Chemical Characterization and Antimicrobial Properties of the Hydroalcoholic Solution of Echinacea purpurea (L.) Moench. and Propolis from Northern Italy.

In this study, for the first time, the chemical composition of Echinacea purpurea (L.) Moench. and propolis (EAP) hydroalcoholic solution from the Trentino Alto Adige region of northern Italy was investigated by using SPME-GC-MS to describe the volatile content and GC-MS after silylation to detect the non-volatile compounds in the extractable organic matter. The antimicrobial activity of EAP hydroalcoholic solution was evaluated by Minimum Inhibitory Concentration (MIC) determination on 13 type strains, food and clinical isolates. Time Kill Kinetics (TKK) assays and the determination on swimming and swarming motility for 48 h gave more details on the mode of action of EAP solution. The results highlighted the presence of some terpenes and a large number of compounds belonging to different chemical classes. Among these, sugars and organic acids excelled. The EAP hydroalcoholic solution exhibited a strong antimicrobial activity in terms of MIC, with a clear decrease in the cellular load after 48 h. However, the bacterial motility may not be affected by the EAP treatment, displaying a dynamic swarming and swimming motility capacity over time. Given the complexity of chemical profile and the strong antimicrobial effectiveness, the EAP hydroalcoholic solution can be considered a source of bioactive molecules, deserving further investigation for the versatility of application.

Characterization of antibiotic and disinfectant susceptibility in biofilm-forming Acinetobacter baumannii: A focus on environmental isolates.

The clinical role of Acinetobacter baumannii has been highlighted in numerous infectious syndromes with a high mortality rate, due to the high prevalence of multidrug-resistant (MDR) isolates. The treatment and eradication of this pathogen is hindered by biofilm-formation, providing protection from noxious environmental factors and antimicrobials. The aim of this study was to assess the antibiotic susceptibility, antiseptic susceptibility and biofilm-forming capacity using phenotypic methods in environmental A. baumannii isolates. One hundred and fourteen (n = 114) isolates were collected, originating from various environmental sources and geographical regions. Antimicrobial susceptibility testing was carried out using the disk diffusion method, while antiseptic susceptibility was performed using the agar dilution method. Determination of biofilm-forming capacity was carried out using a microtiter-plate based method. Resistance in environmental A. baumannii isolates were highest for ciprofloxacin (64.03%, n = 73), levofloxacin (62.18%, n = 71) and trimethoprim-sulfamethoxazole (61.40%, n = 70), while lowest for colistin (1.75%, n = 2). Efflux pump overexpression was seen in 48.25% of isolates (n = 55), 49.12% (n = 56) were classified as MDR. 6.14% (n = 7), 9.65% (n = 11), 24.65% (n = 28) and 59.65% (n = 68) of isolates were non-biofilm producers, weak, medium, and strong biofilm producers, respectively. No significant differences were observed between non-MDR vs. MDR isolates regarding their distribution of biofilm-producers (P = 0.655). The MIC ranges for the tested antiseptics were as follows: benzalkonium chloride 16-128 µg mL-1, chlorhexidine digluconate 4-128 µg mL-1, formaldehyde 64-256 µg mL-1 and triclosan 2-16 µg mL-1, respectively. The conscientious use of antiseptics, together with periodic surveillance, is essential to curb the spread of these bacteria, and to maintain current infection prevention capabilities.

Multifactorial action of lavender and lavandin oils against filamentous fungi.

AIMS: In this study, five essential oils (EOs) from different species of Lavandula hybrida abrialis, for Lavandula hybrida R.C., Lavandula hybrida 'super A', Lavandula hybrida 'super Z' and Lavandula vera and its hybrids Lavender were evaluated against 26 dust-isolated fungal strains from North Africa. METHODS AND RESULTS: The composition of the different EOs was determined from volume to dry weight. The photochemical analyses were performed via gas chromatography (GC). The cytotoxic effect of five lavender EOs on human epithelial colorectal adenocarcinoma cells (Caco-2) cell line was done. A total of 26 strains of filamentous fungi including Aspergillus spp., Botrytis cinerea, Ceriporia spp., Fusarium spp. and Penicillium glabrum were isolated from sand dust samples via molecular diagnostic tool of PCR. Fungal strains with the lowest minimal lethal concentration (MLC) were Penicillium glabrum, Ceriporia spp. and a strain of Aspergillus spp. CONCLUSIONS: More studies are needed to verify the activity of this EO against more different fungal species, and determine the active ingredients.Significance and impact of study: MIC of the antifungal efficacy relating to EOs was evaluated. The EOs tests showed no cytotoxic effect at very low concentrations, ranging from 0.03% (IC50 0.9132 mg/mL) (L. hybrid Abrialis) to 0.001% (IC50 1.631 mg/mL) (L. hybrid R.C.).

Nanotechnology as a Promising Approach to Combat Multidrug Resistant Bacteria: A Comprehensive Review and Future Perspectives.

The wide spread of antibiotic resistance has been alarming in recent years and poses a serious global hazard to public health as it leads to millions of deaths all over the world. The wide spread of resistance and sharing resistance genes between different types of bacteria led to emergence of multidrug resistant (MDR) microorganisms. This problem is exacerbated when microorganisms create biofilms, which can boost bacterial resistance by up to 1000-fold and increase the emergence of MDR infections. The absence of novel and potent antimicrobial compounds is linked to the rise of multidrug resistance. This has sparked international efforts to develop new and improved antimicrobial agents as well as innovative and efficient techniques for antibiotic administration and targeting. There is an evolution in nanotechnology in recent years in treatment and prevention of the biofilm formation and MDR infection. The development of nanomaterial-based therapeutics, which could overcome current pathways linked to acquired drug resistance, is a hopeful strategy for treating difficult-to-treat bacterial infections. Additionally, nanoparticles' distinct size and physical characteristics enable them to target biofilms and treat resistant pathogens. This review highlights the current advances in nanotechnology to combat MDR and biofilm infection. In addition, it provides insight on development and mechanisms of antibiotic resistance, spread of MDR and XDR infection, and development of nanoparticles and mechanisms of their antibacterial activity. Moreover, this review considers the difference between free antibiotics and nanoantibiotics, and the synergistic effect of nanoantibiotics to combat planktonic bacteria, intracellular bacteria and biofilm. Finally, we will discuss the strength and limitations of the application of nanotechnology against bacterial infection and future perspectives.

Juniperus oxycedrus L. ssp. Essential Oil Microneedles: A Promising Antimicrobial and Wound Healing Activity.

The use of essential oil (EO) in treating infected wounds is still challenging. A lot of effort has been made to make such an application more convenient. Recently, microneedles (MNDs) have been considered as a smart dermal delivery system to overcome the poor absorption and distribution, low bioavailability, and skin penetration of some drugs. The aim of our study is to evaluate the wound healing activity of juniper-EO-loaded MNDs (EO MNDs) against wounds with bacterial and fungal infection. The Polyvinylpyrrolidone (PVP) MNDs were prepared using the gel-filled mold technique and loaded with juniper EO. In vivo models were created and wounds on rats were infected with two clinically isolated bacterial strains Pseudomonas aeruginosa and Staphylococcus aureus. Furthermore, Candida albicans was used to mimic fungal infection and juniper EO MNDs were tested. The obtained results showed an improvement in wound healing which started from the third day after application of the juniper EO MNDs, and at the sixth day post-infection, the treated wounds were significantly smaller than untreated wounds. A complete healing was shown by the 12th day after infection. Furthermore, our cytotoxicity results showed a cytotoxic effect of juniper EO MNDs on epithelial cells, which explained the faster wound healing in rats. Our study showed that juniper EO MNDs represent a novel strategy in EO delivery with minimal invasion. Juniper EO MNDs demonstrated significant antimicrobial activity against both the bacterial strains Pseudomonas aeruginosa and Staphylococcus aureus and against one fungal strain, Candida albicans. Finally, application of juniper EO MNDs exerted promising activity in the treatment and healing of wound infection.

A rare presentation of tubercular osteomyelitis of the foot.

Tuberculosis is a communicable disease that is a major cause of ill health. It is one of the top ten causes of death worldwide and the leading cause of death from a single infectious agent. Its most common clinical presentation is pulmonary involvement. However, approximately 23-30% of tuberculosis patients have extrapulmonary symptoms. A rare (1%) clinical presentation of tuberculosis is foot and ankle infection. This is complicated by the fact that the diagnosis of osteoarticular tuberculosis is difficult. Our case was a 66-year-old multi-pathological pensioner, who, while working in the countryside, had an ankle sprain on the left foot, with a lacerated wound of about 2 cm diameter. The non-endemic area and the negative chest X-ray made the diagnosis extremely complex. However, a multidisciplinary approach with the radiologists and the infectious disease department led to clinical stabilization of the patient. Therefore, awareness and high index of suspicion of the disease is essential and referral to experts should be made if diagnosis is indeterminate despite extensive investigations. The knowledge allows early identification of the disease and prompt therapy in order to avoid long-standing untreated infections which typically cause bone destruction and loss of function. The knowledge is also mandatory for western physicians due to increasing international travel, immigration from less developed countries and increased use of immunosuppressive medications. We believe that this article can bring awareness around osteoarticular tuberculosis and help with improving outcome and eradication of the infection. Level of clinical evidence: 4.

Evaluation of the Antimicrobial and Antivirulent Potential of Essential Oils Isolated from Juniperus oxycedrus L. ssp. macrocarpa Aerial Parts.

As a consequence of the worsening situation with multidrug-resistant (MDR) pathogens and a disparity in the commercialization of novel antimicrobial agents, scientists have been prompted to seek out new compounds with antimicrobial activity from a wide range of sources, including medicinal plants. In the present study, the antibacterial, antifungal, anti-virulence, and resistance-modulating properties of the essential oil from the Sardinian endemic Juniperus oxycedrus L. ssp. macrocarpa aerial parts were evaluated. The GC/MS analysis showed that the main compounds in the oil were α-pinene (56.63 ± 0.24%), limonene (14.66 ± 0.11%), and ß-pinene (13.42 ± 0.09%). The essential oil showed potent antibacterial activity against Gram-positive bacteria (0.25-2 v/v%) and Salmonella spp. (4 v/v%). The strongest fungicidal activity was recorded against Candida auris sessile cells (median FICI was 0.088) but not against C. albicans biofilms (median FICI was 1). The oil showed potent efflux pump inhibitory properties in the case of Staphylococcus aureus and Escherichia coli. The therapeutic potential of Juniperus may be promising for future more extensive research and in vivo tests to develop new drugs against antibiotic and antifungal resistance.

No Correlation between Biofilm-Forming Capacity and Antibiotic Resistance in Environmental Staphylococcus spp.: In Vitro Results.

The production of biofilms is a critical factor in facilitating the survival of Staphylococcus spp. in vivo and in protecting against various environmental noxa. The possible relationship between the antibiotic-resistant phenotype and biofilm-forming capacity has raised considerable interest. The purpose of the study was to assess the interdependence between biofilm-forming capacity and the antibiotic-resistant phenotype in 299 Staphylococcus spp. (S. aureus n = 143, non-aureus staphylococci [NAS] n = 156) of environmental origin. Antimicrobial susceptibility testing and detection of methicillin resistance (MR) was performed. The capacity of isolates to produce biofilms was assessed using Congo red agar (CRA) plates and a crystal violet microtiter-plate-based (CV-MTP) method. MR was identified in 46.9% of S. aureus and 53.8% of NAS isolates (p > 0.05), with resistance to most commonly used drugs being significantly higher in MR isolates compared to methicillin-susceptible isolates. Resistance rates were highest for clindamycin (57.9%), erythromycin (52.2%) and trimethoprim-sulfamethoxazole (51.1%), while susceptibility was retained for most last-resort drugs. Based on the CRA plates, biofilm was produced by 30.8% of S. aureus and 44.9% of NAS (p = 0.014), while based on the CV-MTP method, 51.7% of S. aureus and 62.8% of NAS were identified as strong biofilm producers, respectively (mean OD570 values: S. aureus: 0.779±0.471 vs. NAS: 1.053±0.551; p < 0.001). No significant differences in biofilm formation were observed based on MR (susceptible: 0.824 ± 0.325 vs. resistant: 0.896 ± 0.367; p = 0.101). However, pronounced differences in biofilm formation were identified based on rifampicin susceptibility (S: 0.784 ± 0.281 vs. R: 1.239 ± 0.286; p = 0.011). The mechanistic understanding of the mechanisms Staphylococcus spp. use to withstand harsh environmental and in vivo conditions is crucial to appropriately address the therapy and eradication of these pathogens.

A First Report of Molecular Typing, Virulence Traits, and Phenotypic and Genotypic Resistance Patterns of Newly Emerging XDR and MDR Aeromonas veronii in Mugil seheli.

Aeromonas veronii is associated with substantial economic losses in the fish industry and with food-borne illness in humans. This study aimed to determine the prevalence, antibiogram profiles, sequence analysis, virulence and antimicrobial resistance genes, and pathogenicity of A. veronii recovered from Mugil seheli. A total of 80 fish were randomly gathered from various private farms in Suez Province, Egypt. Subsequently, samples were subjected to clinical, post-mortem, and bacteriological examinations. The retrieved isolates were tested for sequence analysis, antibiogram profile, pathogenicity, and PCR detection of virulence and resistance genes. The prevalence of A. veronii in the examined M. seheli was 22.5 % (18/80). The phylogenetic analyses revealed that the tested A. veronii strains shared high genetic similarity with other A. veronii strains from India, UK, and China. Using PCR it was revealed that the retrieved A. veronii isolates harbored the aerA, alt, ser, ompAII, act, ahp, and nuc virulence genes with prevalence of 100%, 82.9%, 61.7%, 55.3%, 44.7%, 36.17%, and 29.8%, respectively. Our findings revealed that 29.8% (14/47) of the retrieved A. veronii strains were XDR to nine antimicrobial classes and carried blaTEM, blaCTX-M, blaSHV,tetA, aadA1, and sul1 resistance genes. Likewise, 19.1% (9/47) of the obtained A. veronii strains were MDR to eight classes and possessed blaTEM, blaCTX-M, blaSHV,tetA, aadA1, and sul1 genes. The pathogenicity testing indicated that the mortality rates positively correlated with the prevalence of virulence-determinant genes. To our knowledge, this is the first report to reveal the occurrence of XDR and MDR A. veronii in M. seheli, an emergence that represents a risk to public health. Emerging XDR and MDR A. veronii in M. seheli frequently harbored aerA, alt, ser, ompAII, and act virulence genes, and blaTEM, sul1, tetA, blaCTX-M, blaSHV, and aadA1 resistance genes.

Relationship between Biofilm-Formation, Phenotypic Virulence Factors and Antibiotic Resistance in Environmental Pseudomonas aeruginosa

The formation of a protective biofilm by Pseudomonas aeruginosa (PA) is one of the hallmarks of their survival both in vivo and in harsh environmental conditions, thus, biofilm-eradication has relevance from therapeutic perspectives and for infection control. The aim of our study was to investigate the possible relationship between antibiotic resistance, biofilm-forming capacity and virulence factors in n = 166 PA isolates of environmental origin. Antimicrobial susceptibility testing and the phenotypic detection of resistance determinants were carried out using standard protocols. The biofilm-forming capacity of PA was tested using a standardized crystal violet microtiter plate-based method. Motility (swimming, swarming, and twitching) and siderophore production of the isolates were also assessed. Resistance rates were highest for ciprofloxacin (46.98%), levofloxacin (45.18%), ceftazidime (31.92%) and cefepime (30.12%); 19.28% of isolates met the criteria to be classified as multidrug-resistant (MDR). Efflux pump overexpression, AmpC overexpression, and modified Hodge-test positivity were noted in 28.31%, 18.07% and 3.61%, respectively. 22.89% of isolates were weak/non-biofilm producers, while 27.71% and 49.40% were moderate and strong biofilm producers, respectively. Based on MDR status of the isolates, no significant differences in biofilm-production were shown among environmental PA (non-MDR OD570 [mean ± SD]: 0.416 ± 0.167 vs. MDR OD570: 0.399 ± 0.192; p > 0.05). No significant association was observed between either motility types in the context of drug resistance or biofilm-forming capacity (p > 0.05). 83.13% of isolates tested were positive for siderophore production. The importance of PA as a pathogen in chronic and healthcare-associated infections has been described extensively, while there is increasing awareness of PA as an environmental agent in agriculture and aquaculture. Additional studies in this field would be an important undertaking to understand the interrelated nature of biofilm production and antimicrobial resistance, as these insights may become relevant bases for developing novel therapeutics and eradication strategies against PA.

No Correlation between Biofilm Formation, Virulence Factors, and Antibiotic Resistance in Pseudomonas aeruginosa: Results from a Laboratory-Based In Vitro Study.

Pseudomonas aeruginosa (P. aeruginosa) possesses a plethora of virulence determinants, including the production of biofilm, pigments, exotoxins, proteases, flagella, and secretion systems. The aim of our present study was to establish the relationship between biofilm-forming capacity, the expression of some important virulence factors, and the multidrug-resistant (MDR) phenotype in P. aeruginosa. A total of three hundred and two (n = 302) isolates were included in this study. Antimicrobial susceptibility testing and phenotypic detection of resistance determinants were carried out; based on these results, isolates were grouped into distinct resistotypes and multiple antibiotic resistance (MAR) indices were calculated. The capacity of isolates to produce biofilm was assessed using a crystal violet microtiter-plate based method. Motility (swimming, swarming, and twitching) and pigment-production (pyoverdine and pyocyanin) were also measured. Pearson correlation coefficients (r) were calculated to determine for antimicrobial resistance, biofilm-formation, and expression of other virulence factors. Resistance rates were the highest for ceftazidime (56.95%; n = 172), levofloxacin (54.97%; n = 166), and ciprofloxacin (54.64%; n = 159), while lowest for colistin (1.66%; n = 5); 44.04% (n = 133) of isolates were classified as MDR. 19.87% (n = 60), 20.86% (n = 63) and 59.27% (n = 179) were classified as weak, moderate, and strong biofilm producers, respectively. With the exception of pyocyanin production (0.371 ± 0.193 vs. non-MDR: 0.319 ± 0.191; p = 0.018), MDR and non-MDR isolates did not show significant differences in expression of virulence factors. Additionally, no relevant correlations were seen between the rate of biofilm formation, pigment production, or motility. Data on interplay between the presence and mechanisms of drug resistance with those of biofilm formation and virulence is crucial to address chronic bacterial infections and to provide strategies for their management.

Late onset periprosthetic joint infection of the knee caused by Streptococcus anginosus. Case presentation and literature review.

Periprosthetic joint infection (PJI) is one of the most dramatic complications of joint arthroplasty. Although streptococcal bone and joint infections are less common than staphylococcal cases, their role as causative agents of bone and joint remains significant accounting for at least 10% of PJIs. Streptococcus anginosus group (SAG) bacteria are usually found in the normal flora of the urogenital tract, intestinal tract and oropharynx and could cause pyogenic infections to affect brain, lungs and liver. SAG bacteria are uncommonly reported as a cause of osteomyelitis and the involvement of a joint represent a rare event. S. anginosus has been anecdotical related to implant devices infections such as vascular prosthesis or orthopedic implants, however, PJI of the knee has never been fully reported before. We describe the case of a late onset periprosthetic knee infection due to Streptococcus anginosus successfully treated by a two-stage revision arthroplasty and postoperative parenteral Vancomycin, (2 g per day) and Levofloxacin (750 mg per day) for 4 weeks and then oral Levofloxacin for a further 2 weeks.

Relationship between the Biofilm-Forming Capacity and Antimicrobial Resistance in Clinical Acinetobacter baumannii Isolates: Results from a Laboratory-Based In Vitro Study.

The relationship between the multidrug-resistant (MDR) phenotype and biofilm-forming capacity has been a topic of extensive interest among biomedical scientists, as these two factors may have significant influence on the outcomes of infections. The aim of the present study was to establish a possible relationship between biofilm-forming capacity and the antibiotic-resistant phenotype in clinical Acinetobacter baumannii (A. baumannii) isolates. A total of n = 309 isolates were included in this study. Antimicrobial susceptibility testing and the phenotypic detection of resistance determinants were carried out. The capacity of isolates to produce biofilms was assessed using a crystal violet microtiter-plate-based method. Resistance rates were highest for ciprofloxacin (71.19%; n = 220), levofloxacin (n = 68.61%; n = 212), and trimethoprim-sulfamethoxazole (n = 66.02%; n = 209); 42.72% (n = 132) of isolates were classified as MDR; 22.65% (n = 70) of tested isolates were positive in the modified Hodge-test; the overexpression of efflux pumps had significant effects on the susceptibilities of meropenem, gentamicin, and ciprofloxacin in 14.24% (n = 44), 6.05% (n = 19), and 27.51% (n = 85), respectively; 9.39% (n = 29), 12.29% (n = 38), 22.97% (n = 71), and 55.35% (n = 170) of isolates were non-biofilm-producing and weak, moderate, and strong biofilm producers, respectively. A numerical, but statistically not significant, difference was identified between the MDR and non-MDR isolates regarding their biofilm-forming capacity (MDR: 0.495 ± 0.309 vs. non-MDR: 0.545 ± 0.283; p = 0.072), and no association was seen between resistance to individual antibiotics and biofilm formation. Based on numerical trends, MER-resistant isolates were the strongest biofilm producers (p = 0.067). Our study emphasizes the need for additional experiments to assess the role biofilms have in the pathogenesis of A. baumannii infections.

Clinical Outcome and Drug Expenses of Intravitreal Therapy for Diabetic Macular Edema: A Retrospective Study in Sardinia, Italy.

BACKGROUND: Diabetic macular edema (DME) is a leading cause of visual loss in working-age adults. The purpose of this retrospective study was to perform an epidemiological analysis on DME patients treated with intravitreal drugs in a tertiary hospital. The clinical outcome, adverse drug reactions (ADRs), and intravitreal drug expenses were assessed. METHODS: All DME patients treated with Ranibizumab, Aflibercept, Dexamethasone implant, and Fluocinolone Acetonide implant at the Sassari University Hospital, Italy, between January 2017 and June 2020 were included. Central macular thickness (CMT) and best corrected visual acuity (BCVA) were measured. ADRs and drug expenses were analyzed. RESULTS: Two-hundred thirty-one DME patients (mean age: 65 years) received intravitreal agents. Mean CMT and BCVA were 380 µm and 0.5 LogMAR at baseline, 298 µm and 0.44 logMAR after one year (p = 0.04), and 295 µm and 0.4 logMAR at the end of the follow-up period. A total of 1501 intravitreal injections were given; no major ADRs were reported. Treatment cost was €915,000 (€261,429/year). Twenty non-responders to Ranibizumab or Aflibercept were switched to a Dexamethasone implant. In these patients, mean CMT and BCVA were 468 µm and 0.5 LogMar at the time of switching and 362 µm and 0.3 LogMar at the end of the follow-up (p = 0.00014 and p = 0.08, respectively). CONCLUSION: Results confirm that Ranibizumab, Aflibercept, and Dexamethasone implant are effective and safe in DME treatment. A switch to Dexamethasone implant for patients receiving Aflibercept or Ranibizumab with minimal/no clinical benefit should be considered.

Colombian Essential Oil of Ruta graveolens against Nosocomial Antifungal Resistant Candida Strains.

Drug resistance in antifungal therapy, a problem unknown until a few years ago, is increasingly assuming importance especially in immunosuppressed patients and patients receiving chemotherapy and radiotherapy. In the past years, the use of essential oils as an approach to improve the effectiveness of antifungal agents and to reduce antifungal resistance levels has been proposed. Our research aimed to evaluate the antifungal activity of Colombian rue, Ruta graveolens, essential oil (REO) against clinical strains of Candida albicans, Candida parapsilopsis, Candida glabrata, and Candida tropicalis. Data obtained showed that C. tropicalis and C. albicans were the most sensitive strains showing minimum inhibitory concentrations (MIC) of 4.1 and 8.2 µg/mL of REO. Time-kill kinetics assay demonstrated that REO showed a fungicidal effect against C. tropicalis and a fungistatic effect against C. albicans. In addition, an amount of 40% of the biofilm formed by C. albicans was eradicated using 8.2 µg/mL of REO after 1 h of exposure. The synergistic effect of REO together with some antifungal compounds was also investigated. Fractional inhibitory concentration index (FICI) showed synergic effects of REO combined with amphotericin B. REO Lead a disruption in the cellular membrane integrity, consequently resulting in increased intracellular leakage of the macromolecules, thus confirming that the plasma membrane is a target of the mode of action of REO against C. albicans and C. tropicalis.

Antiviral effect of Hornstedtia bella Skornick essential oil from the whole plant against vaccinia virus (VV).

In the prevention of epidemic and pandemic emerging and neglected viral infections, natural products are an important source of lead compounds. Hornstedtia bella Skornickis is a rhizomatous herb growing in the forest of central Vietnam. Hornstedtia bella essential oil (Hb EO) was recently characterised by our group as endowed of antimicrobial activity against Staphylococcus aureus Methicillin-Resistant strains. Here, we describe for the first time the evaluation of Hb EO against a spectrum of viruses responsible for important human diseases. Hb EO resulted active against Vaccinia virus (VV) (EC50 values 80 µg/mL), closely related to variola virus, causative agent of smallpox. Hb EO was able to strongly reduce the viral VV titer in cell-based assay at not cytotoxic concentration and its potential mode of action was characterised by virucidal activity evaluation followed by time-of-addition assay. Furthermore, Hb EO antiviral activity was implemented in a combination study with the mycophenolic acid.

COVID-19, indications for professional football teams and referees training resumption.

These indications were drawn up by the Federal Medical-Scientific Commission (FIGC Commission), supplemented for the necessary time by some experts on the subject; currently they are intended to grant the highest achievable guarantee level to protect the health of players, referees and all professionals involved in case of resumption of collective training (Document dated 18 April 2020). They were designed to minimize the risk of contagion were thus based on the fact that during that phase of SARS-COV-2 virus pandemic (COVID-19) and in the absence of an effective vaccine, the zero-contagion risk did not exist and does not exist to date. Those guidelines have been updated on the basis of ongoing medical-scientific evidence, taking into account the indications given by the Technical-Scientific Committee and the opinion of the Italian Football Federation representatives, during a meeting that took place on May 7 and was transmitted to FIGC on May 11, 2020; these indications are to be considered stringent and binding for the purposes of sport training resumption.

In Vitro Antimicrobial Activity of a New Ophthalmic Solution Containing Hexamidine Diisethionate 0.05% (Keratosept).

PURPOSE: To assess the in vitro antimicrobial activity of a new commercial ophthalmic solution containing hexamidine diisethionate 0.05% (Keratosept). METHODS: Staphylococcus aureus American Type Culture Collection (ATCC 43300), Pseudomonas aeruginosa ATCC 27853, 3 ocular bacterial isolates (1 Staphylococcus epidermidis, 1 S. aureus, and 1 P. aeruginosa), and 5 Candida species were used. The bacterial and fungal isolates were cultured on Columbia blood agar base and Sabouraud-dextrose agar plates, respectively, and incubated overnight at 37°C. Suspensions were prepared in a sterile saline solution with optical density equal to 0.5 McFarland standard (∼10 CFU/mL). Isolate suspensions were made in Keratosept solution to obtain a concentration of 10 CFU/mL. The suspensions were then distributed in conical tubes with a final volume of 1 mL and incubated at 37°C. After 1, 5, 10, 15, 20, 25, 30 minutes, and 24 hours, 10 µL of each suspension was removed, seeded on Columbia blood agar base and Sabouraud-dextrose agar plates and then incubated for 24 hours at 37°C. RESULTS: After 1-minute incubation, there was no growth on the plates seeded with S. aureus ATCC 43300, S. aureus clinical isolate, S. epidermidis clinical isolate, and all 5 Candida species tested. Conversely, Keratosept solution failed to kill the Pseudomonas isolates after 30 minutes exposure and needed 24 hours to eradicate the organisms. CONCLUSIONS: Keratosept ophthalmic solution showed in vitro antimicrobial activity against S. epidermidis, S. aureus, and Candida species. Results suggest that it may be a potential candidate for the treatment of staphylococcal and Candida infections of the ocular surface and have some role in antimicrobial prophylaxis before intravitreal injections.

In vitro antimicrobial activity of a new ophthalmic solution containing povidone-iodine 0.6% (IODIM® ).

PURPOSE: To assess the in vitro antimicrobial activity of a new commercial ophthalmic solution containing povidone-iodine 0.6% (IODIM® ). METHODS: Staphylococcus aureus ATCC 43300, Pseudomonas aeruginosa ATCC 27853, three ocular bacterial isolates (1 S. epidermidis, 1 S. aureus, 1 P. aeruginosa) and five Candida species were used. The bacterial and fungal isolates were cultured on Columbia blood agar base plates and Sabouraud-dextrose agar plates, respectively and incubated overnight at 37°C. Bacterial and fungal suspensions in sterile saline solution were prepared to an optical density equal to 0.5 McFarland standard (approximately 108 CFU/ml). Suspensions of the isolates were made in IODIM® solution to obtain a final concentration of 106 CFU/ml. The suspensions were then distributed in conical tubes in a final volume of 1 ml and incubated at 37°C. At different time-points (1, 5, 10, 15, 20, 25, 30 min and 24 hr), 10 µl of each suspension was removed, seeded on Columbia blood agar base and Sabouraud-dextrose agar plates and then incubated for 24 hr at 37°C. Positive and negative controls were included in all experiments. RESULTS: After 5-min incubation, there was no bacterial growth on any plate. Conversely, IODIM® failed to kill the Candida isolates after 30 min' exposure and needed 24 hr to eradicate the organisms. CONCLUSION: IODIM® ophthalmic solution showed in vitro antimicrobial activity against S. epidermidis, S. aureus, P. aeruginosa and Candida species. Results suggest that it may be a potential candidate for the treatment of ocular surface infections and antimicrobial prophylaxis before intravitreal injections.

Clinical Assessment of New Topical Cream Containing Two Essential Oils Combined with Tretinoin in the Treatment of Acne.

BACKGROUND: Acne is a frequent adolescent disease characterized by inflammatory and non-inflammatory lesions whose topical treatment very often presents adverse phenomena such as irritation or resistance to antibiotics that reduce the patient's compliance. The purpose of this study is to compare a commercial product (Acnatac gel) based on clindamycin-tretinoin (CTG) with a galenic compound containing 2 essential oils (Myrtus communisL. and Origanum vulgare) and tretinoin (MOTC) to evaluate its anti-acne effectiveness and action on the microclimate of the skin. METHODS: Sixty volunteers were randomly divided into an A group using MOTC and a B group, as a positive control, using CTG. The effectiveness was assessed with non-invasive skin analysis (Sebumeter, pH meter, Tewameter and Mexameter) and the counts of the number of lesions, after 15 and 30 days. RESULTS: In both groups, there is a worsening of transepidermal water loss (TEWL) due to tretinoin. MOTC has improved, starting from 15 days of treatment, the papular erythema (p = 0.0329 vs CTG) and has reduced at all times even the rashes of retinoids present in the healthy perilesional skin (p = 0.0329 and p = 0.0017, respectively, at 15 and 30 days). CONCLUSION: MOTC has shown, compared to Acnatac, to have anti-acne efficacy and to possess an anti-inflammatory activity, due to essential oils, able to reduce in vivo erythematous lesions and those induced by retinoids.