Acoustofluidic one-step production of plasmonic Ag nanoparticles for portable paper-based ultrasensitive SERS detection of bactericides.

SERS measurements for monitoring bactericides in dairy products are highly desired for food safety problems. However, the complicated preparation process of SERS substrates greatly impedes the promotion of SERS. Here, we propose acoustofluidic one-step synthesis of Ag nanoparticles on paper substrates for SERS detection. Our method is economical, fast, simple, and eco-friendly. We adopted laser cutting to cut out appropriate paper shapes, and aldehydes were simultaneously produced at the cutting edge in the pyrolysis of cellulose by laser which were leveraged as the reducing reagent. In the synthesis, only 5 µL of Ag precursor was added to complete the reaction, and no reducing agent was used. Our recently developed acoustofluidic device was employed to intensely mix Ag+ ions and aldehydes and spread the reduced Ag nanoparticles over the substrate. The SERS substrate was fabricated in 1 step and 3 min. The standard R6G solution measurement demonstrated the excellent signal and prominent uniformity of the fabricated SERS substrates. SERS detection of the safe concentration of three bactericides, including tetracycline hydrochloride, thiabendazole, and malachite green, from food samples can be achieved using fabricated substrates. We take the least cost, time, reagents, and steps to fabricate the SERS substrate with satisfying performance. Our work has an extraodinary meaning for the green preparation and large-scale application of SERS.

Design, Synthesis, and Target Identification of Novel Phenylalanine Derivatives by Drug Affinity Responsive Target Stability (DARTS) in Xanthomonas oryzae pv Oryzae.

The increasing resistance displayed by plant phytopathogenic bacteria to conventional pesticides has heightened the urgency for the exploration of novel antibacterial agents possessing distinct modes of action (MOAs). In this study, a series of novel phenylalanine derivatives with the unique structure of acylhydrazone dithioether have been designed and synthesized. Bioassay results demonstrated that most target compounds exhibited excellent in vitro antibacterial activity against Xanthomonas oryzae pv oryzae (Xoo) and Xanthomonas axonopodis pv citri (Xac). Among them, the EC50 values of L3, L4, L6, L21, and L22 against Xoo were 7.4, 9.3, 6.7, 8.9, and 5.1 µg/mL, respectively, superior to that of bismerthiazol (BT) and thiodiazole copper (TC) (41.5 and >100 µg/mL); the EC50 values of L3, L4, L5, L6, L7, L8, L20, L21, and L22 against Xac were 5.6, 2.5, 6.2, 4.1, 4.2, 6.4, 6.3, 3.6, and 5.2 µg/mL, respectively, superior to that of BT and TC (43.3 and >100 µg/mL). An unmodified drug affinity responsive target stability (DARTS) technology was used to investigate the antibacterial MOAs of active compound L22, and the 50S ribosomal protein L2 (RL2) as an unprecedented target protein in Xoo cells was first discovered. The target protein RL2 was then expressed and purified. Furthermore, the in vitro interactions by microscale thermophoresis (Kd = 0.050 µM) and fluorescence titration (Ka = 1.4 × 105 M-1) experiments also demonstrated a strong binding force between compound L22 and RL2. Overall, these results not only facilitate the development of novel antibacterial agents but also establish a reliable method for exploring the targets of bactericides.

Virtual screening of natural ligands from five resources to target Ralstonia solanacearum polygalacturonase and endoglucanase.

The present computational study explores novel herbal compounds with potent inhibitory activity against polygalacturonase (PG) and endoglucanase (EG), the extracellular cell wall-degrading enzymes of Ralstonia solanacearum causing crops' bacterial wilt. Phytocompounds of Rosmarinus officinalis L., Coriandrum sativum L., Ocimum basilicum, Cymbopogon citratus, and Thymus vulgaris were first checked to be pharmacokinetically safe and nontoxic. The ligands were then docked to predicted and validated structural models of PG and EG. Molecular dynamic simulations were performed to ensure the dynamic stability of protein-ligand complexes. Carvone and citronellyl acetate were identified to have the best docking energy in binding and inhibiting PG and EG, respectively. In molecular dynamics, root-mean-square deviations of PG-Carvone and EG-Citronellyl acetate complexes indicated the high stability of the ligands in their corresponding cavities. Root-mean-square fluctuations of both proteins indicated unchanged mobility of the binding site residues due to a stable interaction with their ligands. Functional groups on both ligands contributed to the formation of hydrogen bonds with their respective proteins, which were preserved throughout the simulation time. The nonpolar energy component was revealed to significantly contribute to the stability of the docked protein-ligand complexes. Overall, our findings imply the high capability of Carvone and Citronellyl acetate as strong pesticides against the R. solanacearum-caused wilt. This study highlighted the potential of natural ligands in controlling the agricultural bacterial infections, as well as the utility of computational screening techniques in discovering appropriate and potent lead compounds. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03683-z.

Influence of Honey bee Nutritive Jelly Type and Dilution on its Bactericidal Effect on Melissococcus plutonius, the Etiological Agent of European Foulbrood.

To defend themselves against pathogenic microorganisms, honey bees resort to social immunity mechanisms, such as the secretion of antibiotic compounds in the jelly they feed to their larvae. Whereas the bactericidal activity of jelly fed to queen larvae is well studied, little is known about the bioactivity of compositionally different jelly fed to worker larvae. However, the numerous worker larvae are likely to drive the spread of the microorganism and influence its virulence and pathogenesis. Diluted jelly or extracts are mostly used for jelly bioactivity tests, which may bias the evaluation of the pathogen's resistance and virulence. Here, we compared the bactericidal effect of pure and diluted jellies destined for queen and worker larvae on Melissococcus plutonius, the etiological agent of the European foulbrood (EFB) disease of honey bees, and on a secondary invader bacteria, Enterococcus faecalis. We tested three strains of M. plutonius with varying virulence to investigate the association between resistance to antibacterial compounds and virulence. The resistance of the bacteria varied but was not strictly correlated with their virulence and was lower in pure than in diluted jelly. Resistance differed according to whether the jelly was destined for queen or worker larvae, with some strains being more resistant to queen jelly and others to worker jelly. Our results provide a biologically realistic assessment of host defenses via nutritive jelly and contribute to a better understanding of the ecology of M. plutonius and of secondary invaders bacteria in the honey bee colony environment, thus shedding light on the selective forces affecting their virulence and on their role in EFB pathogenesis.

History of Discovery and Environmental Role of Ice Nucleating Bacteria.

The phenomenon of biological ice nucleation that is exhibited by a variety of bacteria is a fascinating phenotype, which has been shown to incite frost damage to frost-sensitive plants and has been proposed to contribute to atmospheric processes that affect the water cycle and earth's radiation balance. This review explores the several possible drivers for the evolutionary origin of the ice nucleation phenotype. These bacteria and the gene required for this phenotype have also been exploited in processes as diverse as reporter gene assays to assess environmentally responsive gene expression in various plant pathogenic and environmental bacteria and in the detection of foodborne human pathogens when coupled with host-specific bacteriophage, whereas ice nucleating bacteria themselves have been exploited in the production of artificial snow for recreation and oil exploration and in the process of freezing of various food products. This review also examines the historical development of our understanding of ice nucleating bacteria, details of the genetic determinants of ice nucleation, and features of the aggregates of membrane-bound ice nucleation protein necessary for catalyzing ice. Lastly, this review also explores the role of these bacteria in limiting the supercooling ability of plants and the strategies and limitations of avoiding plant frost damage by managing these bacterial populations by bactericides, antagonistic bacteria, or cultural control strategies.

Direct Antibiotic Activity of Bacillibactin Broadens the Biocontrol Range of Bacillus amyloliquefaciens MBI600.

Bacillus amyloliquefaciens is considered the most successful biological control agent due to its ability to colonize the plant rhizosphere and phyllosphere where it outgrows plant pathogens by competition, antibiosis, and inducing plant defense. Its antimicrobial function is thought to depend on a diverse spectrum of secondary metabolites, including peptides, cyclic lipopeptides, and polyketides, which have been shown to target mostly fungal pathogens. In this study, we isolated and characterized the catecholate siderophore bacillibactin by B. amyloliquefaciens MBI600 under iron-limiting conditions and we further identified its potential antibiotic activity against plant pathogens. Our data show that bacillibactin production restrained in vitro and in planta growth of the nonsusceptible (to MBI600) pathogen Pseudomonas syringae pv. tomato. Notably, it was also related to increased antifungal activity of MBI600. In addition to bacillibactin biosynthesis, iron starvation led to upregulation of specific genes involved in microbial fitness and competition. IMPORTANCE Siderophores have mostly been studied concerning their contribution to the fitness and virulence of bacterial pathogens. In the present work, we isolated and characterized for the first time the siderophore bacillibactin from a commercial bacterial biocontrol agent. We proved that its presence in the culture broth has significant biocontrol activity against nonsusceptible bacterial and fungal phytopathogens. In addition, we suggest that its activity is due to a new mechanism of action, that of direct antibiosis, rather than by competition through iron scavenging. Furthermore, we showed that bacillibactin biosynthesis is coregulated with the transcription of antimicrobial metabolite synthases and fitness regulatory genes that maximize competition capability. Finally, this work highlights that the efficiency and range of existing bacterial biocontrol agents can be improved and broadened via the rational modification of the growth conditions of biocontrol organisms.

Thermochromic Polyvinyl Alcohol-Iodine Hydrogels with Safe Threshold Temperature for Infectious Wound Healing.

Iodophor (povidone-iodine) has been widely used for antibacterial applications in the clinic. Yet, limited progress in the field of iodine-based bactericides has been achieved since the invention of iodophor. Herein, a blue polyvinyl alcohol-iodine (PAI) complex-based antibacterial hydrogel is explored as a new generation of biocompatible iodine-based bactericides. The obtained PAI hydrogel maintains laser triggered liquefaction, thermochromic, and photothermal features for highly efficient elimination of bacteria. In vitro antibacterial test reveals that the relative bacteria viabilities of Escherichia coli (E.coli) and methicillin-resistant Staphylococcus aureus (MRSA) incubated with PAI hydrogel are only 8% and 3.8%, respectively. Upon single injection of the PAI hydrogel, MRSA-infected open wounds can be efficiently healed in only 5 days, and the healing speed is further accelerated by laser irradiation due to the dynamic interaction between iodine and polyvinyl alcohol, causing up to ∼29% of wound area being closed on day 1. In addition, a safe threshold temperature of skin scald (∼45 °C) emerges for PAI hydrogels because of thermochromic properties, avoiding thermal injuries during irradiation. In addition, no observed toxicity or skin irritation is observed for the PAI hydrogel. This work expands the category of iodine-based bactericides for safe and controllable management of infected wounds.

Octahedral ruthenium and magnesium naringenin 5-alkoxide complexes: NMR analysis of diastereoisomers and in-vivo antibacterial activity against Xylella fastidiosa.

Although many copper-based antimicrobial compounds have been developed to control pathogenic bacteria and fungi in plants and applied for crop protection, there is evidence that several plant pathogens have developed resistance to copper-based antimicrobial compounds, including some Xanthomonas species. Xylella is a bacterial genus belonging to the Xanthomonas family; and X. fastidiosa, which is responsible for citrus variegated chlorosis (CVC) in sweet orange, may develop resistance to one or more copper-based antimicrobials. Because of the time required for the development and approval of new antimicrobials for commercial use, the discovery of novel bactericidal compounds is essential before the development of resistance to the antimicrobials currently in use becomes widespread. Here, we explored the antimicrobial potential of two newly synthesized antimicrobials complexes and one natural compound against X. fastidiosa. Several nuclear magnetic resonance (NMR) assays with high resolution and sensitivity were developed to identify new diastereoisomers in the context of octahedral ruthenium - [Ru(narin)(phen)2]PF6-and magnesium naringenin 5-alkoxide - [Mg(narin)(phen)2]OAc - complexes, obtained in the present work. The NMR assays proved to be powerful tools for the identification of isomers in metal complexes. Moreover, a protocol for the in-vivo determination of the effects of these complexes against X. fastidiosa was developed. The main trunks of X. fastidiosa infected plants were injected with the two complexes as well as with the limonoid azadirachtin using a syringe; the number of bacterial cells in the plants following treatment was estimated via real-time quantitative PCR (qPCR). Importantly, the administration of both complexes and of azadirachtin drastically reduced the number of X. fastidiosa cells in vivo.

Biology and Management of Bacterial Spot of Peppers in Oklahoma.

Bacterial spot, an important disease of bell and chili peppers grown in Oklahoma, is caused by Xanthomonas euvesicatoria. We evaluated isolates from 1995 to 2015 (n = 72) for avirulence alleles and race by assessing hypersensitive responses (HRs) on differentials with resistance genes Bs1, Bs2, Bs3, or Bs4. Most isolates (96%) expressed AvrBs2 (races 1, 3, 7, 8), and only three were virulent on Bs2 (race 6). Chili cultivars, grown locally for capsaicin production, were susceptible to all races. Copper-based spray programs were evaluated on bell pepper hybrids with and without Bs2 resistance from 2008 to 2010 and on bell and jalapeño hybrids with Bs2 or Bs1-2-3 from 2017 to 2018. Bs2 and Bs1-2-3 hybrids generally had lower disease and higher yields (≤21 t/ha) than susceptible entries. Copper reduced disease by 20 to 40% and increased yield by an average of 4 t/ha, but yield responses were not always significant (P = 0.05). In August 2018, disease increased to >50% on Bs2 hybrids but remained low on the Bs1-2-3 hybrid. Despite the breakdown of Bs2 resistance, yields of the Bs2 hybrids were not reduced. Avirulence alleles and race of isolates from susceptible, Bs2, and Bs1-2-3 hybrids at the end of the 2018 trial depended on source plant genetics. AvrBs2 was expressed in 86% of isolates from a susceptible hybrid but not in any isolates from the Bs2 and Bs1-2-3 hybrids. HR resistances effectively protected yield, but their deployment may not be sustainable without cultural practices such as crop rotation that limit pathogen survival and transmission to subsequent crops.

Simultaneous extraction of diverse organic pollutants from environmental water using a magnetic covalent organic framework composite.

Polycyclic aromatic hydrocarbons (PAHs), tetracyclines (TCs), and triphenylmethane dyes (TDs) are common organic pollutants, which may threat the human health or natural microbial communities. In this work, we report a novel multifunctional sorbent based on core-shell magnetic carboxylate-functionalized covalent organic frameworks composites (Fe3O4@COF-COOH) for the simultaneous adsorption of these target analytes via mixed-mode solid phase extraction. The behaviors of the synthetic composite for the adsorption of PAHs, TCs, and TDs were evaluated based on the Freundlich and Langmuir isotherm models. In combination with quantum chemistry calculations, it was found that the multiple interactions including π-π stacking, hydrogen bonding, and electrostatic attractions were existed between COF-COOH and guest molecules. The extraction parameters were optimized, and a novel simultaneous absorption-stepwise desorption (SASD) strategy for the enrichment of PAHs, TCs, and TDs was proposed. By coupling with HPLC-DAD method, the validation results revealed good linearities (R2 ≥ 0.9882) for all analytes. High sensitivity with LODs within the range of 0.003-0.008, 0.02-0.06, and 0.006-0.008 µg L-1 were obtained for PAHs, TCs, and TDs, respectively. High recoveries ranging from 93.6 to 105.8% were obtained with intra-day RSDs of 2.2-6.3% and inter-day RSDs of 3.2-6.5%. The obtained results demonstrated that the proposed SASD strategy using Fe3O4@COF-COOH as sorbents can be extended to other aqueous solutions consisting of trace multi-target analytes.

Mussel-Inspired Fabrication of SERS Swabs for Highly Sensitive and Conformal Rapid Detection of Thiram Bactericides.

As an important sort of dithiocarbamate bactericide, thiram has been widely used for fruits, vegetables and mature crops to control various fungal diseases; however, the thiram residues in the environment pose a serious threat to human health. In this work, silver nanoparticles (AgNPs) were grown in-situ on cotton swab (CS) surfaces, based on the mussel-inspired polydopamine (PDA) molecule and designed as highly sensitive surface-enhanced Raman scattering (SERS) swabs for the conformal rapid detection of bactericide residues. With this strategy, the obtained CS@PDA@AgNPs swabs demonstrated highly sensitive and reproducible Raman signals toward Nile blue A (NBA) probe molecules, and the detection limit was as low as 1.0 × 10-10 M. More critically, these CS@PDA@AgNPs swabs could be served as flexible SERS substrates for the conformal rapid detection of thiram bactericides from various fruit surfaces through a simple swabbing approach. The results showed that the detection limit of thiram residues from pear, grape and peach surfaces was approximately down to the level of 0.12 ng/cm2, 0.24 ng/cm2 and 0.15 ng/cm2 respectively, demonstrating a high sensitivity and excellent reliability toward dithiocarbamate bactericides. Not only could these SERS swabs significantly promote the collection efficiency of thiram residues from irregular shaped matrices, but they could also greatly enhance the analytical sensitivity and reliability, and would have great potential for the on-site detection of residual bactericides in the environment and in bioscience fields.

The in Planta Effective Concentration of Oxytetracycline Against 'Candidatus Liberibacter asiaticus' for Suppression of Citrus Huanglongbing.

Huanglongbing (HLB) or greening currently is the most devastating citrus disease worldwide. The fastidious phloem-colonizing bacterium 'Candidatus Liberibacter asiaticus' (CLas) is the causal agent of citrus HLB in Florida. Bactericides containing the active ingredient oxytetracycline (OTC) have been used in foliar spray to control citrus HLB in Florida since 2016. However, the minimum concentration of OTC required to suppress CLas in planta remains unknown. We developed a new method for evaluating the effects of OTC treatment on CLas titers in infected plants and determined the relationship between OTC residue levels and control levels achieved for CLas using mathematical modeling in greenhouse and field experiments. In both greenhouse and field, OTC spray did not reduce the titers of CLas, and it produced undetectable or mild levels of OTC residue in leaves within 7 days post-application (DPA). In greenhouse, OTC injection at 0.05 g per tree decreased CLas titers to an undetectable level (cycle threshold value ≥ 36.0) from 7 to 30 DPA and produced a residue level of OTC at 0.68 to 0.73 µg/g of fresh tissue over this period. In the field, OTC injection at 0.50 g per tree resulted in the decline of CLas titers by 1.52 log reduction from 14 to 60 DPA, with residue levels of OTC at 0.27 to 0.33 µg/g of fresh tissue. In both trials, a first-order compart model of OTC residue dynamics in leaves of trunk-injected trees was specified for estimating the retention of effective concentrations. Furthermore, nonlinear modeling revealed significant positive correlations between OTC residue levels in leaves and the control levels for CLas achieved. The results suggested that the minimum concentrations of OTC required to suppress CLas populations in planta to below the detection limit are 0.68 and 0.86 µg/g and that the minimum concentrations of OTC required for initial inhibition of CLas growth in planta are ∼0.17 and ∼0.215 µg/g in leaf tissues under greenhouse and field conditions, respectively. This finding highlights that a minimum concentration of OTC should be guaranteed to be delivered to target CLas in infected plants for effective control of citrus HLB.

Magnetic poly(phenylene ethynylene) conjugated microporous polymer microspheres for bactericides enrichment and analysis by ultra-high performance liquid chromatography-tandem mass spectrometry.

The novel multifunction materials for bactericides residue enrichment is attractive. Few materials used in bactericides analysis could simultaneously achieve convenient separation, recycling and trace enrichment. Therefore, a magnetic poly (phenylene ethynylene) conjugated microporous polymer (CMP) microspheres with three-dimensional network structure and covalently built-in Fe3O4 nanoparticles was reported. The structure of dense poly (phenylene ethynylene) frameworks would enhance its conjugation system, which could adsorb bactericides with phenyl structure through π-π stacking. The material could be separated from sample matrix conveniently and recycled due to the super paramagnetism from embedded Fe3O4 nanoparticles. After response surface optimization on extraction and desorption conditions, a method for trace analysis of six bactericides determination was developed by collaborative use of ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The detection limits were in the range of 0.27-3.1 ng / L, and bactericides had been found in vegetables and fruits at concentration among 0.038-0.62 µg/kg. Finally, the method was further applied to analyze the regression equations of bactericide dissipation under natural condition. Effects of different treatment during cleaning process of vegetables and fruits were investigated to seek for the optimal cleaning method. The results revealed that the analytical method based on magnetic CMP which could achieve convenient separation, recycling and trace enrichment, having a great potential for trace analysis of more phenyl pesticides from real samples.

Design, synthesis and biological evaluations of quaternization harman analogues as potential antibacterial agents.

Thirty-three new quaternization harman analogues were synthesized and their antibacterial activity against four Gram-positive and two Gram-negative bacteria were evaluated. The structure-activity relationships were summarized and compounds 4f, 4i, 4l, 4u, 4w, 4x and 5c showed excellent antibacterial activity, low cytotoxicity, good thermal stability and "drug-like" properties. In particular, compound 4x exhibited better bactericidal effect (4-fold superiority against methicillin-resistant Staphylococcus aureus) than standard drugs fosfomycin sodium and ampicillin sodium (minimum inhibitory concentration = 50 nmol/mL). Scanning electron microscopy revealed morphological changes of the bacterial cell surface and the docking evaluation provided a good total score (6.4952) for 4x which is close to the score of ciprofloxacin (6.9723). The results indicated that the quaternization harman analogues might exert their bactericidal effect by damaging bacterial cell membrane and wall, and disrupting the function of type II topoisomerase. In addition, the in vivo antibacterial assay with a protective efficacy of 81.3% further demonstrated the potential of these derivatives as new bactericides and antibiotics.

Fungal Metabolite Antagonists of Plant Pests and Human Pathogens: Structure-Activity Relationship Studies.

Fungi are able to produce many bioactive secondary metabolites that belong to different classes of natural compounds. Some of these compounds have been selected for their antagonism against pests and human pathogens and structure-activity relationship (SAR) studies have been performed to better understand which structural features are essential for the biological activity. In some cases, these studies allowed for the obtaining of hemisynthetic derivatives with increased selectivity and stability in respect to the natural products as well as reduced toxicity in view of their potential practical applications. This review deals with the SAR studies performed on fungal metabolites with potential fungicidal, bactericidal, insecticidal, and herbicidal activities from 1990 to the present (beginning of 2018).

Qualitative Alterations of Bacterial Metabolome after Exposure to Metal Nanoparticles with Bactericidal Properties: A Comprehensive Workflow Based on (1)H NMR, UHPLC-HRMS, and Metabolic Databases.

Metal nanoparticles (NPs) have proven to be more toxic than bulk analogues of the same chemical composition due to their unique physical properties. The NPs, lately, have drawn the attention of researchers because of their antibacterial and biocidal properties. In an effort to shed light on the mechanism through which the bacteria elimination is achieved and the metabolic changes they undergo, an untargeted metabolomic fingerprint study was carried out on Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacteria species. The (1)H NMR spectroscopy, in conjunction with high resolution mass-spectrometry (HRMS) and an unsophisticated data processing workflow were implemented. The combined NMR/HRMS data, supported by an open-access metabolomic database, proved to be efficacious in the process of assigning a putative annotation to a wide range of metabolite signals and is a useful tool to appraise the metabolome alterations, as a consequence of bacterial response to NPs. Interestingly, not all the NPs diminished the intracellular metabolites; bacteria treated with iron NPs produced metabolites not present in the nonexposed bacteria sample, implying the activation of previously inactive metabolic pathways. In contrast, copper and iron-copper NPs reduced the annotated metabolites, alluding to the conclusion that the metabolic pathways (mainly alanine, aspartate, and glutamate metabolism, beta-alanine metabolism, glutathione metabolism, and arginine and proline metabolism) were hindered by the interactions of NPs with the intracellular metabolites.

Antimicrobial cocktails to control bacterial and fungal contamination in Chlamydomonas reinhardtii cultures.

Chlamydomonas reinhardtii is a unicellular green alga widely used for research in photosynthesis, cell cycle regulation, ciliary biogenesis, and other physiological processes. Sterile cultures are needed for these studies, but contamination from bacteria and fungi occurs frequently. Although the One-shot Solution cocktail consisting of carbendazim, ampicillin, and cefotaxime has been developed for removing these contaminants from algal cultures, it is not always effective. Here we report two new antimicrobial cocktails for treating mixed bacterial and fungal contamination of Chlamydomonas cultures. A combination of the bactericide nalidixic acid with one of two fungicides, azoxystrobin or tebuconazole, was more effective than the One-shot Solution cocktail. In some of our tests, we find that alternating use of our new cocktails with One-shot Solution is needed to remove obstinate contaminants.

Antibacterial effect of water-soluble chitosan on representative dental pathogens Streptococcus mutans and Lactobacilli brevis

Dental caries is still a major oral health problem in most industrialized countries. The development of dental caries primarily involves Lactobacilli spp. and Streptococcus mutans. Although antibacterial ingredients are used against oral bacteria to reduce dental caries, some reports that show partial antibacterial ingredients could result in side effects. OBJECTIVES: The main objective is to test the antibacterial effect of water-soluble chitosan while the evaluation of the mouthwash appears as a secondary aim. MATERIAL AND METHODS: The chitosan was obtained from the Application Chemistry Company (Taiwan). The authors investigated the antibacterial effects of water-soluble chitosan against oral bacteria at different temperatures (25-37ºC) and pH values (pH 5-8), and evaluated the antibacterial activities of a self-made water-soluble chitosan-containing mouthwash by in vitro and in vivo experiments, and analyzed the acute toxicity of the mouthwashes. The acute toxicity was analyzed with the pollen tube growth (PTG) test. The growth inhibition values against the logarithmic scale of the test concentrations produced a concentrationresponse curve. The IC50 value was calculated by interpolation from the data. RESULTS: The effect of the pH variation (5-8) on the antibacterial activity of water-soluble chitosan against tested oral bacteria was not significant. The maximal antibacterial activity of water-soluble chitosan occurred at 37ºC. The minimum bactericidal concentration (MBC) of water-soluble chitosan on Streptococcus mutans and Lactobacilli brevis were 400 µg/mL and 500 µg/mL, respectively. Only 5 s of contact between water-soluble chitosan and oral bacteria attained at least 99.60% antibacterial activity at a concentration of 500 µg/mL. The water-soluble chitosan-containing mouthwash significantly demonstrated antibacterial activity that was similar to that of commercial mouthwashes (>99.91%) in both in vitro and in vivo experiments. In addition, the alcohol-free mouthwash exhibited no cytotoxicity and no oral stinging. To the best of our knowledge, this was the first study to combine in vitro and in vivo investigations to analyze the antibacterial properties of water-soluble chitosan-containing mouthwash. CONCLUSIONS: This study illustrated that water-soluble chitosan may be a viable alternative to commercial mouthwashes in the future.

Evaluation of silicon oil on bacterial growth / Avaliação dos efeitos do óleo de silicone no crescimento bacteriano

PURPOSE: To analyze the antimicrobial properties of silicon oil (Óleo de Silicone®, Ophthalmos, Brazil) on in vitro bacterial growth of different microorganisms related to endophthalmitis. METHODS: The following microorganisms were analyzed: (1) Pseudomonas aeruginosa (ATCC 27583); (2) Escherichia coli (ATCC 25922); (3) Staphylococcus aureus (ATCC 25923); (4) Staphylococcus epidermidis (ATCC 12228); (5) Candida albicans (ATCC 10231); (6) Klebsiella pneumoniae (ATCC 13883); and (7) Streptococcus pneumoniae (ATCC 49619). The plates were incubated at 35 ± 2ºC and its growth examined after 24 hours. An empty disk was placed in the center of each plate as a control. RESULTS: No inhibition halos were verified in any of the plates containing the four different concentrations of the bacterial inocula. CONCLUSIONS: The silicon oil 1000 cps does not have any effect on bacterial growth of any of the studied microrganisms.

OBJETIVO: Analisar as propriedades antimicrobianas do óleo de silicone (Óleo de Silicone®, Ophthalmos, Brazil) no crescimento in vitro de diferentes microrganismos relacionados à endoftalmite. MÉTODOS: Os seguintes microrganismos foram analisados: (1) Pseudomonas aeruginosa (ATCC 27583); (2) Escherichia coli (ATCC 25922); (3) Staphylococcus aureus (ATCC 25923); (4) Staphylococcus epidermidis (ATCC 12228); (5) Candida albicans (ATCC 10231); (6) Klebsiella pneumoniae (ATCC 13883); and (7) Streptococcus pneumoniae (ATCC 49619). As placas foram incubadas à temperatura de 35 ± 2ºC e o seu crescimento examinado após 24 horas. Um disco de papel filtro neutro, sem óleo de silicone, foi posicionado no centro de cada placa como controle. RESULTADOS: Não foram encontrados halos de inibição em nenhuma das placas contendo as diferentes concentrações de inóculo bacteriano estudadas. CONCLUSÕES: O Óleo de Silicone® 1000 cps não apresenta efeito no crescimento bacteriano de nenhum dos microrganismos estudados.

Estudo in vitro da ação bactericida e/ou bacteriostática da papaína / Study in vitro of bactericidal and/or bacteriostatic action of the papaíne

OBJETIVO: avaliar a atividade bactericida e/ou bacteriostática de soluções de papaína, em diferentes volumes, à 2%, 4%, 6%, 8%, 10%, 16% e 20% em culturas de S. aureus, P. aeruginosa, E. faecalis, K. pneumoniae e S. typhi, obtidas e identificadas de materiais biológicos provenientes de pacientes atendidos em um laboratório particular de Belém - Pará. MÉTODO: colhidas amostras de cada uma das colônias de bactérias e realizado o repique das mesmas para tubos de ensaio contendo caldo Casoy, até obter-se uma suspensão com turbidez correspondente a 0,5% na escala de Mac Farland, denominada suspensão-mãe. Foram, então, preparados 08 tubos contendo 1,8ml de caldo Casoy, um grupo para cada espécie de bactéria e, em seguida, feito o repique de 200?l da suspensão-mãe em cada tubo. Posteriormente, adicionou-se 200?l, 400?l, 500?l, 1ml ou 2ml de solução de papaína nas concentrações a serem analisadas. Os controles foram realizados somente com as suspensões bacterianas, sem papaína, incubados a 37ºC por 24 horas. Após esta incubação foi feito o repique de cada tubo para placas de Petri contendo Ágar Cled e incubação a 37ºC por 24 horas. RESULTADOS: evidenciou-se crescimento bacteriano em todas as placas semeadas. CONCLUSÃO: não foi observado efeito bactericida ou bacteriostático, in vitro, associado à papaina.

OBJECTIVES: evaluate the bactericidal and/or bacteriostatic activities of 2, 4, 6, 8, 10, 16 and 20% solutions of papaine on cultures of S. aureus, P. aeruginosa, E. faecalis, K. pneumoniae and S. typhi, obtained from biological samples collected from patients attended at the laboratory in Belém, Pará. METHODS: the previously-isolated bacteria were grown in test tubes containing Casoy medium broth, till the broth attained a turbidity of 0.5% on the Mac Farland scale, to form the stock suspensions. Then, 200?l of the stock suspension of each bacteria was inoculated into each of eight test tubes, each containing 1.8 ml of Casoy broth. Various volumes (200?l, 400?l, 500?l, 1ml or 2ml) of each of the papaine solutions were added to the inoculated broth in the tubes. Controls consisted of bacteria-inoculated broth, without papaine. All the tubes were incubated at 37ºC for 24 hours. After incubation, the bacteria from the tubes were inoculated onto Petri dishes containing Cled agar and incubated at 37ºC for 24 hours. RESULTS: was evidenced bacteria grew in all of the inoculated plates. CONCLUSIONS: no bactericidal or bacteriostatic effects, in vitro, associated papaine.