Emanuela Di Campli - Academia.edu (original) (raw)

Papers by Emanuela Di Campli

Research paper thumbnail of Pistacia vera L. oleoresin and levofloxacin is a synergistic combination against resistant Helicobacter pylori strains

Scientific Reports, Mar 15, 2019

The increasing multidrug resistance in Helicobacter pylori, also correlated to its biofilm-formin... more The increasing multidrug resistance in Helicobacter pylori, also correlated to its biofilm-forming ability, underlines the need to search novel strategies to improve the eradication rate. Natural compounds are proposed as antibiotic-resistant-breakers capable to restore the efficacy of conventional drugs. Aim of this work was to evaluate the capability of Pistacia vera L. oleoresin (ORS) to synergize with levofloxacin (LVX) against resistant H. pylori strains. The antimicrobial activity of P. vera L. ORS and LVX and their combinations was determined by MIC/MBC (in neutral and acidic environments) and checkerboard tests. The anti-biofilm effect was determined by biomass quantification. In vivo Galleria mellonella model was used to confirm in vitro data. Pistacia vera L. ORS and LVX MICs ranged respectively from 780 to 3120 mg/l and from 0.12 to 2.00 mg/l, at pH 7.0 and 5.5. MBCs were similar to MICs. Pistacia vera L. ORS was able to synergize with LVX, restoring its effectiveness in LVX resistant strains. Pistacia vera L. ORS, LVX and their synergistic combinations displayed significant biofilm reduction. Pistacia vera L. ORS and LVX, showed protective effect against H. pylori infection on G. mellonella (62% and 63% of survival, respectively). Pistacia vera L. ORS can be considered a promising potentiator to restore the effectiveness of LVX tackling the H. pylori antibiotic resistance phenomenon. Helicobacter pylori is a gastroduodenal pathogen that plays an important role in the pathogenesis of chronic gastritis, peptic ulcer, gastric adenocarcinoma and MALT (mucosa-associated lymphoid tissue) lymphoma. H. pylori infection is difficult to eradicate and requires the combination of different drugs such as clarithromycin, levofloxacin (LVX), amoxicillin, metronidazole, tetracycline and proton pump inhibitor 1-3. The increase of antimicrobial resistance and the failure of therapeutic regimens, strongly underline the need to find novel strategies to enhance the eradication rate, also considering the capability of H. pylori to grow in biofilm mode 4-6. Moreover, the H. pylori antimicrobial resistance profiles vary in different geographic areas, therefore the selection of therapeutic regimens needs to be adjusted according to local resistance patterns, if available 7-10. For all these reasons, the search for alternative and effective new therapeutic schemes is important and urgent 11-14. New approaches to tackle the infections related to multidrug resistant (MDR) bacteria are recently proposed in the search for antibiotic-resistance-breakers capable to synergize with conventional drugs restoring their effectiveness 15-17. To this regards, much interest has been revived in the study of antimicrobial/antivirulence effects of formulations based on medicinal plants 15,18,19. Several plants produce a variety of secondary metabolites such as phenolics, terpenoids and alkaloids, which possess a wide spectrum of biological activities, including the antibacterial ones 14,19-21. They interact with the lipidic bilayer of the cytoplasmic membrane, membrane proteins and enzymes involved in the synthesis of macromolecules, causing increased permeability, loss of proton-motive force and cellular material 14 .

Research paper thumbnail of Microbial Contamination of Smartphone Touchscreens of Italian University Students

Current Microbiology, Dec 15, 2017

In this study, the microbial contamination of smartphones from Italian University students was an... more In this study, the microbial contamination of smartphones from Italian University students was analyzed. A total of 100 smartphones classified as low, medium, and high emission were examined. Bacteria were isolated on elective and selective media and identified by biochemical tests. The mean values of cfu/cm 2 were 0.79 ± 0.01; in particular, a mean of 1.21 ± 0.12, 0.77 ± 0.1 and 0.40 ± 0.10 cfu/cm 2 was present on smartphones at low, medium, and high emission, respectively. The vast majority of identified microorganisms came from human skin, mainly Staphylococci, together with Gram-negative and positive bacilli and yeasts. Moreover, the main isolated species and their mixture were exposed for 3 h to turned on and off smartphones to evaluate the effect of the electromagnetic wave emission on the bacterial cultivability, viability, morphology, and genotypic profile in respect to the unexposed broth cultures. A reduction rate of bacterial growth of 79 and 46% was observed in Staphylococcus aureus and Staphylococcus epidermidis broth cultures, respectively, in the presence of turned on smartphone. No differences in viability were observed in all detected conditions. Small colony variants and some differences in DNA fingerprinting were detected on bacteria when the smartphones were turned on in respect to the other conditions. The colonization of smartphones was limited to human skin microorganisms that can acquire phenotype and genotypic modifications when exposed to microwave emissions.

Research paper thumbnail of Helicobacter pylori Dormant States Are Affected by Vitamin C

International Journal of Molecular Sciences

Helicobacter pylori colonizes human gastric mucosa, overcoming stressful conditions and entering ... more Helicobacter pylori colonizes human gastric mucosa, overcoming stressful conditions and entering in a dormant state. This study evaluated: (i) H. pylori’s physiological changes from active to viable-but-non-culturable (VBNC) and persister (AP) states, establishing times/conditions; (ii) the ability of vitamin C to interfere with dormancy generation/resuscitation. A dormant state was induced in clinical MDR H. pylori 10A/13 by: nutrient starvation (for VBNC generation), incubating in an unenriched medium (Brucella broth) or saline solution (SS), and (for AP generation) treatment with 10xMIC amoxicillin (AMX). The samples were monitored after 24, 48, and 72 h, 8–14 days by OD600, CFUs/mL, Live/Dead staining, and an MTT viability test. Afterwards, vitamin C was added to the H. pylori suspension before/after the generation of dormant states, and monitoring took place at 24, 48, and 72 h. The VBNC state was generated after 8 days in SS, and the AP state in AMX for 48 h. Vitamin C reduced...

Research paper thumbnail of Prebiotic Combinations Effects on the Colonization of Staphylococcal Skin Strains

Microorganisms

Background: An unbalanced skin microbiota due to an increase in pathogenic vs. commensal bacteria... more Background: An unbalanced skin microbiota due to an increase in pathogenic vs. commensal bacteria can be efficiently tackled by using prebiotics. The aim of this work was to identify novel prebiotic combinations by exerting species-specific action between S. aureus and S. epidermidis strains. Methods: First, the antimicrobial/antibiofilm effect of Xylitol-XYL and Galacto-OligoSaccharides–GOS combined with each other at different concentrations (1, 2.5, 5%) against S. aureus and S. epidermidis clinical strains was evaluated in time. Second, the most species-specific concentration was used to combine XYL with Fructo-OligoSaccharides–FOS, IsoMalto-Oligosaccharides–IMO, ArabinoGaLactan–LAG, inulin, dextran. Experiments were performed by OD600 detection, biomass quantification and LIVE/DEAD staining. Results: 1% XYL + 1% GOS showed the best species-specific action with an immediate antibacterial/antibiofilm action against S. aureus strains (up to 34.54% ± 5.35/64.68% ± 4.77) without a re...

Research paper thumbnail of Near-infrared NIR irradiation and sodium hypochlorite: An efficacious association to counteract the Enterococcus faecalis biofilm in endodontic infections

Journal of Photochemistry and Photobiology B: Biology, 2020

, et al., Near-infrared NIR irradiation and sodium hypochlorite: An efficacious association to co... more , et al., Near-infrared NIR irradiation and sodium hypochlorite: An efficacious association to counteract the Enterococcus faecalis biofilm in endodontic infections,

Research paper thumbnail of Graphene Oxide affects Staphylococcus aureus and Pseudomonas aeruginosa dual species biofilm in Lubbock Chronic Wound Biofilm model

Scientific Reports, 2020

Chronic wound management becomes a complex procedure because of the persistence of forming biofil... more Chronic wound management becomes a complex procedure because of the persistence of forming biofilm pathogens that do not respond to antimicrobial treatment. The aim of this paper is to detect the Graphene Oxide-GO effect on Staphylococcus aureus and Pseudomonas aeruginosa dual species wound biofilm in Lubbock Chronic Wound Biofilm-LCWB model. LCWB is a recognized model that mimics the spatial microbial colonization into chronic wounds and reproduces the wound and its clot. Staphylococcus aureus PECHA 10 and P. aeruginosa PECHA 4, are the pathogens used in the study. The GO effect on both in forming and mature biofilms, is detected by the evaluation of the CFU/mg reduction, the cell viability and ultrastructural analysis of the treated LCWBs. Graphene Oxide, at 50 mg/l, shows a significant antibiofilm effect in forming and mature LCWBs. In particular, during the biofilm formation, GO reduces the S. aureus and P. aeruginosa growth of 55.05% ± 4.73 and 44.18% ± 3.91 compared to the con...

Research paper thumbnail of Searching for New Tools to Counteract the Helicobacter pylori Resistance: The Positive Action of Resveratrol Derivatives

Antibiotics, 2020

The drug-resistance phenomenon in Helicobacter pylori underlines the need of novel strategies to ... more The drug-resistance phenomenon in Helicobacter pylori underlines the need of novel strategies to improve the eradication rate including alternative treatments combining antibiotic and non-antibiotic compounds with synergistic action. In this study, the antibacterial (MIC/MBC) and anti-virulence effects (biofilm reduction and swarming motility inhibition) of resveratrol-RSV and new synthetized RSV-phenol derivatives, with a higher bioavailability, alone and combined with levofloxacin-LVX were evaluated against resistant H. pylori clinical strains. The experiments were confirmed in vivo using the Galleria mellonella model. Among the studied RSV derivatives, RSV-3 and RSV-4 possessed higher antibacterial activity with respect to RSV (MICs from 6.25 to 200 µg/mL and from 3.12 to 200 µg/mL, respectively). RSV, RSV-3, and RSV-4 were able to synergize with LVX restoring its effect in two out of seven clinical resistant strains tested for the study. RSV, RSV-3, and RSV-4, alone and with LVX...

Research paper thumbnail of Antimicrobial and Antibiofilm Efficacy of Graphene Oxide against Chronic Wound Microorganisms

Antimicrobial Agents and Chemotherapy, 2018

Chronic wounds represent an increasing problem worldwide. Graphene oxide (GO) has been reported t... more Chronic wounds represent an increasing problem worldwide. Graphene oxide (GO) has been reported to exhibit strong antibacterial activity toward both Gram-positive and Gram-negative bacteria. The aim of this work was to investigate the in vitro antimicrobial and antibiofilm efficacy of GO against wound pathogens. Staphylococcus aureus PECHA 10, Pseudomonas aeruginosa PECHA 4, and Candida albicans X3 clinical isolates were incubated with 50 mg/liter of GO for 2 and 24 h to evaluate the antimicrobial effect. Optical and atomic force microscopy images were performed to visualize the effect of GO on microbial cells. Moreover, the antibiofilm effect of GO was tested on biofilms, both in formation and mature. Compared to the respective time controls, GO significantly reduced the S. aureus growth both at 2 and 24 h in a time-dependent way, and it displayed a bacteriostatic effect in respect to the GO t = 0; an immediate (after 2 h) slowdown of bacterial growth was detected for P. aeruginosa...

Research paper thumbnail of Enterococcus hirae biofilm formation on hospital material surfaces and effect of new biocides

Environmental Health and Preventive Medicine, 2017

Background: Nowadays, the bacterial contamination in the hospital environment is of particular co... more Background: Nowadays, the bacterial contamination in the hospital environment is of particular concern because the hospital-acquired infections (HAIs), also known as nosocomial infections, are responsible for significant morbidity and mortality. This work evaluated the capability of Enterococcus hirae to form biofilm on different surfaces and the action of two biocides on the produced biofilms. Methods: The biofilm formation of E. hirae ATCC 10541 was studied on polystyrene and stainless steel surfaces through the biomass quantification and the cell viability at 20 and 37°C. The effect of LH IDROXI FAST and LH ENZYCLEAN SPRAY biocides on biomasses was expressed as percentage of biofilm reduction. E. hirae at 20 and 37°C produced more biofilm on the stainless steel in respect to the polystyrene surface. The amount of viable cells was greater at 20°C than with 37°C on the two analyzed surfaces. Biocides revealed a good anti-biofilm activity with the most effect for LH ENZYCLEAN SPRAY on polystyrene and stainless steel at 37°C with a maximum biofilm reduction of 85.72 and 86.37%, respectively. Results: E. hirae is a moderate biofilm producer depending on surface material and temperature, and the analyzed biocides express a remarkable antibiofilm action. Conclusion: The capability of E. hirae to form biofilm can be associated with its increasing incidence in hospital-acquired infections, and the adoption of suitable disinfectants is strongly recommended.

Research paper thumbnail of Candidaspecies isolated from different body sites and their antifungal susceptibility pattern: Cross-analysis ofCandida albicansandCandida glabratabiofilms

Medical Mycology, 2016

Candida species are regular commensal in humans, but-especially in immunocompromised patients-the... more Candida species are regular commensal in humans, but-especially in immunocompromised patients-they represent opportunistic pathogens giving rise to systemic infection. The aim of the present work was to isolate and characterize for their antifungal profile Candida species from different body sites and to analyze the biofilms produced by C. albicans and C. glabrata isolates. Eighty-one strains of Candida species from 77 patients were identified. Epidemiological study showed that the most isolated species were C. albicans (44), C. glabrata (13) and C. parapsilosis (13) mainly from Hematology, Infectious Diseases, Medicine, Neonatology and Oncology Divisions, the majority of the biological samples were swabs (44) and blood cultures (16). The analysis of the biofilm formation was performed at 24 and 48-hours comparing resistant and susceptible strains of C. albicans to resistant and susceptible strains of C. glabrata. Candida albicans has a greater ability to form biofilm compared to C. glabrata, both in the susceptible and resistant strains reaching maturity after 24 hours with a complex structure composed of blastospores, pseudohyphae, and hyphae embedded in a matrix. On the contrary, C. glabrata biofilm was composed exclusively of blastospores that in the resistant strain, after 24 hours, were organized in a compact multilayer different to the discontinuous structure observed in the susceptible analyzed strains. In conclusion, the increasing of the incidence of Candida species infection together with their emerging drug resistance also related to the biofilm forming capability underline the need to monitor their distribution and susceptibility patterns for improving the surveillance and for a correct management of the infection.

Research paper thumbnail of In vitroantimicrobial susceptibility ofHelicobacter pylorito nine antibiotics currently used in Central Italy

Scandinavian Journal of Gastroenterology, 2015

Helicobacter pylori expresses an increased resistance in respect to antimicrobials currently used... more Helicobacter pylori expresses an increased resistance in respect to antimicrobials currently used in therapy. The aim of this study was to evaluate the antimicrobial profiles of H. pylori isolates to nine conventional antibiotics used in a Central Region (Abruzzo) of Italy. Biopsies were taken from antrum and fundus of 112 adult and 3 children with Urea Breath Test positive with dyspeptic symptoms and analyzed for H. pylori culture and antibacterial activity. Antimicrobial susceptibility tests were performed for clarithromycin, metronidazole, levofloxacin, moxifloxacin, ciprofloxacin, tetracycline, amoxicillin, ampicillin, and rifabutin by a modified agar dilution susceptibility test. Bacterial culture was successful in 100 out of 115 patients. Helicobacter pylori strains were isolated from 98 antrum and 83 fundus samples. The rate of recovery of H. pylori strains was 90.50% (181/200). The percentages of resistance were as follows: clarithromycin 72.44% antrum, 72.28% fundus; metronidazole 34.69% antrum, 42.16% fundus; levofloxacin 42.85% antrum, 53.01% fundus; moxifloxacin 37.35% antrum, 46.57% fundus; ciprofloxacin 39.47% antrum, 44.28% fundus; tetracycline 2.63% antrum, 2.85% fundus; amoxicillin 1.02% antrum, 1.20% fundus; ampicillin 0% antrum and fundus and rifabutin 0% antrum, 1.20% fundus. A total of 35 subjects harbored multi-resistant strains. This study underlines the high rate of resistance to clarithromycin, metronidazole and quinolones, which may reflect an overuse of them. Culture and susceptibility test, should be performed to prevent the emergence of multi-resistance and to assess an efficacious regimen.

Research paper thumbnail of Carvacrol Codrugs: A New Approach in the Antimicrobial Plan

PloS one, 2015

The increasing prevalence of antibiotic-resistant bacterial infections led to identify alternativ... more The increasing prevalence of antibiotic-resistant bacterial infections led to identify alternative strategies for a novel therapeutic approach. In this study, we synthesized ten carvacrol codrugs - obtained linking the carvacrol hydroxyl group to the carboxyl moiety of sulphur-containing amino acids via an ester bond - to develop novel compounds with improved antimicrobial and antibiofilm activities and reduced toxicity respect to carvacrol alone. All carvacrol codrugs were screened against a representative panel of Gram positive (S. aureus and S. epidermidis), Gram negative (E. coli and P. aeruginosa) bacterial strains and C. albicans, using broth microdilution assays. Results showed that carvacrol codrug 4 possesses the most notable enhancement in the anti-bacterial activity displaying MIC and MBC values equal to 2.5 mg/mL for all bacterial strains, except for P. aeruginosa ATCC 9027 (MIC and MBC values equal to 5 mg/mL and 10 mg/mL, respectively). All carvacrol codrugs 1-10 revea...

Research paper thumbnail of Population dynamics in ageing Helicobacter pylori

The new microbiologica, 2004

The aim of this work was to characterize population changes occurring in aged broth cultures of H... more The aim of this work was to characterize population changes occurring in aged broth cultures of Helicobacter pylori. Experiments were performed using clinical strains cultured immediately after isolation and after multiple subcultures in solid medium. Morphological changes in the ageing bacteria during a 7-day broth culture were analysed by optical and electron microscopy. The expression of the virulence factor, CagA, together with the presence of the cell cycle regulator, cGMP, were also assessed. The transition from bacillary to coccoid forms was the main morphological change observed in freshly isolated bacteria, together with the increase in cGMP from 1 to 2.25 nmoles/mg of proteins within the first 7 days of broth culture. A similar trend of morphological and physiological changes was observed in cells after multiple subcultures in solid medium with a major presence of large cell clusters. The cagA gene product was always expressed in all experimental conditions evaluated. Thes...

Research paper thumbnail of A model of Helicobacter pylori persistence in a case of gastric cancer

The new microbiologica, 2010

The aim of this work was to analyze several clones of Helicobacter pylori isolated from a patient... more The aim of this work was to analyze several clones of Helicobacter pylori isolated from a patient with gastric cancer, to evaluate i) genetic variability ii) virulence factors profile and iii) antimicrobial susceptibility against the drugs commonly used in the H. pylori therapy. A total of 32 H. pylori clones isolated from a biopsy sample coming from a patient with gastric cancer previously treated for H. pylori infection, were analyzed for: the genetic variability by amplified fragment polymorphism analysis; the vacA, cagA virulence status by PCR; the antimicrobial susceptibility by minimum inhibitory concentrations with the agar dilution method towards amoxicillin, clarithromycin, levofloxacin and tinidazole. The patient showed a mixed infection with the presence of at least 3 different strains. The clones isolated possessed the vacA, cagA virulence factors with a different allelic combination (vacA s1/i1/m1; s1/i1i2/m1; s2/i2/m2; s2/i1i2/m2) together with repeated cagA EPIYA moti...

Research paper thumbnail of Central vascular catheter infections in a Hospital of Central Italy

The new microbiologica, 2014

The insertion and the permanence of central venous catheters (CVC) represent potential sources of... more The insertion and the permanence of central venous catheters (CVC) represent potential sources of infection contracted in hospital. The evaluation of the risk of CVC-associated infections was evaluated in a retrospective study during the period 2007-2010 in a Hospital of Central Italy. A total of 514 CVC were collected and examined by microbiological techniques and, among the examined patients, 450 CVC blood cultures were collected. Cultures were performed collecting a portion of 5-6 cm of intravenous catheters in liquid medium and spread on selective media for Gram-positive and Gram-negative bacteria and yeasts; blood specimens were obtained through peripheral venous punctures and analyzed by a commercial automated system. 308/514 (59.90%) samples were positive to the microbiological culture. Staphylococcus aureus, S. epidermidis and other coagulase negative Staphylococci (CNS) were the prevalent Gram-positive bacteria. Among Gram-negative bacteria, Enterobacteriaceae and Pseudomon...

Research paper thumbnail of The effect of a silver nanoparticle polysaccharide system on streptococcal and saliva-derived biofilms

International Journal of Molecular Sciences, 2013

In this work, we studied the antimicrobial properties of a nanocomposite system based on a lactos... more In this work, we studied the antimicrobial properties of a nanocomposite system based on a lactose-substituted chitosan and silver nanoparticles: Chitlac-nAg. Twofold serial dilutions of the colloidal Chitlac-nAg solution were both tested on Streptococcus mitis, Streptococcus mutans, and Streptococcus oralis planktonic phase and biofilm growth mode as well as on saliva samples. The minimum inhibitory and bactericidal concentrations of Chitlac-nAg were evaluated together with its effect on sessile cell viability, as well as both on biofilm formation and on preformed biofilm. In respect to the planktonic bacteria, Chitlac-nAg showed an inhibitory/bactericidal effect against all streptococcal strains at 0.1% (v/v), except for S. mitis ATCC 6249 that was inhibited at one step less. On preformed biofilm, Chitlac-nAg at a value of 0.2%, was able to inhibit the bacterial growth on the supernatant phase as well as on the mature biofilm. For S. mitis ATCC 6249, the biofilm inhibitory concentration of Chitlac-nAg was 0.1%. At sub-inhibitory concentrations, the Streptococcal strains adhesion capability on a polystyrene surface showed a general reduction following a concentration-dependent-way; a similar effect was obtained for the metabolic biofilm activity. From these results, Chitlac-nAg seems to be a promising antibacterial and antibiofilm agent able to hinder plaque formation.

Research paper thumbnail of Dynamic colonizationof Helicobacter pyloriin human gastric mucosa

Scandinavian Journal of Gastroenterology, 2008

To investigate the dynamic behaviour of Helicobacter pylori in the colonization of the human gast... more To investigate the dynamic behaviour of Helicobacter pylori in the colonization of the human gastric mucosa in patients previously treated for H. pylori infection. Twenty-one dyspeptic patients were included in the study. Biopsies from each individual were taken and analysed for H. pylori detection using cultural, molecular and ultrastructural methods. Through culture, H. pylori was isolated in 7 out of 21 patients and the detection of the minimum inhibitory concentration (MIC) against drugs commonly used in H. pylori therapy revealed a susceptibility panel in which only one strain was multidrug resistant. By studying the expression of the H. pylori glmM constitutive gene, viable H. pylori cells were detected in 19 out of 21 analysed biopsies. In these positive cases, the expression of the Quorum-Sensing related gene, luxS, was always detected. The analysis of glmM and luxS sequences confirmed the H. pylori identity. Scanning electron microscopy (SEM) analysis of biopsies from patients harbouring culturable bacteria showed a prevalent "S-shape" H. pylori morphotype co-existent with coccoid aggregated bacteria embedded in an abundant matrix; while samples from patients shown as H. pylori-positive only through the molecular method showed clustered coccoid bacteria arranged in a microbial biofilm. In the present work we describe a new scenario in H. pylori mucosa colonization suggesting, in infection recalcitrance, the planning of more efficacious protocols in order also to identify camouflaged and protected clustered bacteria, taking into account this serious microbial problem in medicine in the recommendation of therapeutic regimens.

Research paper thumbnail of Antibacterial effect of plant extracts againstHelicobacter pylori

Phytotherapy Research, 2005

The aim of this work was to evaluate the antibacterial effect of plant extracts as alternative an... more The aim of this work was to evaluate the antibacterial effect of plant extracts as alternative and[sol ]or as active agents supporting antibiotics for treating Helicobacter pylori infection. The effect of either, ethanolic or aqueous extracts from 17 plant materials were studied against one H. pylori standard strain and 11 clinical isolates using a disc diffusion test and by evaluating the minimum inhibitory concentration (MIC) on solid media. An inhibitory activity against H. pylori strains was recorded in a large percentage of tested plants. MIC values of ethanolic extracts were from two to four concentration steps lower than the aqueous ones. In particular, ethanolic extracts of Cuminum cyminum L. and Propolis expressed MIC90 values of 0.075 mg/mL. The results show a significant in vitro effect of plant extracts against H. pylori that could be considered a valuable support in the treatment of the infection and may contribute to the development of new and safe agents for inclusion in anti-H. pylori regimens.

Research paper thumbnail of Effects of combining extracts (from propolis orZingiber officinale) with clarithromycin onHelicobacter pylori

Phytotherapy Research, 2006

Propolis and Zingiber officinale have been shown to be specifically targeted against Helicobacter... more Propolis and Zingiber officinale have been shown to be specifically targeted against Helicobacter pylori strains, to possess antiinflammatory, antioxidant and antitumoral activity and to be used in traditional medicine for the treatment of gastrointestinal ailments. Considering that these natural products could potentially serve as novel therapeutic tools also in combination with an antibiotic, the aim of this work was to evaluate their effect when combined with clarithromycin on clinical H. pylori isolates (n = 25), characterized in respect to both clarithromycin susceptibility and the presence of the cagA gene. The results showed that the combinations of propolis extract + clarithromycin and Z. officinale extract + clarithromycin exhibited improved inhibition of H. pylori with synergistic or additive activity. Interestingly, the susceptibility to combinations was significantly independent of the microbial clarithromycin susceptibility status. Only one H. pylori strain showed antagonism towards the Z. officinale extract + clarithromycin combination. The data demonstrate that combinations of propolis extract + clarithromycin and Z. officinale extract + clarithromycin have the potential to help control H. pylori-associated gastroduodenal disease.

Research paper thumbnail of Activity of tea tree oil and nerolidol alone or in combination against Pediculus capitis (head lice) and its eggs

Parasitology Research, 2012

Head lice infestation is an emerging social problem in undeveloped and developed countries. Becau... more Head lice infestation is an emerging social problem in undeveloped and developed countries. Because of louse resistance increasing, several long-used insecticidal compounds have lost their efficacy, and alternatives, such as essential oils, have been proposed to treat this parasitic infestation. The present study investigated the efficacy of two natural substances: tea tree (Melaleuca alternifolia) oil and nerolidol (3,7,11-trimethyl-1,6,10-dodecatrien-3-ol) against lice and its eggs. Products were used alone and in combination (ratio 1:1 and 1:2) from 8 % dilution. The in vitro effect of natural substances at different concentrations were evaluated against 69 head lice (adults and nymphs) and 187 louse eggs collected from school children in Chieti-Pescara (Central Italy) over a 6-month period. The lice mortality was evaluated for 24 h by a stereo light microscope. The ovicidal activity was monitored by microscopic inspections for 15 days. Tea tree oil was more effective than nerolidol against head lice with 100 % mortality at 30 min and 1 % concentration. On the contrary, nerolidol expressed a more pronounced ovicidal activity inducing the failure of 50 % of the eggs to hatch at 1 % concentration after 4 days; the same effect was achieved by using a twice concentration of tea tree oil. The association of the two substances both in ratios 1:1 and 1:2 combined efficaciously their insecticidal and ovicidal effect; in particular, the ratio 1:2 (tea tree oil 0.5 % plus nerolidol 1 %) acted producing both the death of all head lice at 30 min and the abortive effect of louse eggs after 5 days. These results offer new potential application of natural compounds and display a promising scenario in the treatment of pediculosis resistant cases. The development of novel pediculicides containing essential oils could be, in fact, an important tool to control the parasitic infestation. E. Di Campli and S. Di Bartolomeo contributed equally to this work.

Research paper thumbnail of Pistacia vera L. oleoresin and levofloxacin is a synergistic combination against resistant Helicobacter pylori strains

Scientific Reports, Mar 15, 2019

The increasing multidrug resistance in Helicobacter pylori, also correlated to its biofilm-formin... more The increasing multidrug resistance in Helicobacter pylori, also correlated to its biofilm-forming ability, underlines the need to search novel strategies to improve the eradication rate. Natural compounds are proposed as antibiotic-resistant-breakers capable to restore the efficacy of conventional drugs. Aim of this work was to evaluate the capability of Pistacia vera L. oleoresin (ORS) to synergize with levofloxacin (LVX) against resistant H. pylori strains. The antimicrobial activity of P. vera L. ORS and LVX and their combinations was determined by MIC/MBC (in neutral and acidic environments) and checkerboard tests. The anti-biofilm effect was determined by biomass quantification. In vivo Galleria mellonella model was used to confirm in vitro data. Pistacia vera L. ORS and LVX MICs ranged respectively from 780 to 3120 mg/l and from 0.12 to 2.00 mg/l, at pH 7.0 and 5.5. MBCs were similar to MICs. Pistacia vera L. ORS was able to synergize with LVX, restoring its effectiveness in LVX resistant strains. Pistacia vera L. ORS, LVX and their synergistic combinations displayed significant biofilm reduction. Pistacia vera L. ORS and LVX, showed protective effect against H. pylori infection on G. mellonella (62% and 63% of survival, respectively). Pistacia vera L. ORS can be considered a promising potentiator to restore the effectiveness of LVX tackling the H. pylori antibiotic resistance phenomenon. Helicobacter pylori is a gastroduodenal pathogen that plays an important role in the pathogenesis of chronic gastritis, peptic ulcer, gastric adenocarcinoma and MALT (mucosa-associated lymphoid tissue) lymphoma. H. pylori infection is difficult to eradicate and requires the combination of different drugs such as clarithromycin, levofloxacin (LVX), amoxicillin, metronidazole, tetracycline and proton pump inhibitor 1-3. The increase of antimicrobial resistance and the failure of therapeutic regimens, strongly underline the need to find novel strategies to enhance the eradication rate, also considering the capability of H. pylori to grow in biofilm mode 4-6. Moreover, the H. pylori antimicrobial resistance profiles vary in different geographic areas, therefore the selection of therapeutic regimens needs to be adjusted according to local resistance patterns, if available 7-10. For all these reasons, the search for alternative and effective new therapeutic schemes is important and urgent 11-14. New approaches to tackle the infections related to multidrug resistant (MDR) bacteria are recently proposed in the search for antibiotic-resistance-breakers capable to synergize with conventional drugs restoring their effectiveness 15-17. To this regards, much interest has been revived in the study of antimicrobial/antivirulence effects of formulations based on medicinal plants 15,18,19. Several plants produce a variety of secondary metabolites such as phenolics, terpenoids and alkaloids, which possess a wide spectrum of biological activities, including the antibacterial ones 14,19-21. They interact with the lipidic bilayer of the cytoplasmic membrane, membrane proteins and enzymes involved in the synthesis of macromolecules, causing increased permeability, loss of proton-motive force and cellular material 14 .

Research paper thumbnail of Microbial Contamination of Smartphone Touchscreens of Italian University Students

Current Microbiology, Dec 15, 2017

In this study, the microbial contamination of smartphones from Italian University students was an... more In this study, the microbial contamination of smartphones from Italian University students was analyzed. A total of 100 smartphones classified as low, medium, and high emission were examined. Bacteria were isolated on elective and selective media and identified by biochemical tests. The mean values of cfu/cm 2 were 0.79 ± 0.01; in particular, a mean of 1.21 ± 0.12, 0.77 ± 0.1 and 0.40 ± 0.10 cfu/cm 2 was present on smartphones at low, medium, and high emission, respectively. The vast majority of identified microorganisms came from human skin, mainly Staphylococci, together with Gram-negative and positive bacilli and yeasts. Moreover, the main isolated species and their mixture were exposed for 3 h to turned on and off smartphones to evaluate the effect of the electromagnetic wave emission on the bacterial cultivability, viability, morphology, and genotypic profile in respect to the unexposed broth cultures. A reduction rate of bacterial growth of 79 and 46% was observed in Staphylococcus aureus and Staphylococcus epidermidis broth cultures, respectively, in the presence of turned on smartphone. No differences in viability were observed in all detected conditions. Small colony variants and some differences in DNA fingerprinting were detected on bacteria when the smartphones were turned on in respect to the other conditions. The colonization of smartphones was limited to human skin microorganisms that can acquire phenotype and genotypic modifications when exposed to microwave emissions.

Research paper thumbnail of Helicobacter pylori Dormant States Are Affected by Vitamin C

International Journal of Molecular Sciences

Helicobacter pylori colonizes human gastric mucosa, overcoming stressful conditions and entering ... more Helicobacter pylori colonizes human gastric mucosa, overcoming stressful conditions and entering in a dormant state. This study evaluated: (i) H. pylori’s physiological changes from active to viable-but-non-culturable (VBNC) and persister (AP) states, establishing times/conditions; (ii) the ability of vitamin C to interfere with dormancy generation/resuscitation. A dormant state was induced in clinical MDR H. pylori 10A/13 by: nutrient starvation (for VBNC generation), incubating in an unenriched medium (Brucella broth) or saline solution (SS), and (for AP generation) treatment with 10xMIC amoxicillin (AMX). The samples were monitored after 24, 48, and 72 h, 8–14 days by OD600, CFUs/mL, Live/Dead staining, and an MTT viability test. Afterwards, vitamin C was added to the H. pylori suspension before/after the generation of dormant states, and monitoring took place at 24, 48, and 72 h. The VBNC state was generated after 8 days in SS, and the AP state in AMX for 48 h. Vitamin C reduced...

Research paper thumbnail of Prebiotic Combinations Effects on the Colonization of Staphylococcal Skin Strains

Microorganisms

Background: An unbalanced skin microbiota due to an increase in pathogenic vs. commensal bacteria... more Background: An unbalanced skin microbiota due to an increase in pathogenic vs. commensal bacteria can be efficiently tackled by using prebiotics. The aim of this work was to identify novel prebiotic combinations by exerting species-specific action between S. aureus and S. epidermidis strains. Methods: First, the antimicrobial/antibiofilm effect of Xylitol-XYL and Galacto-OligoSaccharides–GOS combined with each other at different concentrations (1, 2.5, 5%) against S. aureus and S. epidermidis clinical strains was evaluated in time. Second, the most species-specific concentration was used to combine XYL with Fructo-OligoSaccharides–FOS, IsoMalto-Oligosaccharides–IMO, ArabinoGaLactan–LAG, inulin, dextran. Experiments were performed by OD600 detection, biomass quantification and LIVE/DEAD staining. Results: 1% XYL + 1% GOS showed the best species-specific action with an immediate antibacterial/antibiofilm action against S. aureus strains (up to 34.54% ± 5.35/64.68% ± 4.77) without a re...

Research paper thumbnail of Near-infrared NIR irradiation and sodium hypochlorite: An efficacious association to counteract the Enterococcus faecalis biofilm in endodontic infections

Journal of Photochemistry and Photobiology B: Biology, 2020

, et al., Near-infrared NIR irradiation and sodium hypochlorite: An efficacious association to co... more , et al., Near-infrared NIR irradiation and sodium hypochlorite: An efficacious association to counteract the Enterococcus faecalis biofilm in endodontic infections,

Research paper thumbnail of Graphene Oxide affects Staphylococcus aureus and Pseudomonas aeruginosa dual species biofilm in Lubbock Chronic Wound Biofilm model

Scientific Reports, 2020

Chronic wound management becomes a complex procedure because of the persistence of forming biofil... more Chronic wound management becomes a complex procedure because of the persistence of forming biofilm pathogens that do not respond to antimicrobial treatment. The aim of this paper is to detect the Graphene Oxide-GO effect on Staphylococcus aureus and Pseudomonas aeruginosa dual species wound biofilm in Lubbock Chronic Wound Biofilm-LCWB model. LCWB is a recognized model that mimics the spatial microbial colonization into chronic wounds and reproduces the wound and its clot. Staphylococcus aureus PECHA 10 and P. aeruginosa PECHA 4, are the pathogens used in the study. The GO effect on both in forming and mature biofilms, is detected by the evaluation of the CFU/mg reduction, the cell viability and ultrastructural analysis of the treated LCWBs. Graphene Oxide, at 50 mg/l, shows a significant antibiofilm effect in forming and mature LCWBs. In particular, during the biofilm formation, GO reduces the S. aureus and P. aeruginosa growth of 55.05% ± 4.73 and 44.18% ± 3.91 compared to the con...

Research paper thumbnail of Searching for New Tools to Counteract the Helicobacter pylori Resistance: The Positive Action of Resveratrol Derivatives

Antibiotics, 2020

The drug-resistance phenomenon in Helicobacter pylori underlines the need of novel strategies to ... more The drug-resistance phenomenon in Helicobacter pylori underlines the need of novel strategies to improve the eradication rate including alternative treatments combining antibiotic and non-antibiotic compounds with synergistic action. In this study, the antibacterial (MIC/MBC) and anti-virulence effects (biofilm reduction and swarming motility inhibition) of resveratrol-RSV and new synthetized RSV-phenol derivatives, with a higher bioavailability, alone and combined with levofloxacin-LVX were evaluated against resistant H. pylori clinical strains. The experiments were confirmed in vivo using the Galleria mellonella model. Among the studied RSV derivatives, RSV-3 and RSV-4 possessed higher antibacterial activity with respect to RSV (MICs from 6.25 to 200 µg/mL and from 3.12 to 200 µg/mL, respectively). RSV, RSV-3, and RSV-4 were able to synergize with LVX restoring its effect in two out of seven clinical resistant strains tested for the study. RSV, RSV-3, and RSV-4, alone and with LVX...

Research paper thumbnail of Antimicrobial and Antibiofilm Efficacy of Graphene Oxide against Chronic Wound Microorganisms

Antimicrobial Agents and Chemotherapy, 2018

Chronic wounds represent an increasing problem worldwide. Graphene oxide (GO) has been reported t... more Chronic wounds represent an increasing problem worldwide. Graphene oxide (GO) has been reported to exhibit strong antibacterial activity toward both Gram-positive and Gram-negative bacteria. The aim of this work was to investigate the in vitro antimicrobial and antibiofilm efficacy of GO against wound pathogens. Staphylococcus aureus PECHA 10, Pseudomonas aeruginosa PECHA 4, and Candida albicans X3 clinical isolates were incubated with 50 mg/liter of GO for 2 and 24 h to evaluate the antimicrobial effect. Optical and atomic force microscopy images were performed to visualize the effect of GO on microbial cells. Moreover, the antibiofilm effect of GO was tested on biofilms, both in formation and mature. Compared to the respective time controls, GO significantly reduced the S. aureus growth both at 2 and 24 h in a time-dependent way, and it displayed a bacteriostatic effect in respect to the GO t = 0; an immediate (after 2 h) slowdown of bacterial growth was detected for P. aeruginosa...

Research paper thumbnail of Enterococcus hirae biofilm formation on hospital material surfaces and effect of new biocides

Environmental Health and Preventive Medicine, 2017

Background: Nowadays, the bacterial contamination in the hospital environment is of particular co... more Background: Nowadays, the bacterial contamination in the hospital environment is of particular concern because the hospital-acquired infections (HAIs), also known as nosocomial infections, are responsible for significant morbidity and mortality. This work evaluated the capability of Enterococcus hirae to form biofilm on different surfaces and the action of two biocides on the produced biofilms. Methods: The biofilm formation of E. hirae ATCC 10541 was studied on polystyrene and stainless steel surfaces through the biomass quantification and the cell viability at 20 and 37°C. The effect of LH IDROXI FAST and LH ENZYCLEAN SPRAY biocides on biomasses was expressed as percentage of biofilm reduction. E. hirae at 20 and 37°C produced more biofilm on the stainless steel in respect to the polystyrene surface. The amount of viable cells was greater at 20°C than with 37°C on the two analyzed surfaces. Biocides revealed a good anti-biofilm activity with the most effect for LH ENZYCLEAN SPRAY on polystyrene and stainless steel at 37°C with a maximum biofilm reduction of 85.72 and 86.37%, respectively. Results: E. hirae is a moderate biofilm producer depending on surface material and temperature, and the analyzed biocides express a remarkable antibiofilm action. Conclusion: The capability of E. hirae to form biofilm can be associated with its increasing incidence in hospital-acquired infections, and the adoption of suitable disinfectants is strongly recommended.

Research paper thumbnail of Candidaspecies isolated from different body sites and their antifungal susceptibility pattern: Cross-analysis ofCandida albicansandCandida glabratabiofilms

Medical Mycology, 2016

Candida species are regular commensal in humans, but-especially in immunocompromised patients-the... more Candida species are regular commensal in humans, but-especially in immunocompromised patients-they represent opportunistic pathogens giving rise to systemic infection. The aim of the present work was to isolate and characterize for their antifungal profile Candida species from different body sites and to analyze the biofilms produced by C. albicans and C. glabrata isolates. Eighty-one strains of Candida species from 77 patients were identified. Epidemiological study showed that the most isolated species were C. albicans (44), C. glabrata (13) and C. parapsilosis (13) mainly from Hematology, Infectious Diseases, Medicine, Neonatology and Oncology Divisions, the majority of the biological samples were swabs (44) and blood cultures (16). The analysis of the biofilm formation was performed at 24 and 48-hours comparing resistant and susceptible strains of C. albicans to resistant and susceptible strains of C. glabrata. Candida albicans has a greater ability to form biofilm compared to C. glabrata, both in the susceptible and resistant strains reaching maturity after 24 hours with a complex structure composed of blastospores, pseudohyphae, and hyphae embedded in a matrix. On the contrary, C. glabrata biofilm was composed exclusively of blastospores that in the resistant strain, after 24 hours, were organized in a compact multilayer different to the discontinuous structure observed in the susceptible analyzed strains. In conclusion, the increasing of the incidence of Candida species infection together with their emerging drug resistance also related to the biofilm forming capability underline the need to monitor their distribution and susceptibility patterns for improving the surveillance and for a correct management of the infection.

Research paper thumbnail of In vitroantimicrobial susceptibility ofHelicobacter pylorito nine antibiotics currently used in Central Italy

Scandinavian Journal of Gastroenterology, 2015

Helicobacter pylori expresses an increased resistance in respect to antimicrobials currently used... more Helicobacter pylori expresses an increased resistance in respect to antimicrobials currently used in therapy. The aim of this study was to evaluate the antimicrobial profiles of H. pylori isolates to nine conventional antibiotics used in a Central Region (Abruzzo) of Italy. Biopsies were taken from antrum and fundus of 112 adult and 3 children with Urea Breath Test positive with dyspeptic symptoms and analyzed for H. pylori culture and antibacterial activity. Antimicrobial susceptibility tests were performed for clarithromycin, metronidazole, levofloxacin, moxifloxacin, ciprofloxacin, tetracycline, amoxicillin, ampicillin, and rifabutin by a modified agar dilution susceptibility test. Bacterial culture was successful in 100 out of 115 patients. Helicobacter pylori strains were isolated from 98 antrum and 83 fundus samples. The rate of recovery of H. pylori strains was 90.50% (181/200). The percentages of resistance were as follows: clarithromycin 72.44% antrum, 72.28% fundus; metronidazole 34.69% antrum, 42.16% fundus; levofloxacin 42.85% antrum, 53.01% fundus; moxifloxacin 37.35% antrum, 46.57% fundus; ciprofloxacin 39.47% antrum, 44.28% fundus; tetracycline 2.63% antrum, 2.85% fundus; amoxicillin 1.02% antrum, 1.20% fundus; ampicillin 0% antrum and fundus and rifabutin 0% antrum, 1.20% fundus. A total of 35 subjects harbored multi-resistant strains. This study underlines the high rate of resistance to clarithromycin, metronidazole and quinolones, which may reflect an overuse of them. Culture and susceptibility test, should be performed to prevent the emergence of multi-resistance and to assess an efficacious regimen.

Research paper thumbnail of Carvacrol Codrugs: A New Approach in the Antimicrobial Plan

PloS one, 2015

The increasing prevalence of antibiotic-resistant bacterial infections led to identify alternativ... more The increasing prevalence of antibiotic-resistant bacterial infections led to identify alternative strategies for a novel therapeutic approach. In this study, we synthesized ten carvacrol codrugs - obtained linking the carvacrol hydroxyl group to the carboxyl moiety of sulphur-containing amino acids via an ester bond - to develop novel compounds with improved antimicrobial and antibiofilm activities and reduced toxicity respect to carvacrol alone. All carvacrol codrugs were screened against a representative panel of Gram positive (S. aureus and S. epidermidis), Gram negative (E. coli and P. aeruginosa) bacterial strains and C. albicans, using broth microdilution assays. Results showed that carvacrol codrug 4 possesses the most notable enhancement in the anti-bacterial activity displaying MIC and MBC values equal to 2.5 mg/mL for all bacterial strains, except for P. aeruginosa ATCC 9027 (MIC and MBC values equal to 5 mg/mL and 10 mg/mL, respectively). All carvacrol codrugs 1-10 revea...

Research paper thumbnail of Population dynamics in ageing Helicobacter pylori

The new microbiologica, 2004

The aim of this work was to characterize population changes occurring in aged broth cultures of H... more The aim of this work was to characterize population changes occurring in aged broth cultures of Helicobacter pylori. Experiments were performed using clinical strains cultured immediately after isolation and after multiple subcultures in solid medium. Morphological changes in the ageing bacteria during a 7-day broth culture were analysed by optical and electron microscopy. The expression of the virulence factor, CagA, together with the presence of the cell cycle regulator, cGMP, were also assessed. The transition from bacillary to coccoid forms was the main morphological change observed in freshly isolated bacteria, together with the increase in cGMP from 1 to 2.25 nmoles/mg of proteins within the first 7 days of broth culture. A similar trend of morphological and physiological changes was observed in cells after multiple subcultures in solid medium with a major presence of large cell clusters. The cagA gene product was always expressed in all experimental conditions evaluated. Thes...

Research paper thumbnail of A model of Helicobacter pylori persistence in a case of gastric cancer

The new microbiologica, 2010

The aim of this work was to analyze several clones of Helicobacter pylori isolated from a patient... more The aim of this work was to analyze several clones of Helicobacter pylori isolated from a patient with gastric cancer, to evaluate i) genetic variability ii) virulence factors profile and iii) antimicrobial susceptibility against the drugs commonly used in the H. pylori therapy. A total of 32 H. pylori clones isolated from a biopsy sample coming from a patient with gastric cancer previously treated for H. pylori infection, were analyzed for: the genetic variability by amplified fragment polymorphism analysis; the vacA, cagA virulence status by PCR; the antimicrobial susceptibility by minimum inhibitory concentrations with the agar dilution method towards amoxicillin, clarithromycin, levofloxacin and tinidazole. The patient showed a mixed infection with the presence of at least 3 different strains. The clones isolated possessed the vacA, cagA virulence factors with a different allelic combination (vacA s1/i1/m1; s1/i1i2/m1; s2/i2/m2; s2/i1i2/m2) together with repeated cagA EPIYA moti...

Research paper thumbnail of Central vascular catheter infections in a Hospital of Central Italy

The new microbiologica, 2014

The insertion and the permanence of central venous catheters (CVC) represent potential sources of... more The insertion and the permanence of central venous catheters (CVC) represent potential sources of infection contracted in hospital. The evaluation of the risk of CVC-associated infections was evaluated in a retrospective study during the period 2007-2010 in a Hospital of Central Italy. A total of 514 CVC were collected and examined by microbiological techniques and, among the examined patients, 450 CVC blood cultures were collected. Cultures were performed collecting a portion of 5-6 cm of intravenous catheters in liquid medium and spread on selective media for Gram-positive and Gram-negative bacteria and yeasts; blood specimens were obtained through peripheral venous punctures and analyzed by a commercial automated system. 308/514 (59.90%) samples were positive to the microbiological culture. Staphylococcus aureus, S. epidermidis and other coagulase negative Staphylococci (CNS) were the prevalent Gram-positive bacteria. Among Gram-negative bacteria, Enterobacteriaceae and Pseudomon...

Research paper thumbnail of The effect of a silver nanoparticle polysaccharide system on streptococcal and saliva-derived biofilms

International Journal of Molecular Sciences, 2013

In this work, we studied the antimicrobial properties of a nanocomposite system based on a lactos... more In this work, we studied the antimicrobial properties of a nanocomposite system based on a lactose-substituted chitosan and silver nanoparticles: Chitlac-nAg. Twofold serial dilutions of the colloidal Chitlac-nAg solution were both tested on Streptococcus mitis, Streptococcus mutans, and Streptococcus oralis planktonic phase and biofilm growth mode as well as on saliva samples. The minimum inhibitory and bactericidal concentrations of Chitlac-nAg were evaluated together with its effect on sessile cell viability, as well as both on biofilm formation and on preformed biofilm. In respect to the planktonic bacteria, Chitlac-nAg showed an inhibitory/bactericidal effect against all streptococcal strains at 0.1% (v/v), except for S. mitis ATCC 6249 that was inhibited at one step less. On preformed biofilm, Chitlac-nAg at a value of 0.2%, was able to inhibit the bacterial growth on the supernatant phase as well as on the mature biofilm. For S. mitis ATCC 6249, the biofilm inhibitory concentration of Chitlac-nAg was 0.1%. At sub-inhibitory concentrations, the Streptococcal strains adhesion capability on a polystyrene surface showed a general reduction following a concentration-dependent-way; a similar effect was obtained for the metabolic biofilm activity. From these results, Chitlac-nAg seems to be a promising antibacterial and antibiofilm agent able to hinder plaque formation.

Research paper thumbnail of Dynamic colonizationof Helicobacter pyloriin human gastric mucosa

Scandinavian Journal of Gastroenterology, 2008

To investigate the dynamic behaviour of Helicobacter pylori in the colonization of the human gast... more To investigate the dynamic behaviour of Helicobacter pylori in the colonization of the human gastric mucosa in patients previously treated for H. pylori infection. Twenty-one dyspeptic patients were included in the study. Biopsies from each individual were taken and analysed for H. pylori detection using cultural, molecular and ultrastructural methods. Through culture, H. pylori was isolated in 7 out of 21 patients and the detection of the minimum inhibitory concentration (MIC) against drugs commonly used in H. pylori therapy revealed a susceptibility panel in which only one strain was multidrug resistant. By studying the expression of the H. pylori glmM constitutive gene, viable H. pylori cells were detected in 19 out of 21 analysed biopsies. In these positive cases, the expression of the Quorum-Sensing related gene, luxS, was always detected. The analysis of glmM and luxS sequences confirmed the H. pylori identity. Scanning electron microscopy (SEM) analysis of biopsies from patients harbouring culturable bacteria showed a prevalent "S-shape" H. pylori morphotype co-existent with coccoid aggregated bacteria embedded in an abundant matrix; while samples from patients shown as H. pylori-positive only through the molecular method showed clustered coccoid bacteria arranged in a microbial biofilm. In the present work we describe a new scenario in H. pylori mucosa colonization suggesting, in infection recalcitrance, the planning of more efficacious protocols in order also to identify camouflaged and protected clustered bacteria, taking into account this serious microbial problem in medicine in the recommendation of therapeutic regimens.

Research paper thumbnail of Antibacterial effect of plant extracts againstHelicobacter pylori

Phytotherapy Research, 2005

The aim of this work was to evaluate the antibacterial effect of plant extracts as alternative an... more The aim of this work was to evaluate the antibacterial effect of plant extracts as alternative and[sol ]or as active agents supporting antibiotics for treating Helicobacter pylori infection. The effect of either, ethanolic or aqueous extracts from 17 plant materials were studied against one H. pylori standard strain and 11 clinical isolates using a disc diffusion test and by evaluating the minimum inhibitory concentration (MIC) on solid media. An inhibitory activity against H. pylori strains was recorded in a large percentage of tested plants. MIC values of ethanolic extracts were from two to four concentration steps lower than the aqueous ones. In particular, ethanolic extracts of Cuminum cyminum L. and Propolis expressed MIC90 values of 0.075 mg/mL. The results show a significant in vitro effect of plant extracts against H. pylori that could be considered a valuable support in the treatment of the infection and may contribute to the development of new and safe agents for inclusion in anti-H. pylori regimens.

Research paper thumbnail of Effects of combining extracts (from propolis orZingiber officinale) with clarithromycin onHelicobacter pylori

Phytotherapy Research, 2006

Propolis and Zingiber officinale have been shown to be specifically targeted against Helicobacter... more Propolis and Zingiber officinale have been shown to be specifically targeted against Helicobacter pylori strains, to possess antiinflammatory, antioxidant and antitumoral activity and to be used in traditional medicine for the treatment of gastrointestinal ailments. Considering that these natural products could potentially serve as novel therapeutic tools also in combination with an antibiotic, the aim of this work was to evaluate their effect when combined with clarithromycin on clinical H. pylori isolates (n = 25), characterized in respect to both clarithromycin susceptibility and the presence of the cagA gene. The results showed that the combinations of propolis extract + clarithromycin and Z. officinale extract + clarithromycin exhibited improved inhibition of H. pylori with synergistic or additive activity. Interestingly, the susceptibility to combinations was significantly independent of the microbial clarithromycin susceptibility status. Only one H. pylori strain showed antagonism towards the Z. officinale extract + clarithromycin combination. The data demonstrate that combinations of propolis extract + clarithromycin and Z. officinale extract + clarithromycin have the potential to help control H. pylori-associated gastroduodenal disease.

Research paper thumbnail of Activity of tea tree oil and nerolidol alone or in combination against Pediculus capitis (head lice) and its eggs

Parasitology Research, 2012

Head lice infestation is an emerging social problem in undeveloped and developed countries. Becau... more Head lice infestation is an emerging social problem in undeveloped and developed countries. Because of louse resistance increasing, several long-used insecticidal compounds have lost their efficacy, and alternatives, such as essential oils, have been proposed to treat this parasitic infestation. The present study investigated the efficacy of two natural substances: tea tree (Melaleuca alternifolia) oil and nerolidol (3,7,11-trimethyl-1,6,10-dodecatrien-3-ol) against lice and its eggs. Products were used alone and in combination (ratio 1:1 and 1:2) from 8 % dilution. The in vitro effect of natural substances at different concentrations were evaluated against 69 head lice (adults and nymphs) and 187 louse eggs collected from school children in Chieti-Pescara (Central Italy) over a 6-month period. The lice mortality was evaluated for 24 h by a stereo light microscope. The ovicidal activity was monitored by microscopic inspections for 15 days. Tea tree oil was more effective than nerolidol against head lice with 100 % mortality at 30 min and 1 % concentration. On the contrary, nerolidol expressed a more pronounced ovicidal activity inducing the failure of 50 % of the eggs to hatch at 1 % concentration after 4 days; the same effect was achieved by using a twice concentration of tea tree oil. The association of the two substances both in ratios 1:1 and 1:2 combined efficaciously their insecticidal and ovicidal effect; in particular, the ratio 1:2 (tea tree oil 0.5 % plus nerolidol 1 %) acted producing both the death of all head lice at 30 min and the abortive effect of louse eggs after 5 days. These results offer new potential application of natural compounds and display a promising scenario in the treatment of pediculosis resistant cases. The development of novel pediculicides containing essential oils could be, in fact, an important tool to control the parasitic infestation. E. Di Campli and S. Di Bartolomeo contributed equally to this work.