 10(5), 599; https://doi.org/10.3390/antibiotics10050599 - 18 May 2021

Cited by 3 | Viewed by 2589

Abstract

Methicillin-resistant Staphylococcus aureus (MRSA) is an important cause of nosocomial infections in humans, but its importance in small animal practice is increasing. Here, we present a case of feline otitis externa (OE) caused by MRSA; both hemolytic and nonhemolytic variants with a stable [...] Read more.

Methicillin-resistant Staphylococcus aureus (MRSA) is an important cause of nosocomial infections in humans, but its importance in small animal practice is increasing. Here, we present a case of feline otitis externa (OE) caused by MRSA; both hemolytic and nonhemolytic variants with a stable phenotype were recovered from the external auditory canal after infection was detected by routine otoscopy. One isolate per variant underwent antimicrobial susceptibility testing (AST) by broth microdilution method, conventional spa typing and whole-genome sequencing (WGS). The results showed that both variants were genetically related and were of sequence type (ST) 1327, SCCmec type IV and spa type t005. AST and WGS showed that both isolates were resistant to β-lactams and sensitive to all tested non-β-lactam antibiotics. Both isolates were pvl-negative, but encoded several other virulence genes (aur, hlgABC, sak, scn, seg, sei, sem, sen, seo and seu). Genetic background of the mixed hemolytic phenotype was not identified; no differences in the agr locus or other regulatory regions were detected. Three single-nucleotide polymorphisms were identified but could not be associated with hemolysis. This well-documented case of MRSA infection in companion animals adds to the reports of MRSA infections with a mixed hemolytic phenotype. Full article

(This article belongs to the Special Issue Staphylococcus spp. in Animals: Resistance to Antimicrobials, Virulence and Genetic Lineages)

13 pages, 6330 KiB

Open AccessArticle

Quantification and Trends of Antimicrobial Use in Commercial Broiler Chicken Production in Pakistan

by Muhammad Umair, Muhammad Farooq Tahir, Riasat Wasee Ullah, Jabir Ali, Naila Siddique, Ayesha Rasheed, Muhammad Akram, Muhammad Usman Zaheer and Mashkoor Mohsin

Antibiotics 2021, 10(5), 598; https://doi.org/10.3390/antibiotics10050598 - 18 May 2021

Cited by 26 | Viewed by 6990

Abstract

Antimicrobial resistance (AMR) is a global health challenge and antimicrobial use (AMU) in the livestock sector has been considered as one of the contributing factors towards the development of AMR in bacteria. This study summarizes the results of a point prevalence survey conducted [...] Read more.

Antimicrobial resistance (AMR) is a global health challenge and antimicrobial use (AMU) in the livestock sector has been considered as one of the contributing factors towards the development of AMR in bacteria. This study summarizes the results of a point prevalence survey conducted to monitor farm-level AMU in commercial broiler chicken farms in Punjab and Khyber Pakhtunkhwa (KPK) provinces of Pakistan. A cross-sectional study was conducted to quantify AMU and to check seasonal variations of AMU in 12 commercial broiler chicken farms (six from each province) during the summer and winter seasons of the year 2020–2021. AMU was recorded using three AMU metrics: kg, mg per population correction unit (mg/PCU), and mg/kg of final flock weight. A total of 22 antimicrobial drugs (348.59 kg) were used for therapeutic or prophylactic purposes in surveyed broiler chicken farms. The total combined AMU for all the broiler chicken farms was 462.57 mg/PCU. The use of most of the antimicrobials increased during winter flocks compared to summer. The top three antimicrobial drugs used during the summer were neomycin (111.39 mg/PCU), doxycycline (91.91 mg/PCU), and tilmicosin (77.22 mg/PCU), whereas doxycycline (196.81 mg/PCU), neomycin (136.74 mg/PCU), and amoxicillin (115.04 mg/PCU) during the winter. Overall, 60% of the antibiotics used in broiler chicken were critically important antimicrobial classes (CIA) for human medicine as characterized by the World Health Organization. Our findings showed high AMU in broiler chicken production and a call for urgent actions to regulate CIA use in food animals in Pakistan. This baseline survey is critical for the design and implementation of a subsequent national level AMU surveys that can include additional farming types, animals’ species, and geographical locations over a longer period of time. Full article

(This article belongs to the Special Issue Usage of Antibiotic in Agriculture and Animal Farming)

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16 pages, 1915 KiB

Open AccessArticle

The Influence of Cellulose-Type Formulants on Anti-Candida Activity of the Tyrocidines

by Yasamin Masoudi, Wilma van Rensburg, Bernice Barnard-Jenkins and Marina Rautenbach

Antibiotics 2021, 10(5), 597; https://doi.org/10.3390/antibiotics10050597 - 18 May 2021

Cited by 3 | Viewed by 2668

Abstract

Candida species are highly adaptable to environmental changes with their phenotypic flexibility allowing for the evasion of most host defence mechanisms. Moreover, increasing resistance of human pathogenic Candida strains has been reported against all four classes of available antifungal drugs, which highlights the [...] Read more.

Candida species are highly adaptable to environmental changes with their phenotypic flexibility allowing for the evasion of most host defence mechanisms. Moreover, increasing resistance of human pathogenic Candida strains has been reported against all four classes of available antifungal drugs, which highlights the need for combinational therapies. Tyrocidines are cyclic antimicrobial peptides that have shown synergistic activity with antifungal drugs such as caspofungin and amphotericin B. However, these cyclodecapeptides have haemolytic activity and cytotoxicity, but they have been used for decades in the clinic for topical applications. The tyrocidines tend to form higher-order structures in aqueous solutions and excessive aggregation can result in variable or diminished activity. Previous studies have shown that the tyrocidines prefer ordered association to celluloses. Therefore, a formulation with soluble cellulose was used to control the oligomer stability and size, thereby increasing the activity against Candida spp. Of the formulants tested, it was found that commercial hydroxy-propyl-methyl cellulose, E10M, yielded the best results with increased stability, increased anti-Candida activity, and improved selectivity. This formulation holds promise in topical applications against Candida spp. infections. Full article

(This article belongs to the Special Issue Therapeutic Use of Antimicrobial Peptides: Joys and Sorrows)

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Examples of cellulose derivatives and Trcs studied with (A) cellulose polymer, (B) core of hydroxyl and methylcellulose derivatives and (C) primary structures of tyrocidine A and C. Full article ">Figure 2
ESMS analysis of the cyclodecapeptides in the Trc mix. The mass spectra were generated with the MaxEnt 3 algorithm in MassLynx 4.01. The top spectrum shows the monomers in the Trc mix and the bottom spectrum shows the homo- and hetero-oligomers that were detected. Full article ">Figure 3
Metabolic inhibition of C. albicans cultures treated by formulated Trc mix at 50 µg/mL (A), 25 µg/mL (B) and 12.5 µg/mL (C). Each data point represents one treated culture. The mean % metabolic inhibition, calculated in terms of growth controls, is shown for each condition with standard deviation (SD). Full article ">Figure 4
Monitoring the influence of the formulation on the oligomerisation concentration of the cyclodecapeptides in the Trc mix. Control (A); Trc mix: E4M (1:4) (B). Fluorescence excitation and emission of the Trc mix (1.25–100 µg/mL) were at 280 and 348 nm, respectively. Three repeats of each preparation are shown via scatter dot plots. Continual readings were taken for the first 60 min with F60/F10 (the reading taken at 60 min/reading taken at 10 min). A sigmoidal curve with a variable slope was fitted to each of the data sets (R2 > 0.99) and the intercept between the slope and the plateau was taken as the critical oligomerisation concentration. Full article ">Figure 5
Representative dose-response curves of the Trc mix alone and the selected fresh 1:4 formulations. The average of four biological repeats (n = 16–30 dose-response curves) shows the activity of the Trc mix and its formulation against C. albicans. Each data point depicts the average and the standard error of the mean (SEM), and the curves were fitted with a sigmoidal curve with a variable slope. Full article ">Figure 6
Comparison of the IC50 µg/mL against C. albicans and C. glabrata of 20-h-matured preparations (1:4 ratio). Data represent the mean of 3 biological repeats and 6–14 technical repeats with SEM. Unpaired Student’s t-test was done on each of the analysed groups, with ** p < 0.01 and *** p < 0.001. Full article ">Figure 7
Comparison of the total fluorescence of the formulated Trc mix at 50 µg/mL and different ratios of cellulose derivatives at 1 h and after 20 h, showing the formulations with 1:1 (m/m) ratio (A), and those with 1:4 (m/m) ratio (C). Each bar represents the mean of 12 preparations and their determinations with SD. The S/N comparison of the respected datasets is shown in (B) and (D). The dotted lines show the comparison with the controls at 1 h and 20 h. Full article ">

19 pages, 5614 KiB

Open AccessArticle

Pan-Resistome Insights into the Multidrug Resistance of Acinetobacter baumannii

by Diego Lucas Neres Rodrigues, Francielly Morais-Rodrigues, Raquel Hurtado, Roselane Gonçalves dos Santos, Daniela Camargos Costa, Debmalya Barh, Preetam Ghosh, Khalid J. Alzahrani, Siomar Castro Soares, Rommel Ramos, Aristóteles Góes-Neto, Vasco Azevedo and Flávia Figueira Aburjaile

Antibiotics 2021, 10(5), 596; https://doi.org/10.3390/antibiotics10050596 - 18 May 2021

Cited by 14 | Viewed by 4799

Abstract

Acinetobacter baumannii is an important Gram-negative opportunistic pathogen that is responsible for many nosocomial infections. This etiologic agent has acquired, over the years, multiple mechanisms of resistance to a wide range of antimicrobials and the ability to survive in different environments. In this [...] Read more.

Acinetobacter baumannii is an important Gram-negative opportunistic pathogen that is responsible for many nosocomial infections. This etiologic agent has acquired, over the years, multiple mechanisms of resistance to a wide range of antimicrobials and the ability to survive in different environments. In this context, our study aims to elucidate the resistome from the A. baumannii strains based on phylogenetic, phylogenomic, and comparative genomics analyses. In silico analysis of the complete genomes of A. baumannii strains was carried out to identify genes involved in the resistance mechanisms and the phylogenetic relationships and grouping of the strains based on the sequence type. The presence of genomic islands containing most of the resistance gene repertoire indicated high genomic plasticity, which probably enabled the acquisition of resistance genes and the formation of a robust resistome. A. baumannii displayed an open pan-genome and revealed a still constant genetic permutation among their strains. Furthermore, the resistance genes suggest a specific profile within the species throughout its evolutionary history. Moreover, the current study performed screening and characterization of the main genes present in the resistome, which can be used in applied research to develop new therapeutic methods to control this important bacterial pathogen. Full article

(This article belongs to the Special Issue Genomic Analysis of Antibiotics Resistance in Pathogens)

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Graphical representation of the global distribution of isolation sites of different strains of Acinetobacter baumannii in a grouped way. The colors represent the sequence types of the strains in this study. The size of the circle indicates the number of isolated strains. Full article ">Figure 2
Phylogenomic tree based on the core genome of 206 Acinetobacter baumannii strains. The colors represent the grouping by sequence type. The method used was maximum likelihood with statistical support of 1000 bootstraps with 1999 genes present in the core genome. Full article ">Figure 3
Representation of the circular genome of the A. baumannii AYE strain as a central genome. The compared strains were grouped and colored. Full article ">Figure 4
Development curve of the pan-genome of Acinetobacter baumannii. Full article ">Figure 5
Graphical representation of the gene distribution by metabolic pathway within each subpartition of the total pan-genome. Only pathways with at least 0.1% of the genes represented in each subpartition of the pan-genome were considered. Full article ">Figure 6
Distribution of the resistance mechanisms related to each antimicrobial found in the database used in the predicted total pan-resistome. Full article ">Figure 7
Distribution of the resistance mechanisms related to each type of action provided by the translated protein. Full article ">

16 pages, 1371 KiB

Open AccessArticle

Colistin Dosing Regimens against Pseudomonas aeruginosa in Critically Ill Patients: An Application of Monte Carlo Simulation

by Van Thi Khanh Nguyen, Preecha Montakantikul, Pramote Tragulpiankit, Jantana Houngsaitong and Mohd Fazli Shuib

Antibiotics 2021, 10(5), 595; https://doi.org/10.3390/antibiotics10050595 - 17 May 2021

Cited by 3 | Viewed by 3640

Abstract

Our aims are to assess various colistin dosing regimens against Pseudomonas aeruginosa (P. aeruginosa) infection in critically ill patients and to propose an appropriate regimen based on microbiological data. A Monte Carlo simulation was performed using the published colistin’s pharmacokinetic parameters [...] Read more.

Our aims are to assess various colistin dosing regimens against Pseudomonas aeruginosa (P. aeruginosa) infection in critically ill patients and to propose an appropriate regimen based on microbiological data. A Monte Carlo simulation was performed using the published colistin’s pharmacokinetic parameters of critically ill patients, the published pharmacodynamic target from a mouse thigh infection model, and the minimum inhibitory concentration (MIC) results from a Vietnamese hospital. The probability of target attainment (PTA) of 80% and cumulative fraction of response (CFR) of 90% were used to evaluate the efficacy of each regimen. Of 121 P. aeruginosa laboratory datasets, the carbapenem-resistant P. aeruginosa (CRPA) and the colistin-resistant P. aeruginosa rates were 29.8% and 0.8%, respectively. MIC_50,90 were both 0.5 mg/L. The simulated results showed that at MIC of 2 mg/L, most regimens could not reach the PTA target, particularly in patients with normal renal function (Creatinine clearance (CrCl) ≥ 80 mL/min). At MIC of 0.5 mg/L and 1 mg/L, current recommendations still worked well. On the basis of these results, aside from lung infection, our study recommends three regimens against P. aeruginosa infection at MIC of 0.5 mg/L, 1 mg/L, and 2 mg/L. In conclusion, higher total daily doses and fractionated colistin dosing regimens could be the strategy for difficult-to-acquire PTA cases, while a less aggressive dose might be appropriate for empirical treatment in settings with low MIC_50/90. Full article

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Minimum inhibitory concentration (MIC) distribution of colistin among P. aeruginosa isolates. Full article ">Figure 2
Probability of target attainment (PTA) achievement in the groups of CrCl 101–130 mL/min (a), 80–100 mL/min (b), 51–19 mL/min (c), 30–50 mL/min (d), 11–29 mL/min (e), 1–10 mL/min (f). The detail of PTA results is found in <a href="#app1-antibiotics-10-00595" class="html-app">Supplementary material Tables S1–S6</a>. LD: loading dose (in mg colistin base activity), MD: maintenance dose (in colistin base activity), US-FDA: Food and Drug Administration of United State, EMA: European Medicines Agency, q12: every 12 h. Full article ">Figure 2 Cont.
Probability of target attainment (PTA) achievement in the groups of CrCl 101–130 mL/min (a), 80–100 mL/min (b), 51–19 mL/min (c), 30–50 mL/min (d), 11–29 mL/min (e), 1–10 mL/min (f). The detail of PTA results is found in <a href="#app1-antibiotics-10-00595" class="html-app">Supplementary material Tables S1–S6</a>. LD: loading dose (in mg colistin base activity), MD: maintenance dose (in colistin base activity), US-FDA: Food and Drug Administration of United State, EMA: European Medicines Agency, q12: every 12 h. Full article ">Figure 2 Cont.
Probability of target attainment (PTA) achievement in the groups of CrCl 101–130 mL/min (a), 80–100 mL/min (b), 51–19 mL/min (c), 30–50 mL/min (d), 11–29 mL/min (e), 1–10 mL/min (f). The detail of PTA results is found in <a href="#app1-antibiotics-10-00595" class="html-app">Supplementary material Tables S1–S6</a>. LD: loading dose (in mg colistin base activity), MD: maintenance dose (in colistin base activity), US-FDA: Food and Drug Administration of United State, EMA: European Medicines Agency, q12: every 12 h. Full article ">

12 pages, 465 KiB

Open AccessArticle

Antimicrobial Activity of Sorghum Phenolic Extract on Bovine Foodborne and Mastitis-Causing Pathogens

by Sydney E. Schnur, Raghavendra G. Amachawadi, Giovanna Baca, Sarah Sexton-Bowser, Davina H. Rhodes, Dmitriy Smolensky, Thomas J. Herald, Ramasamy Perumal, Daniel U. Thomson and Tiruvoor G. Nagaraja

Antibiotics 2021, 10(5), 594; https://doi.org/10.3390/antibiotics10050594 - 17 May 2021

Cited by 11 | Viewed by 3287

Abstract

Antimicrobial resistance in bacterial pathogens associated with bovine mastitis and human foodborne illnesses from contaminated food and water have an impact on animal and human health. Phenolic compounds have antimicrobial properties and some specialty sorghum grains are high in phenolic compounds, and the [...] Read more.

Antimicrobial resistance in bacterial pathogens associated with bovine mastitis and human foodborne illnesses from contaminated food and water have an impact on animal and human health. Phenolic compounds have antimicrobial properties and some specialty sorghum grains are high in phenolic compounds, and the grain extract may have the potential as a natural antimicrobial alternative. The study’s objective was to determine antimicrobial effects of sorghum phenolic extract on bacterial pathogens that cause bovine mastitis and human foodborne illnesses. Bacterial pathogens tested included Escherichia coli, Salmonella Typhimurium, Campylobacter jejuni, Campylobacter coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Klebsiella oxytoca, Staphylococcus aureus, and Enterococcus faecalis. Antibacterial activities of sorghum phenolic extracts were determined by agar-well diffusion assay. Sorghum phenolic extract was added to the wells in concentrations of 0, 100, 200, 500, 1000, or 4000 µg/mL. The control wells did not receive phenolic extract. Plates were incubated for 18–24 h, and the diameter of each zone of inhibition was measured. The results indicated that sorghum phenolic extract had inhibitory effects on Staphylococcus aureus, Enterococcus faecalis, Campylobacter jejuni, and Campylobacter coli. Full article

(This article belongs to the Section Plant-Derived Antibiotics)

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22 pages, 3454 KiB

Open AccessReview

Bacterial Resistance to Antimicrobial Agents

by Manuel F. Varela, Jerusha Stephen, Manjusha Lekshmi, Manisha Ojha, Nicholas Wenzel, Leslie M. Sanford, Alberto J. Hernandez, Ammini Parvathi and Sanath H. Kumar

Antibiotics 2021, 10(5), 593; https://doi.org/10.3390/antibiotics10050593 - 17 May 2021

Cited by 131 | Viewed by 25985

Abstract

Bacterial pathogens as causative agents of infection constitute an alarming concern in the public health sector. In particular, bacteria with resistance to multiple antimicrobial agents can confound chemotherapeutic efficacy towards infectious diseases. Multidrug-resistant bacteria harbor various molecular and cellular mechanisms for antimicrobial resistance. [...] Read more.

Bacterial pathogens as causative agents of infection constitute an alarming concern in the public health sector. In particular, bacteria with resistance to multiple antimicrobial agents can confound chemotherapeutic efficacy towards infectious diseases. Multidrug-resistant bacteria harbor various molecular and cellular mechanisms for antimicrobial resistance. These antimicrobial resistance mechanisms include active antimicrobial efflux, reduced drug entry into cells of pathogens, enzymatic metabolism of antimicrobial agents to inactive products, biofilm formation, altered drug targets, and protection of antimicrobial targets. These microbial systems represent suitable focuses for investigation to establish the means for their circumvention and to reestablish therapeutic effectiveness. This review briefly summarizes the various antimicrobial resistance mechanisms that are harbored within infectious bacteria. Full article

(This article belongs to the Special Issue Antimicrobial Agents Used in Intensive Care Unit)

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16 pages, 2473 KiB

Open AccessArticle

Antifungal and Anti-Inflammatory Potential of Bupleurum rigidum subsp. paniculatum (Brot.) H.Wolff Essential Oil

by Mónica Zuzarte, Pedro M. P. Correia, Jorge M. Alves-Silva, Maria J. Gonçalves, Carlos Cavaleiro, Teresa Cruz and Lígia Salgueiro

Antibiotics 2021, 10(5), 592; https://doi.org/10.3390/antibiotics10050592 - 17 May 2021

Cited by 9 | Viewed by 3321

Abstract

Fungal infections remain a major health concern with aromatic plants and their metabolites standing out as promising antifungal agents. The present study aims to assess, for the first time, the antifungal and anti-inflammatory potential of Bupleurum subsp. paniculatum (Brot.) H.Wolff essential oil from [...] Read more.

Fungal infections remain a major health concern with aromatic plants and their metabolites standing out as promising antifungal agents. The present study aims to assess, for the first time, the antifungal and anti-inflammatory potential of Bupleurum subsp. paniculatum (Brot.) H.Wolff essential oil from Portugal. The oil obtained by hydrodistillation and characterized by GC-MS, showed high amounts of monoterpene hydrocarbons, namely α-pinene (29.0–36.0%), β–pinene (26.1–30.7%) and limonene (10.5–13.5%). The antifungal potential was assessed, according to CLSI guidelines, against several clinical and collection strains. The essential oil showed a broad fungicidal effect being more potent against Cryptococcus neoformans and dermatophytes. Moreover, a significant germ tube inhibition was observed in Candida albicans as well as a disruption of mature biofilms, thus pointing out an effect of the oil against relevant virulent factors. Furthermore, fungal ultrastructural modifications were detected through transmission electron microscopy, highlighting the nefarious effect of the oil. Of relevance, the oil also evidenced anti-inflammatory activity through nitric oxide inhibition in macrophages activated with lipopolysaccharide. In addition, the essential oil’s bioactive concentrations did not present toxicity towards macrophages. Overall, the present study confirmed the bioactive potential of B. rigidum subsp. paniculatum essential oil, thus paving the way for the development of effective drugs presenting concomitantly antifungal and anti-inflammatory properties. Full article

(This article belongs to the Special Issue Antimicrobial Activity of Essential Oils)

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Filamentation percentage of Candida albicans ATCC 10231 treated with different concentrations of Bupleurum rigidum subp. paniculatum essential oil (a) or treated with fluconazole (b). One-way ANOVA followed by Dunnett’s Multiple Comparison Test, ** p < 0.01; *** p < 0.001, compared to control. Full article ">Figure 2
Effect of Bupleurum rigidum subp. paniculatum essential oil (a) and fluoconazole (b) on Candida albicans ATCC 10231 biofilm biomass and viability. One-way ANOVA followed by Dunnett’s multiple comparison test (performed in separate on each feature), * p < 0.05, compared to biofilm mass control; # p < 0.05 and ## p < 0.01, compared to biofilm viability control; arrows indicate MIC value. Full article ">Figure 3
Transmission electron microscopic observations of Candida albicans in the presence of DMSO (a,b) or treated with of 72 µg/mL of Bupleurum rigidum subsp. paniculatum essential oil (c–h). Arrow points to fragments from the cell wall; c—cell wall. Full article ">Figure 4
Transmission electron microscopic observations of Trichophyton rubrum in the presence of DMSO (a,b) or treated with of 576 µg/mL of Bupleurum rigidum subsp. paniculatum essential oil (c–h). Arrow points to fragments from the cell wall; * shows an autophagic structure; c—cell wall, v—vacuoles. Full article ">Figure 5
Effect of Bupleurum rigidum subp. paniculatum essential oil on NO production (a) and cell viability (b). One-way ANOVA followed by Dunnett’s Multiple Comparison Test, ### p < 0.001, compared to control; ** p < 0.01; *** p < 0.001, compared to LPS. EO—essential oil. Full article ">

9 pages, 1283 KiB

Open AccessArticle

Impact of the COVID-19 Pandemic on the Prescribing Patterns of First-Line Antibiotics in English Primary Care: A Longitudinal Analysis of National Prescribing Dataset

by Alisha Zubair Hussain, Vibhu Paudyal and Muhammad Abdul Hadi

Antibiotics 2021, 10(5), 591; https://doi.org/10.3390/antibiotics10050591 - 17 May 2021

Cited by 26 | Viewed by 4661

Abstract

The COVID-19 pandemic has impacted on public access to health services. This study aimed to investigate the impact of COVID-19 pandemic on commonly prescribed first-line antibiotics in English primary care. A secondary analysis of publicly available government data pertaining to primary care prescribing [...] Read more.

The COVID-19 pandemic has impacted on public access to health services. This study aimed to investigate the impact of COVID-19 pandemic on commonly prescribed first-line antibiotics in English primary care. A secondary analysis of publicly available government data pertaining to primary care prescribing was conducted. A list of twenty first-line antibiotics used to treat common infections was developed following the National Institute of Clinical Excellence (NICE) guidelines. All primary care prescription and cost data pertaining to commonly prescribed first-line antibiotics in England between March and September of 2018–2020 were extracted and adjusted for inflation. Analysis suggests prescribing of antibiotics significantly reduced by 15.99% (p = 0.018) and 13.5% (p = 0.002) between March and September 2020 compared with same time period for 2018 and 2019, respectively. The most noticeable decrease in 2020 was noticed for prescribing for meningitis (−62.3%; p = 0.002) followed by respiratory tract infections (−39.13%; p = 0.035), in terms of indications. These results are suggestive of reduced transmission of infections in the community due to national lockdowns, social distancing and hygiene practices. In addition, the impact of reduced face-to-face consultations in general practices needs to be investigated as a potential reason for reduced prescribing. The pandemic also offers an opportunity to rationalize antibiotics use in the community. Full article

(This article belongs to the Special Issue Access, Consumption and Use of Antimicrobials)

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14 pages, 1241 KiB

Open AccessArticle

Coaching Belgian and Dutch Broiler Farmers Aimed at Antimicrobial Stewardship and Disease Prevention

by Nele Caekebeke, Moniek Ringenier, Franca J. Jonquiere, Tijs J. Tobias, Merel Postma, Angelique van den Hoogen, Manon A. M. Houben, Francisca C. Velkers, Nathalie Sleeckx, Arjan Stegeman, Jeroen Dewulf and on behalf of the i-4-1-Health Study Group

Antibiotics 2021, 10(5), 590; https://doi.org/10.3390/antibiotics10050590 - 17 May 2021

Cited by 16 | Viewed by 3489

Abstract

A reduction in antimicrobial use (AMU) is needed to curb the increase in antimicrobial resistance in broiler production. Improvements in biosecurity can contribute to a lower incidence of disease and thereby lower the need for AMU. However, veterinary advice related to AMU reduction [...] Read more.

A reduction in antimicrobial use (AMU) is needed to curb the increase in antimicrobial resistance in broiler production. Improvements in biosecurity can contribute to a lower incidence of disease and thereby lower the need for AMU. However, veterinary advice related to AMU reduction or biosecurity is often not complied with, and this has been linked to the attitudes of farmers. Behavior change promoted by coaching may facilitate uptake and compliance regarding veterinary advice. Thirty broiler farms in Belgium and the Netherlands with high AMU were included in this study for 13 months. For each farmer, the attitude towards AMU reduction was quantified using an adjusted Awareness, Desire, Knowledge, Ability, and Reinforcement (ADKAR^®) change management model, and farm biosecurity was assessed with the Biocheck.UGent^™ tool. Subsequently, farmers were coached to improve disease prevention and antimicrobial stewardship. After the individual coaching of farmers, there was a change in their attitudes regarding AMU, reflected by an increase in ADKAR^® scores. Biosecurity levels improved by around 6% on average, and AMU was reduced by 7% on average without negative effects on performance parameters. Despite these improvements, no significant association could be found between higher ADKAR^® scores and lower AMU. Further investigation into sociological models is needed as a tool to reduce AMU in livestock production. Full article

(This article belongs to the Special Issue Antimicrobial Stewardship in Veterinary Medicine)

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Distribution of (a) external and (b) internal biosecurity levels for participating Belgian (n = 15) and Dutch (n = 13) farms per coaching period. The higher the biosecurity scores are on the y-axis, the better the disease prevention measures in place are. The line within each box represents the median value. The lower and upper boundaries of each box correspond to the 25th and 75th percentiles, respectively. Whiskers below and above each box show the 10th and 90th percentiles. Points below and above the whiskers represent outliers. Full article ">Figure 2
Antimicrobial use per coaching period for the participating broiler farms in Belgium (n = 15) and the Netherlands (n = 13). Antimicrobial use is expressed as the number of days an animal was treated with antimicrobials out of 100 days (treatment incidence (TI) per 100 days). The line within each box represents the median value. The lower and upper boundaries of each box correspond to the 25th and 75th percentiles, respectively. Whiskers below and above each box are the 10th and 90th percentiles. Points below and above the whiskers represent outliers. Full article ">Figure 3
Association pathway according to a linear mixed model. The model included country, coaching period, biosecurity levels, performance parameters (mortality, feed conversion ratio), ADKAR® elements, and antimicrobial use. AMU: antimicrobial use; FCR: feed conversion ratio. Coaching period * country: the interaction between coaching period and country. The model was corrected for farm effects by assigning the latter as a random factor. Estimates were added to the pathway in italics. The non-significant associations with AMU are indicated with a dashed arrow. Full article ">

12 pages, 1452 KiB

Open AccessArticle

Insight on the Structure-to-Activity of Carbosilane Metallodendrimers in the Fight against Staphylococcus aureus Biofilms

by Celia Llamazares, Natalia Sanz del Olmo, Juan Soliveri, F. Javier de la Mata, José Luis Copa-Patiño and Sandra García-Gallego

Antibiotics 2021, 10(5), 589; https://doi.org/10.3390/antibiotics10050589 - 17 May 2021

Cited by 5 | Viewed by 2286

Abstract

Biofilm formation is a critical health concern, involved in most human bacterial infections. Combatting this mechanism, which increases resistance to traditional antibiotics and host immune defences, requires novel therapeutic approaches. The remarkable biocide activity and the monodispersity of carbosilane metallodendrimers make them excellent [...] Read more.

Biofilm formation is a critical health concern, involved in most human bacterial infections. Combatting this mechanism, which increases resistance to traditional antibiotics and host immune defences, requires novel therapeutic approaches. The remarkable biocide activity and the monodispersity of carbosilane metallodendrimers make them excellent platforms to evaluate the impact of different structural parameters on the biological activity. In this work, we explore the influence of iminopyridine ring substituents on the antibacterial activity against planktonic and biofilm Staphylococcus aureus. New families of first-generation Ru(II) and Cu(II) metallodendrimers were synthesised and analysed, in comparison to the non-substituted counterparts. The results showed that the presence of methyl or methoxy groups in meta position to the imine bond decreased the overall positive charge on the metal ion and, subsequently, the activity against planktonic bacteria. However, it seemed a relevant parameter to consider for the prevention of biofilm formation, if they contribute to increasing the overall lipophilicity. An optimum balance of the charge and lipophilicity of the metallodrug, accomplished through structural design, will provide effective biocide agents against bacteria biofilms. Full article

(This article belongs to the Special Issue Antibiotic-Free Antibacterial Strategies Enabled by Nanomaterials)

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14 pages, 1575 KiB

Open AccessArticle

Effects of Antifungal Carriers Based on Chitosan-Coated Iron Oxide Nanoparticles on Microcosm Biofilms

by Anne Caroline Morais Caldeirão, Heitor Ceolin Araujo, Camila Miranda Tomasella, Caio Sampaio, Marcelo José dos Santos Oliveira, Gordon Ramage, Juliano Pelim Pessan and Douglas Roberto Monteiro

Antibiotics 2021, 10(5), 588; https://doi.org/10.3390/antibiotics10050588 - 17 May 2021

Cited by 16 | Viewed by 3698

Abstract

Resistance of Candida species to conventional therapies has motivated the development of antifungal nanocarriers based on iron oxide nanoparticles (IONPs) coated with chitosan (CS). This study evaluates the effects of IONPs-CS as carriers of miconazole (MCZ) or fluconazole (FLZ) on microcosm biofilms. Pooled [...] Read more.

Resistance of Candida species to conventional therapies has motivated the development of antifungal nanocarriers based on iron oxide nanoparticles (IONPs) coated with chitosan (CS). This study evaluates the effects of IONPs-CS as carriers of miconazole (MCZ) or fluconazole (FLZ) on microcosm biofilms. Pooled saliva from two healthy volunteers supplemented with C. albicans and C. glabrata was the inoculum for biofilm formation. Biofilms were formed for 96 h on coverslips using the Amsterdam Active Attachment model, followed by 24 h treatment with nanocarriers containing different concentrations of each antifungal (78 and 156 µg/mL). MCZ or FLZ (156 µg/mL), and untreated biofilms were considered as controls. Anti-biofilm effects were evaluated by enumeration of colony-forming units (CFUs), composition of the extracellular matrix, lactic acid production, and structure and live/dead biofilm cells (confocal laser scanning microscopy-CLSM). Data were analyzed by one-way ANOVA and Fisher LSD’s test (α = 0.05). IONPs-CS carrying MCZ or FLZ were the most effective treatments in reducing CFUs compared to either an antifungal agent alone for C. albicans and MCZ for C. glabrata. Significant reductions in mutans streptococci and Lactobacillus spp. were shown, though mainly for the MCZ nanocarrier. Antifungals and their nanocarriers also showed significantly higher proportions of dead cells compared to untreated biofilm by CLSM (p < 0.001), and promoted significant reductions in lactic acid, while simultaneously showing increases in some components of the extracellular matrix. These findings reinforce the use of nanocarriers as effective alternatives to fight oral fungal infections. Full article

(This article belongs to the Special Issue Synthesis and Biological Activity of Antimicrobial Agents)

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Graphical abstract

Graphical abstract
Full article ">Figure 1
Quantification of colony-forming units (Log10 CFU/mL) of total anaerobes (A), total aerobes (B), mutans streptococci (C), Lactobacillus spp. (D), Candida albicans (E), and Candida glabrata (F) from microcosm biofilms formed for 96 h and treated with different compounds. Biofilms were treated during 24 h with miconazole at 156 µg/mL (MCZ), chitosan (CS)-coated iron oxide nanoparticles (IONPs) carrying MCZ at 78 (IONPs-CS-MCZ78) and 156 µg/mL (IONPs-CS-MCZ156), fluconazole at 156 µg/mL (FLZ) and FLZ-containing nanocarrier at 78 (IONPs-CS-FLZ78) and 156 µg/mL (IONPs-CS-FLZ156). Negative control (NC) represents the biofilm formed for 120 h with pure culture medium. Error bars depict standard deviations of the means. Different lowercase letters represent significant differences among the groups (one-way ANOVA and Fisher LSD’s test; p < 0.05). Comparisons were performed separately for each antifungal, its respective nanocarrier and the NC. Full article ">Figure 2
Mean values (standard deviation) of lactic acid concentration from microcosm biofilms formed for 96 h and treated with different compounds. Biofilms were treated during 24 h with miconazole at 156 µg/mL (MCZ), chitosan (CS)-coated iron oxide nanoparticles (IONPs) carrying MCZ at 78 (IONPs-CS-MCZ78) and 156 µg/mL (IONPs-CS- MCZ156), fluconazole at 156 µg/mL (FLZ), and FLZ-containing nanocarrier at 78 (IONPs-CS-FLZ78) and 156 µg/mL (IONPs-CS-FLZ156). The negative control (NC) represents the biofilm formed for 120 h with pure culture medium. Error bars depict standard deviations of the means. Different lowercase letters represent significant differences among the groups (one-way ANOVA and Fisher LSD’s test; p < 0.05). Comparisons were performed separately for each antifungal, its respective nanocarrier, and the NC. Full article ">Figure 3
Confocal laser scanning microscopy images of 96-h microcosm biofilms treated during 24 h with miconazole (MCZ) at 156 µg/mL (b), chitosan (CS)-coated iron oxide nanoparticles (IONPs) carrying MCZ at 156 µg/mL (c), fluconazole (FLZ) at 156 µg/mL (d), and FLZ-containing nanocarrier at 156 µg/mL (e). The negative control (a) represents the biofilm formed for 120 h with pure culture medium. Red and green fluorescence indicate dead and living cells, respectively. Magnification: 20×. The image (f) represents the percentage of dead cells in relation to the total cells, and different lowercase letters represent significant differences among the groups (one-way ANOVA and Fisher LSD’s test; p < 0.05). Comparisons were performed separately for each antifungal, its respective nanocarrier and negative control. Full article ">

14 pages, 526 KiB

Open AccessArticle

A Pilot Randomised Clinical Trial Comparing a Short-Term Perioperative Prophylaxis Regimen to a Long-Term Standard Protocol in Equine Colic Surgery

by Sabita Diana Stöckle, Dania A. Kannapin, Anne M. L. Kauter, Antina Lübke-Becker, Birgit Walther, Roswitha Merle and Heidrun Gehlen

Antibiotics 2021, 10(5), 587; https://doi.org/10.3390/antibiotics10050587 - 16 May 2021

Cited by 13 | Viewed by 3860

Abstract

Background: For surgical interventions classified as clean or clean-contaminated, including laparotomy, guidelines in human and veterinary medicine recommend a short-term perioperative antibiotic prophylaxis (PAP). In equine colic surgery, however, PAP commonly exceeds 24 h. Objectives: The aim of this study was to compare [...] Read more.

Background: For surgical interventions classified as clean or clean-contaminated, including laparotomy, guidelines in human and veterinary medicine recommend a short-term perioperative antibiotic prophylaxis (PAP). In equine colic surgery, however, PAP commonly exceeds 24 h. Objectives: The aim of this study was to compare a single-shot to a 5-day lasting PAP considering surgical site infections (SSI) and other adverse effects probably associated with the particular antimicrobial regimen. Study design: The study was designed as a randomised non-inferiority pilot study including horses subjected to colic surgery while receiving one of two distinct PAP regimens. Methods: All horses (n = 67) included in the study received the standard physical examination before and after surgery. Colic surgery was performed according to the current standard of the clinic. Horses were randomly assigned to two groups, receiving either the “single-shot” or the “5-day lasting” antibiotic prophylaxis. The “single-shot” group (n = 30) received penicillin and gentamicin only once before and, if needed, during surgery, whereas the “5-day lasting” group (n = 37) received antibiotics for five days. In addition to the standard laboratory examinations, serum amyloid A and fibrinogen were determined preoperatively and during five days after surgery. SSI, postoperative colitis and haemolytic anaemia were classified as postoperative complications potentially related to antibiotic use. Results: The outcome of this preliminary non-inferiority clinical trial showed that the occurrence of postoperative adverse events (i.e., SSI, postoperative colitis and haemolytic anaemia) lacked significant differences between the study groups. Main limitations: The main limitations of this study are the limited group sizes and our inability to blind the study. Conclusions: Single-shot PAP seems to be an alternative approach considering the 5-day lasting protocol commonly used in equine abdominal surgery. However, a proper hygiene management together with a close clinical and laboratory monitoring of the equine patient is indispensable. Full article

(This article belongs to the Special Issue Optimization and Improvement of Veterinary Antimicrobial Treatment to Reduce Antimicrobial Resistance)

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14 pages, 605 KiB

Open AccessEditor’s ChoiceArticle

Impact of an Antimicrobial Stewardship Program on the Incidence of Carbapenem Resistant Gram-Negative Bacilli: An Interrupted Time-Series Analysis

by Teresa López-Viñau, Germán Peñalva, Lucrecia García-Martínez, Juan José Castón, Montserrat Muñoz-Rosa, Ángela Cano, Manuel Recio, José Miguel Cisneros, Elena Pérez-Nadales, José Rumbao Aguirre, Elena García-Martínez, Inmaculada Salcedo, José Ramón del Prado, Carmen de la Fuente, Luis Martínez-Martínez, Irene Gracia-Ahufinger and Julián Torre-Cisneros

Antibiotics 2021, 10(5), 586; https://doi.org/10.3390/antibiotics10050586 - 16 May 2021

Cited by 14 | Viewed by 4605

Abstract

Carbapenem-resistant Gram-negative bacilli (CR-GNB) are a critical public health threat, and carbapenem use contributes to their spread. Antimicrobial stewardship programs (ASPs) have proven successful in reducing antimicrobial use. However, evidence on the impact of carbapenem resistance remains unclear. We evaluated the impact of [...] Read more.

Carbapenem-resistant Gram-negative bacilli (CR-GNB) are a critical public health threat, and carbapenem use contributes to their spread. Antimicrobial stewardship programs (ASPs) have proven successful in reducing antimicrobial use. However, evidence on the impact of carbapenem resistance remains unclear. We evaluated the impact of a multifaceted ASP on carbapenem use and incidence of CR-GNB in a high-endemic hospital. An interrupted time-series analysis was conducted one year before and two years after starting the ASP to assess carbapenem consumption, CR-GNB incidence, death rates of sentinel events, and other variables potentially related to CR-GNB incidence. An intense reduction in carbapenem consumption occurred after starting the intervention and was sustained two years later (relative effect −83.51%; 95% CI −87.23 to −79.79). The incidence density of CR-GNB decreased by −0.915 cases per 1000 occupied bed days (95% CI −1.743 to −0.087). This effect was especially marked in CR-Klebsiella pneumoniae and CR-Escherichia coli, reversing the pre-intervention upward trend and leading to a relative reduction of −91.15% (95% CI −105.53 to −76.76) and −89.93% (95% CI −107.03 to −72.83), respectively, two years after starting the program. Death rates did not change. This ASP contributed to decreasing CR-GNB incidence through a sustained reduction in antibiotic use without increasing mortality rates. Full article

(This article belongs to the Section Antibiotics Use and Antimicrobial Stewardship)

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10 pages, 238 KiB

Open AccessArticle

Clinical and Economic Impact of Community-Onset Urinary Tract Infections Caused by ESBL-Producing Klebsiella pneumoniae Requiring Hospitalization in Spain: An Observational Cohort Study

by Dawid Rozenkiewicz, Erika Esteve-Palau, Mar Arenas-Miras, Santiago Grau, Xavier Duran, Luisa Sorlí, María Milagro Montero and Juan P. Horcajada

Antibiotics 2021, 10(5), 585; https://doi.org/10.3390/antibiotics10050585 - 15 May 2021

Cited by 7 | Viewed by 3085

Abstract

Objective: To analyze the clinical and economic impact of community-onset urinary tract infections (UTIs) caused by extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae requiring hospitalization. Methods: A retrospective cohort study that included all adults with a UTI caused by K. pneumoniae that were [...] Read more.

Objective: To analyze the clinical and economic impact of community-onset urinary tract infections (UTIs) caused by extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae requiring hospitalization. Methods: A retrospective cohort study that included all adults with a UTI caused by K. pneumoniae that were admitted to a tertiary care hospital in Barcelona, Spain, between 2011 and 2015. Demographic, clinical, and economic data were analyzed. Results: One hundred and seventy-three episodes of UTIs caused by K. pneumoniae were studied; 112 were non-ESBL-producing and 61 were ESBL-producing. Multivariate analysis identified ESBL production, acute confusional state associated with UTI, shock, and the time taken to obtain adequate treatment as risk factors for clinical failure during the first seven days. An economic analysis showed differences between ESBL-producing and non-ESBL-producing K. pneumoniae for the total cost of hospitalization per episode (mean EUR 6718 vs EUR 3688, respectively). Multivariate analysis of the higher costs of UTI episodes found statistically significant differences for ESBL production and the time taken to obtain adequate treatment. Conclusion: UTIs caused by ESBL-producing K. pneumoniae requiring hospitalization and the time taken to obtain adequate antimicrobial therapy are associated with worse clinical and economic outcomes. Full article

(This article belongs to the Special Issue Hospital Acquired Infections, Multidrug Resistant (MDR) Bacteria, Alternative Approaches to Antibiotic Therapy)

12 pages, 1221 KiB

Open AccessArticle

Antibacterial Activities of Homemade Matrices Miming Essential Oils Compared to Commercial Ones

by Sofia Oliveira Ribeiro, Véronique Fontaine, Véronique Mathieu, Zhiri Abdesselam, Baudoux Dominique, Stévigny Caroline and Souard Florence

Antibiotics 2021, 10(5), 584; https://doi.org/10.3390/antibiotics10050584 - 14 May 2021

Cited by 5 | Viewed by 2957

Abstract

The increasing bacterial resistance to antibiotics is a worldwide concern. Essential oils are known to possess remarkable antibacterial properties, but their high chemical variability complicates their development into new antibacterial agents. Therefore, the main purpose of this study was to standardize their chemical [...] Read more.

The increasing bacterial resistance to antibiotics is a worldwide concern. Essential oils are known to possess remarkable antibacterial properties, but their high chemical variability complicates their development into new antibacterial agents. Therefore, the main purpose of this study was to standardize their chemical composition. Several commercial essential oils of ajowan (Trachyspermum ammi L.) and thyme (chemotype thymol) (Thymus vulgaris L.) were bought on the market. GC–MS analysis revealed that thyme essential oils have a chemical composition far more consistent than ajowan essential oils. Sometimes thymol was not even the major compound. The most abundant compounds and the homemade mixtures were tested against two Staphylococcus aureus strains. The antibacterial property of β-caryophyllene presented no direct activity against S. aureus LMG 15975, but in association with thymol or carvacrol at equal percentages an MIC of 125 μg/mL was observed. The mixture of those three compounds at equivalent percentages also decreased by 16-fold the MIC of the penicillin V. Against S. aureus LMG 21674, β-caryophyllene presented an MIC of 31.3 μg/mL and decreased by 267-fold the MIC of the penicillin V. These observations led us to question the benefits of using a complex chemical mixture instead of one active compound to fight bacterial resistance. Full article

(This article belongs to the Special Issue Antimicrobial Activity of Essential Oils)

17 pages, 2587 KiB

Open AccessEditor’s ChoiceArticle

Apt (Adenine Phosphoribosyltransferase) Mutation in Laboratory-Selected Vancomycin-Intermediate Staphylococcus aureus

by Reena Lamichhane-Khadka, Santosh Dulal, Jesus A. Cuaron, Richard Pfeltz, Sushim Kumar Gupta, Brian J. Wilkinson and John E. Gustafson

Antibiotics 2021, 10(5), 583; https://doi.org/10.3390/antibiotics10050583 - 14 May 2021

Cited by 3 | Viewed by 3918

Abstract

Comparative genomic sequencing of laboratory-derived vancomycin-intermediate Staphylococcusaureus (VISA) (MM66-3 and MM66-4) revealed unique mutations in both MM66-3 (in apt and ssaA6), and MM66-4 (in apt and walK), compared to hetero-VISA parent strain MM66. Transcriptional profiling revealed that both MM66 VISA [...] Read more.

Comparative genomic sequencing of laboratory-derived vancomycin-intermediate Staphylococcusaureus (VISA) (MM66-3 and MM66-4) revealed unique mutations in both MM66-3 (in apt and ssaA6), and MM66-4 (in apt and walK), compared to hetero-VISA parent strain MM66. Transcriptional profiling revealed that both MM66 VISA shared 79 upregulated genes and eight downregulated genes. Of these, 30.4% of the upregulated genes were associated with the cell envelope, whereas 75% of the downregulated genes were associated with virulence. In concordance with mutations and transcriptome alterations, both VISA strains demonstrated reduced autolysis, reduced growth in the presence of salt and reduced virulence factor activity. In addition to mutations in genes linked to cell wall metabolism (ssaA6 and walK), the same mutation in apt which encodes adenine phosphoribosyltransferase, was confirmed in both MM66 VISA. Apt plays a role in both adenine metabolism and accumulation and both MM66 VISA grew better than MM66 in the presence of adenine or 2-fluoroadenine indicating a reduction in the accumulation of these growth inhibiting compounds in the VISA strains. MM66 apt mutants isolated via 2-fluoroadenine selection also demonstrated reduced susceptibility to the cell wall lytic dye Congo red and vancomycin. Finding that apt mutations contribute to reduced vancomycin susceptibility once again suggests a role for altered purine metabolism in a VISA mechanism. Full article

(This article belongs to the Section Mechanism and Evolution of Antibiotic Resistance)

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Figure 1
Cartoon indicating the location of the Apt A57→V57 mutation in the ß3-domain of Apt expressed by MM66-3 and MM66-4. α-helical regions are depicted by rounded rectangles and β-sheet regions are depicted by arrows (secondary structures predicted by Protscale program <a href="https://web.expasy.org/protscale/" target="_blank">https://web.expasy.org/protscale/</a>, accessed on 25 January 2021). Full article ">Figure 2
Triton X-100 stimulated whole cell autolysis of parent strain MM66, M66-3 and MM66-4. ○, MM66; ☐, MM66-3; Δ, MM66-4. Error bars represent standard deviation (n = 3). Full article ">Figure 3
A. Growth of parent strain MM66 and VISA mutants MM66-3 and MM66-4 in the presence of 5 mM adenine. B. Growth of parent strain MM66 and VISA mutants MM66-3 and MM66-4 in the presence of 5 mM 2-fluoroadenine. Open symbols represent control and closed symbols represent growth with 5 mM adenine (A) or with 5mM 2-fluoroadenine (B). ○, MM66; ☐, MM66-3; and Δ<tt>, </tt>MM66-4. Error bars represent standard deviation (n = 3). Full article ">Figure 4
Growth of parent strain MM66 and FARS mutants MM66-FARS-1 and MM66-FARS-6 in the presence of 5 mM adenine. Open symbols represent control and closed symbols represent growth with 5 mM adenine. ○, MM66; ☐, MM66-FARS-1; and Δ, MM66-FARS-6. Error bars represent standard deviation (n = 3). Full article ">Figure 5
Triton X-100 stimulated whole cell autolysis of parent strain MM66 and FARS mutants. Error bars represent standard deviation (n = 3). Full article ">Figure 6
Growth of diluted cultures of MM66 and MM66 FARS mutants on TSA and TSA + 0.1% Congo red. Full article ">Figure 7
Vancomycin-resistance population analysis of parent strain MM66 and FARS mutants. Full article ">Figure 8
(A). Effects of 2 M NaCl on the growth of parent strain MM66 and VISA strains MM66-3 and MM66-4. (B). Effects of 2 M KCl on the growth of parent strain MM66 and VISA strains MM66-3 and MM66-4. ○, MM66; ☐, MM66-3; and Δ, MM66-4. Error bars represent standard deviation (n = 3). Full article ">

11 pages, 265 KiB

Open AccessFeature PaperArticle

Risk Factors for Amoxicillin-Clavulanate Resistance in Community-Onset Urinary Tract Infections Caused by Escherichia coli or Klebsiella pneumoniae: The Role of Prior Exposure to Fluoroquinolones

by Javier Martínez-Casanova, Silvia Gómez-Zorrilla, Nuria Prim, Agustina Dal Molin, Daniel Echeverría-Esnal, María Pilar Gracia-Arnillas, Elena Sendra, Robert Güerri-Fernández, Xavier Durán-Jordà, Eduardo Padilla, Juan Pablo Horcajada, Santiago Grau and on behalf of the PROA-PSMAR Group

Antibiotics 2021, 10(5), 582; https://doi.org/10.3390/antibiotics10050582 - 14 May 2021

Cited by 8 | Viewed by 3751

Abstract

Background: High rates of amoxicillin-clavulanate (AMC) resistance among Enterobacterales isolated from urinary tract infections (UTIs) were observed in our area. The aim of this study was to identify risk factors associated with AMC resistance in patients with community-onset UTI in emergency departments (EDs). [...] Read more.

Background: High rates of amoxicillin-clavulanate (AMC) resistance among Enterobacterales isolated from urinary tract infections (UTIs) were observed in our area. The aim of this study was to identify risk factors associated with AMC resistance in patients with community-onset UTI in emergency departments (EDs). Methods: A retrospective study was performed of all ED patients with positive urine cultures for Escherichia coli or Klebsiella pneumoniae in a Spanish tertiary-care hospital. Results: 330 urine cultures in all were included: 261 (79.1%) for E. coli and 69 (20.90%) for K. pneumonia. Rates of AMC resistance were 14.94% and 34.78%, respectively. UTI was clinically confirmed in 212 (64.24%) cases. Previous antimicrobial exposure was independently associated with AMC resistance development in E. coli and K. pneumoniae urinary isolates (OR = 2.94, 95% CI = 1.55–5.58). Analyses of infected patients revealed that previous exposure to fluoroquinolones (OR = 3.33, 95% CI = 1.10–10.12, p = 0.034) and to AMC (OR = 5.68, 95% CI = 1.97–16.44, p = 0.001) was significantly associated with isolation of AMC-resistant strains. Conclusions: Prior antibiotic exposure, particularly to AMC or fluoroquinolones, was the only independent risk factor associated with development of AMC resistance in E. coli and K. pneumoniae urinary isolates from patients attending the ED. Full article

15 pages, 2381 KiB

Open AccessArticle

Targeting Internalized Staphylococcus aureus Using Vancomycin-Loaded Nanoparticles to Treat Recurrent Bloodstream Infections

by Danielle Nader, Fajer Yousef, Nicola Kavanagh, Benedict K. Ryan and Steven W. Kerrigan

Antibiotics 2021, 10(5), 581; https://doi.org/10.3390/antibiotics10050581 - 14 May 2021

Cited by 8 | Viewed by 2850

Abstract

The bacterial pathogen Staphylococcus aureus is a leading cause of bloodstream infections, where patients often suffer from relapse despite antibiotic therapy. Traditional anti-staphylococcal drugs display reduced effectivity against internalised bacteria, but nanoparticles conjugated with antibiotics can overcome these challenges. In the present study, [...] Read more.

The bacterial pathogen Staphylococcus aureus is a leading cause of bloodstream infections, where patients often suffer from relapse despite antibiotic therapy. Traditional anti-staphylococcal drugs display reduced effectivity against internalised bacteria, but nanoparticles conjugated with antibiotics can overcome these challenges. In the present study, we aimed to characterise the internalisation and re-emergence of S. aureus from human endothelial cells and construct a new formulation of nanoparticles that target intracellular bacteria. Using an in vitro infection model, we demonstrated that S. aureus invades and persists within endothelial cells, mediated through bacterial extracellular surface adhesion, Fibronectin-binding protein A/B. After internalising, S. aureus localises to vacuoles as determined by transmission electron microscopy. Viable S. aureus emerges from endothelial cells after 48 h, supporting the notion that intracellular persistence contributes to infection relapses during bloodstream infections. Poly lactic-co-glycolic acid nanoparticles were formulated using a water-in-oil double emulsion method, which loaded 10% vancomycin HCl with 82.85% ± 12 encapsulation efficiency. These non-toxic nanoparticles were successfully taken up by cells and demonstrated a biphasic controlled release of 91 ± 4% vancomycin. They significantly reduced S. aureus intracellular growth within infected endothelial cells, which suggests future potential applications for targeting internalised bacteria and reducing mortality associated with bacteraemia. Full article

(This article belongs to the Special Issue Nanoparticles-Based Antimicrobials)

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Figure 1
Staphylococcus aureus internalises into endothelial cells via its surface fibronectin binding proteins. (A) Transmission electron microscopy images of uninfected and S. aureus 8325-4 infected endothelial cells after 48 h were visualised. The side panel demonstrates the internalisation of S. aureus which localises to membrane vacuoles. Scale bars of uninfected and infected at 2 µm. Scale bars of side panel at 500 nm. (B) Dot blot analysis showing FnBP is expressed only in wild-type strain S. aureus 8325-4, and not in FnBP knockout strain S. aureus 8325-4 ΔfnbAB. (C) Endothelial cells were infected with S. aureus 8325-4 over 48 h and intracellular bacteria were quantified as colony forming units, where significant increase in growth was observed (p < 0.01). (D) Primary derived human aortic endothelial cells were infected with the wild-type S. aureus 8325-4 or the mutant S. aureus 8325-4 ΔfnbAB for 1 h. Cells were lysed and total bacteria (external and internal) were quantified using colony forming units. The experiment was repeating using the gentamycin assay method by treating endothelial cells with 100 µg/mL gentamicin HCl for 1 h. Cells were lysed and bacteria were quantified similarly. Wild-type S. aureus significantly internalised (p < 0.0001), yet the strain lacking the fibronectin binding proteins was unable to internalise into the endothelial cells. Results are the mean values ± SEM of three independent experiments using 1-way ANOVA followed by Tukey’s post hoc tests. p < 0.05 indicates statistical significance. ** p < 0.01, **** p< 0.0001. Full article ">Figure 2
Physiochemical characteristics of vancomcyin-loaded PLGA RG 503H nanoparticles. (A) The effect of drug:polymer ratio was evaluated using 10%, 20% and 33% w/w of vancomycin HCl. Particle sizes were significantly different between 10% and 20% vancomycin, and 10% and 33% vancomycin (p < 0.05). The encapsulation efficiency was highest for the 20% formulation; however, considering that small particle sizes are most favourable for nanoparticle use, the 10% vancomycin drug:polymer ratio was chosen for future experiments. (B) The effects of altering the volume of the inner aqueous phase on 10% w/w vancomycin nanoparticles was investigated using 1 mL, 0.5 mL, and 0.3 mL. Reducing the inner aqueous volume did not have a significant difference on average particle size diameter or PDI. However, batches using inner aqueous phase volume of 0.3 mL had the highest encapsulation efficiency of vancomycin and therefore were used for future experiments. (C) The effect of changing the concentration of the surfactant polyvinyl alcohol (PVA) was assessed on 10% w/w vancomycin nanoparticles. Formulations with 5% PVA produced unwanted large particle sizes in contrast to both 1% and 2.5% (p < 0.05). Whilst PVA concentrations had no significant effect on polymer recovery, using 2.5% PVA produced highest encapsulation efficiency and were used for future experiments. Results are the mean values ± SEM of three independent experiments using 1-way ANOVA followed by Tukey’s post hoc tests. p < 0.05 indicates statistical significance. * p < 0.05. Full article ">Figure 2 Cont.
Physiochemical characteristics of vancomcyin-loaded PLGA RG 503H nanoparticles. (A) The effect of drug:polymer ratio was evaluated using 10%, 20% and 33% w/w of vancomycin HCl. Particle sizes were significantly different between 10% and 20% vancomycin, and 10% and 33% vancomycin (p < 0.05). The encapsulation efficiency was highest for the 20% formulation; however, considering that small particle sizes are most favourable for nanoparticle use, the 10% vancomycin drug:polymer ratio was chosen for future experiments. (B) The effects of altering the volume of the inner aqueous phase on 10% w/w vancomycin nanoparticles was investigated using 1 mL, 0.5 mL, and 0.3 mL. Reducing the inner aqueous volume did not have a significant difference on average particle size diameter or PDI. However, batches using inner aqueous phase volume of 0.3 mL had the highest encapsulation efficiency of vancomycin and therefore were used for future experiments. (C) The effect of changing the concentration of the surfactant polyvinyl alcohol (PVA) was assessed on 10% w/w vancomycin nanoparticles. Formulations with 5% PVA produced unwanted large particle sizes in contrast to both 1% and 2.5% (p < 0.05). Whilst PVA concentrations had no significant effect on polymer recovery, using 2.5% PVA produced highest encapsulation efficiency and were used for future experiments. Results are the mean values ± SEM of three independent experiments using 1-way ANOVA followed by Tukey’s post hoc tests. p < 0.05 indicates statistical significance. * p < 0.05. Full article ">Figure 3
Rhodamine B-loaded PLGA RG-503H nanoparticles internalise into human endothelial cells. (A) Human endothelial cells were incubated over 18 hrs (hours) with Rhodamine B-loaded PLGA RG-503H nanoparticles to assess their ability to internalise. Cells were labelled with a GAPDH 14C10 monoclonal antibody for cytoplasm staining, 4′,6-diamidino-2-phenylindole for nucleus staining and Rhodamine B fluorescent dye for nanoparticle staining. Nanoparticles were found to accumulate within the cytoplasm after 18 hrs. Cells were visualised using Zeiss Axio Observer Z1 immunofluorescence microscope. Representative images of each channel alongside a merged image are shown with scale bar = 10 µM. (B) Z-stack images showing localisation of nanoparticles within endothelial cells. Both angles confirm nanoparticles have internalised and do not aggregate or clump on the cell exterior. Full article ">Figure 4
Comparative analyses of formulated nanoparticles. (A) The blank, Rhodamine B-, and 10% vancomycin HCl-loaded nanoparticles were cross-examined to assess the effect of drug encapsulation on particle size, PDI, and zeta potential. There was no significant difference in particle size or PDI between the nanoparticles, and all 3 nanoparticles displayed negative zeta potentials and even size distributions. (B) Transmission electron images of blank (left) and vancomycin HCl- (right) loaded nanoparticles indicate the heterogeneity of each particle size, all displaying smooth spherical shapes. Scale bars at 500 nm. (C) An n vitro drug release profile was calculated using a PUR-A-LYZER mini 6000 dialysis kit. This indicates cumulative percentage increase in vancomycin from nanoparticles over 144 hrs. Results are the mean values ± SEM of three independent experiments using 1-way ANOVA followed by Tukey’s post hoc tests. Full article ">Figure 5
Effectivity of PLGA RG-503H nanoparticles on infected human endothelial cells in vitro. (A) MTT assay was performed to assess toxicity of PLGA RG-503H nanoparticles on human aortic endothelial cells. No significant difference was observed between untreated and treated cells after exposure for 24 h. (B) S. aureus 8325-4-infected endothelial cells were treated with either concentrated vancomycin solution (0.5 mg/mL), or both vancomycin solution and 10% vancomycin-loaded nanoparticles (0.5 µg/mL). Cells were lysed, and the total amount of external and internal S. aureus was quantified then expressed as colony forming units. The control group not exposed to any antibiotics displayed significant bacterial growth. Cells only treated with vancomycin solution were capable of reducing total S. aureus growth (p = 0.0003). Notably, cells treated with both concentrated and encapsulated vancomycin were capable of significantly reducing total S. aureus levels even more so than cells only treated with vancomycin solution (p = 0.045). Results are the mean values ± SEM of three independent experiments using 1-way ANOVA assuming unequal variances followed by Tukey’s post hoc tests. p < 0.05 indicates statistical significance. * p < 0.05, *** p < 0.001. Full article ">Figure 5 Cont.
Effectivity of PLGA RG-503H nanoparticles on infected human endothelial cells in vitro. (A) MTT assay was performed to assess toxicity of PLGA RG-503H nanoparticles on human aortic endothelial cells. No significant difference was observed between untreated and treated cells after exposure for 24 h. (B) S. aureus 8325-4-infected endothelial cells were treated with either concentrated vancomycin solution (0.5 mg/mL), or both vancomycin solution and 10% vancomycin-loaded nanoparticles (0.5 µg/mL). Cells were lysed, and the total amount of external and internal S. aureus was quantified then expressed as colony forming units. The control group not exposed to any antibiotics displayed significant bacterial growth. Cells only treated with vancomycin solution were capable of reducing total S. aureus growth (p = 0.0003). Notably, cells treated with both concentrated and encapsulated vancomycin were capable of significantly reducing total S. aureus levels even more so than cells only treated with vancomycin solution (p = 0.045). Results are the mean values ± SEM of three independent experiments using 1-way ANOVA assuming unequal variances followed by Tukey’s post hoc tests. p < 0.05 indicates statistical significance. * p < 0.05, *** p < 0.001. Full article ">

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