107 results on '"Alduina, R"'
Search Results
2. Corrigendum to “Soil microbial biomass and bacterial diversity in southern European regions vulnerable to desertification” [Ecol. Indicat. 145 (2022) 109725]
- Author
-
Catania, V., primary, Bueno, R.S., additional, Alduina, R., additional, Grilli, E., additional, La Mantia, T., additional, Castaldi, S., additional, and Quatrini, P., additional
- Published
- 2023
- Full Text
- View/download PDF
3. Soil microbial biomass and bacterial diversity in southern European regions vulnerable to desertification
- Author
-
Catania, V., primary, Bueno, R.S., additional, Alduina, R., additional, Grilli, E., additional, La Mantia, T., additional, Castaldi, S., additional, and Quatrini, P., additional
- Published
- 2022
- Full Text
- View/download PDF
4. Life Desert Adapt, la sfida ambiziosa contro la desertificazione ambientale
- Author
-
Quatrini P., Bueno R. S., Alduina R., Catania V., Cusimano C., Lo Duca R., Lo Verde G., La Mantia T., and Quatrini P., Bueno R. S., Alduina R., Catania V., Cusimano C., Lo Duca R., Lo Verde G., La Mantia T.
- Subjects
Lotta alla desertificazione ,LIFE ,Settore AGR/11 - Entomologia Generale E Applicata ,Settore AGR/05 - Assestamento Forestale E Selvicoltura ,ambienti mediterranei ,diversità ,Settore BIO/19 - Microbiologia Generale ,adattamento dell'agricoltura ai cambiamenti climatici - Abstract
Nel progetto la costruzione di modelli di sviluppo aziendale che adottano tecniche e misure di adattamento capaci di preparare le aree più a rischio.
- Published
- 2018
5. Antibacterial and antitumoral activities of new organotin(IV)-Schiff bases derivatives
- Author
-
Scopelliti, M., Amato, F., Alduina, R., Cancemi, P., Rubino, S., and M. Scopelliti , F. Amato, R. Alduina, P. Cancemi, S. Rubino
- Subjects
Schiff base ,antibacterial ,Organotin ,Settore CHIM/03 - Chimica Generale E Inorganica ,Schiff bases ,antitumor ,Settore BIO/06 - Anatomia Comparata E Citologia ,Settore BIO/19 - Microbiologia Generale - Abstract
This preliminary report shows eight complexes of triorganotin(IV): Ph3SnOH and (CH3)3SnOH with four chelating imines on new synthesis. Of these ligands, two are salen-like (four coordination sites, two imidic, two phenoxidic) [1], one is a tetradentate pyrrole derivative [2] while the fourth, a vita- min B6 derivative, is pentadentate [3]. Ligands have been characterized by means of FT-IR, UV-Vis, Fluorescence, 1H- and 13C-NMR, LC-MS ESI triple quadrupole; complexes by means FT-IR, 1H- and 119Sn-NMR, LC-MS ESI, using the isotopic distribution pattern as a discriminant [4]. Geometry and nature of coordination complexes have been also evaluated using the 119Sn chemical shifts. Solid-state synthesis of the complexes (with a ball mill [5]) was also explored; such method reduces both solvent consumption and time – from 8-10 h under controlled atmosphere to about 1 h, with results identical to the wet synthesis. Antitumoral and antibacterial activities of the triorganotin (IV) complexes (BS01M, BS01P, BS02M, BS02P, BS03M, BS03P, BS04M, BS04P) were tested in vitro. In both analyses, the Shiff bases alone showed no biological activity. Antimicrobial activity was evaluated by Kirby-Bauer method against the Gram-negative Escherichia coli, and the two Gram-positive Kocuria rizophila and Staphylococcus aureus strains. All the ML2 complexes were active in inhibiting bacterial growth, with BS02P and BS03P showing the best antibacterial performance. Among the ML complexes, BS01M was not active, BS02M showed a weak antibacterial activity only against the Gram-positive bacteria, BS04M was mainly active against the Gram-negative E. coli and BS03M was active against all the tested strains. Antitumor activity was evaluated by MTT assay against cervical (HeLa), colon adenocarcinoma (HT- 29) and breast (MDA-MB231) cancer cell lines. Results showed that ML2 complexes are more active than ML ones, with HeLa cells more sensitive to treatments. These complexes (especially the ML2) showed promising results; their mechanism of action is under investigation. References 1. K. Tayade, S.K. Sahoo, S. Chopra, N. Singh, Inorganica Chimica Acta, 421, 538-543 (2014); https://doi.org/10.1016/j. ica.2014.05.014 2. S. Meghdadi, M. Amirnasr, K. Mereiter, Polyhedron, 30, 1651-1656 (2011); https://doi.org/10.1016/j.poly.2011.03.041 3. D. Sharma, S.K. Sahoo, S. Chaudhary, R. Kanta Bera, J.F. Callan, Analyst, 138, 3646-3650 (2013); https://doi.org/10.1039/ C3AN00199G 4. G. Lawson, R.H. Dahm, N. Ostah, E.D. Woodland, Applied Organometallic Chemistry, 10, 125-133 (1996); https://doi. org/10.1002/(SICI)1099-0739(199603)10:23.0.CO;2-1 5. G.A. Bowmaker, Chemical Communications, 49, 334-348 (2013); http://doi.org/10.1039/C2CC35694E 6. R. Di Stefano, M. Scopelliti, C. Pellerito, G. Casella, T. Fiore, G.C. Stocco, R. Vitturi, L. Ronconi, I.D. Sciacca, L. Pellerito, Journal of Inorganic Biochemistry, 98, 534-546 (2004); https://doi.org/10.1016/j.jinorgbio.2003.12.013 7. C. Pellerito, L. Nagy, L. Pellerito, A. Szorcsik, Journal of Organometallic Chemistry, 691, 1733-1747 (2006); https://doi. org/10.1016/j.jorganchem.2005.12.025
- Published
- 2018
6. Molecular systematics of two Mediterranean blenniid species from the genus Salaria (Perciformes, Blenniidae): Salaria basilisca and S. pavo
- Author
-
Belaiba, E., Marrone, F., Alduina, R., Bahri-Sfar, L., Arculeo, M., Belaiba, E, Marrone, F, Alduina, R, Bahri-Sfar, L, and Arculeo, M
- Subjects
Salaria ,Molecular Systematics ,Settore BIO/05 - Zoologia ,Blenniidae - Published
- 2016
7. Synthesis, properties, antitumor and antibacterial activity of new Pt(II) and Pd(II) complexes with 2,20-dithiobis(benzothiazole) ligand
- Author
-
Rubino, S., Busà, R., Attanzio, A., Alduina, R., Di Stefano, V., Girasolo, M., Orecchio, S., and Tesoriere, L.
- Subjects
Platinum complex Palladium complex Heterocyclic nitrogen ligand Anticancer activity Antimicrobial activity ,Settore CHIM/03 - Chimica Generale E Inorganica - Published
- 2017
8. Guanosine 5'-diphosphate 3'-diphosphate (ppGpp) as a negative modulator of polynucleotide phosphorylase activity in a 'rare' actinomycete
- Author
-
SICULELLA, Luisa, DAMIANO, FABRIZIO, GNONI, Gabriele Vincenzo, ALIFANO, Pietro, di Summa R, Tredici SM, Alduina R, Siculella, L, Damiano, F, di Summa, R, Tredici, SM, Alduina, R, Gnoni, GV, Alifano, P, Siculella, Luisa, Damiano, Fabrizio, Tredici, Sm, Gnoni, Gabriele Vincenzo, and Alifano, Pietro
- Subjects
ppGpp ,Polynucleotide phosphorylase ,Actinomycete ,ppGpp, Actinomycetes - Abstract
With the beginning of the idiophase the highly phosphorylated guanylic nucleotides guanosine 5'-diphosphate 3'-diphosphate (ppGpp) and guanosine 5'-triphosphate 3'-diphosphate (pppGpp), collectively referred to as (p)ppGpp, activate stress survival adaptation programmes and trigger secondary metabolism in actinomycetes. The major target of (p)ppGpp is the RNA polymerase, where it binds altering the enzyme activity. In this study analysis of the polynucleotide phosphorylase (PNPase)-encoding gene pnp mRNA, in Nonomuraea sp. ATCC 39727 wild-type, constitutively stringent and relaxed strains, led us to hypothesize that in actinomycetes (p)ppGpp may modulate gene expression at the level of RNA decay also. This hypothesis was supported by: (i) in vitro evidence that ppGpp, at physiological levels, inhibited both polynucleotide polymerase and phosphorolytic activities of PNPase in Nonomuraea sp., but not in Escherichia coli, (ii) in vivo data showing that the pnp mRNA and the A40926 antibiotic cluster-specific dpgA mRNA were stabilized during the idiophase in the wild-type strain but not in a relaxed mutant and (iii) measurement of chemical decay of pulse-labelled bulk mRNA. The results of biochemical tests suggest competitive inhibition of ppGpp with respect to nucleoside diphosphates in polynucleotide polymerase assays and mixed inhibition with respect to inorganic phosphate when the RNA phosphorolytic activity was determined.
- Published
- 2010
9. Epigenetic influence on Streptomyces coelicolor morphological and physiological differentiation
- Author
-
Annalisa, P., Federica, A., Angel, M., and Alduina, R.
- Subjects
Streptomyces coelicolor, epigenetics, morphological and physiological differentiation - Abstract
DNA methylation is an epigenetic modification regulating many aspects of biological processes; for instance, in bacteria adenine methylation is well known to be associated with DNA repair and coordination of replication, while the role of cytosine methylation has been elucidated only in a few examples. Streptomyces coelicolor is a mycelial soil microorganism, producer of several antibiotics, with a complex life cycle that includes three different cell types: unigenomic spores, a compartmentalized mycelium (MI) and a multinucleated mycelium (MII). The main objective of this study was to investigate the role of DNA cytosine methylation along the morphological and physiological differentiation of S. coelicolor. Liquid and solid cultures of S. coelicolor were treated with 5-aza-2'-deoxycytidine (aza-dC, a cytidine analogous that inhibits DNA-methyltransferase activity) demonstrating that methylation influences spore germination, aerial mycelium formation and antibiotic production. Sporulation on solid culture was also affected. To further demonstrate the role of cytosine methylation, knock out mutants in putative methyltransferase genes were generated. Interestingly, disruption of a methyltransferase gene caused a delay in physiological and morphological differentiation similarly to what was obtained treating the cultures with aza-dC. Altogether these results demonstrate the involvement of DNA cytosine methylation on both morphological and physiological differentiation of S. coelicolor.
10. Production of Antibacterial Compounds from Actinomycetes
- Author
-
Rosa Alduina, Letizia Lo Grasso, Delia Francesca Chillura Martino, Dharumadurai Dhanasekaran e Yi Jiang, Lo Grasso, L, Chillura Martino, D, Alduina, R, Lo Grasso, L., Chillura-Martino, D., and Alduina, R.
- Subjects
Production (economics) ,Food science ,Biology ,Settore BIO/19 - Microbiologia Generale ,Antibiotic production, Actinomycetes, genetic engineering, regulation, heterologous expression - Abstract
Actinomycetes are soil-dwelling Gram-positive bacteria, industrially relevant as producers of a wide range of bioactive secondary metabolites, including many antibiotics of clinical and commercial importance.The understanding of actinomycete biology has been based on extensive studies on the model organism Streptomyces coelicolor over many years and on the availability of its complete genome sequence. This bacterium has an unusual complex developmental cycle that includes programmed cell death phenomena that make this bacterium a multicellular prokaryotic model.Morphological differentiation in S. coelicolor is strictly related to physiological differentiation: indeed the onset of morphological differentiation generally coincides with the production of secondary metabolites. During cell death, degradative proteins are synthesized and involved in an extensive degradation of some cellular constituents (proteins and lipids) used for a second growth phase, that is accompanied by antibiotic production.If on one hand, many factors with pleiotropic activity have been identified as key players to control both morphological and physiological differentiation in S. coelicolor, on the other hand, for most actinomycetes, mechanisms and factors governing morphological and physiological processes have not been deeply investigated.This chapter reviews the regulatory mechanisms known to control antibiotic production in actinomycetes and both genetic and physiological methods adopted to improve antibiotic yields.
- Published
- 2016
11. Pulsed field gel electrophoresis and genome size estimates
- Author
-
Rosa Alduina, Annalisa Pisciotta, Alduina, R, Pisciotta, A, Alduina, R., and Pisciotta, A.
- Subjects
Genome size ,DNA, Bacterial ,Materials science ,Chromosomes, Archaeal ,Settore BIO/19 - Microbiologia Generale ,Genome ,Plasmid ,chemistry.chemical_compound ,Genetic ,Genome, Archaeal ,Electric field ,Pulsed-field gel electrophoresis ,Molecular Biology ,Electrophoresis, Agar Gel ,Base Composition ,Strain (chemistry) ,Bacteria ,Multi-replicon ,Medicine (all) ,Physical Chromosome Mapping ,food and beverages ,Chromosomes, Bacterial ,Archaea ,Electrophoresis, Gel, Pulsed-Field ,DNA, Archaeal ,chemistry ,Megaplasmid ,Biological system ,DNA ,Genome, Bacterial ,Genome topology - Abstract
Pulsed field gel electrophoresis (PFGE) is a quick and reliable procedure to resolve DNA molecules larger than 30 kb by applying an electric field that periodically changes direction. This technique can be used to estimate genome size of a microorganism, to reveal if a genome is circular or linear, to indicate the presence of megaplasmids, and to show if a strain contains only one or more chromosomes.
- Published
- 2015
12. Tolerance, Adaptation, and Cell Response Elicited by Micromonospora sp. Facing Tellurite Toxicity: A Biological and Physical-Chemical Characterization
- Author
-
Francesco Carfì Pavia, Simona Campora, DAVIDE ZANNONI, Delia Chillura Martino, Raymond J. Turner, Alessandro Presentato, RValeria Alduina, Elena Piacenza, Vito Armando Laudicina, Piacenza E., Campora S., Carfì Pavia F., Chillura Martino D.F., Laudicina V.A., Alduina R., Turner R.J., Zannoni D., and Presentato A.
- Subjects
multivariate statistical analysis ,Extracellular Polymeric Substance Matrix ,Superoxide Dismutase ,Organic Chemistry ,General Medicine ,Settore BIO/19 - Microbiologia Generale ,fatty acids ,Micromonospora ,Catalysis ,Computer Science Applications ,Inorganic Chemistry ,FTIR spectroscopy ,Protein Aggregates ,tellurite ,bacterial cell membrane ,cell morphology changes ,oxidative stress ,heavy metals ,Physical and Theoretical Chemistry ,Tellurium ,Molecular Biology ,Spectroscopy - Abstract
The intense use of tellurium (Te) in industrial applications, along with the improper disposal of Te-derivatives, is causing their accumulation in the environment, where oxyanion tellurite (TeO32−) is the most soluble, bioavailable, and toxic Te-species. On the other hand, tellurium is a rare metalloid element whose natural supply will end shortly with possible economic and technological effects. Thus, Te-containing waste represents the source from which Te should be recycled and recovered. Among the explored strategies, the microbial TeO32− biotransformation into less toxic Te-species is the most appropriate concerning the circular economy. Actinomycetes are ideal candidates in environmental biotechnology. However, their exploration in TeO32− biotransformation is scarce due to limited knowledge regarding oxyanion microbial processing. Here, this gap was filled by investigating the cell tolerance, adaptation, and response to TeO32− of a Micromonospora strain isolated from a metal(loid)-rich environment. To this aim, an integrated biological, physical-chemical, and statistical approach combining physiological and biochemical assays with confocal or scanning electron (SEM) microscopy and Fourier-transform infrared spectroscopy in attenuated total reflectance mode (ATR-FTIR) was designed. Micromonospora cells exposed to TeO32− under different physiological states revealed a series of striking cell responses, such as cell morphology changes, extracellular polymeric substance production, cell membrane damages and modifications, oxidative stress burst, protein aggregation and phosphorylation, and superoxide dismutase induction. These results highlight this Micromonospora strain as an asset for biotechnological purposes.
- Published
- 2022
13. Cross-linked natural IntegroPectin films from citrus biowaste with intrinsic antimicrobial activity
- Author
-
Elena Piacenza, Alessandro Presentato, Rosa Alduina, Antonino Scurria, Mario Pagliaro, Lorenzo Albanese, Francesco Meneguzzo, Rosaria Ciriminna, Delia F. Chillura Martino, Piacenza E., Presentato A., Alduina R., Scurria A., Pagliaro M., Albanese L., Meneguzzo F., Ciriminna R., and Chillura Martino D.F.
- Subjects
Polymers and Plastics ,Antimicrobial films ,Terpenes ,Controlled release ,Polyphenols ,Settore BIO/19 - Microbiologia Generale ,Citrus pectin ,Pectin films ,Settore CHIM/02 - Chimica Fisica - Abstract
Pectin recovered via hydrodynamic cavitation (IntegroPectin) from lemon and grapefruit agri-food waste intrinsically containing antimicrobial bioactive substances (flavonoids, phenolic acids, terpenes, and terpenoids) was used to generate innovative and eco-compatible films that efficiently inhibit the growth of Gram-negative pathogens. Extensive characterization of films confirmed the presence of these substances, which differently interact with the polysaccharide polymer (pectin), plasticizer (glycerol), surfactant (Tween 60), and cross-linker (Ca2+), conferring to these films a unique structure. Besides, IntegroPectin-based films constitute versatile systems for the sustained, controlled, and slow-release (up to 72 h) of bioactive substances in an aqueous environment. This feature is crucial for the good in vitro antimicrobial activity exerted by IntegroPectin films against three Gram-negative bacteria (two indicator pathogen strains Pseudomonas aeruginosa ATCC 10145, P. aeruginosa PAO1, and the clinical isolate Klebsiella pneumoniae) that are involved in the global emergence of the antimicrobial resistance. Graphical abstract
- Published
- 2022
14. Unveiling the egg microbiota of the loggerhead sea turtle Caretta caretta in nesting beaches of the Mediterranean Sea
- Author
-
Luca Vecchioni, Antonino Pace, Arianna Sucato, Flavia Berlinghieri, Irene Cambera, Giulia Visconti, Sandra Hochscheid, Marco Arculeo, Rosa Alduina, Groothuis lab, Vecchioni L., Pace A., Sucato A., Berlinghieri F., Cambera I., Visconti G., Hochscheid S., Arculeo M., and Alduina R.
- Subjects
Bird eggs, turtles, eggs, gut bacteria, beaches, actinodacteria, bacteria, gastrointestinal tract ,Multidisciplinary ,Bacteria ,Sand ,caretta caretta ,RNA, Ribosomal, 16S ,microbiota ,Mediterranean Sea ,Animals ,egg ,human activities ,Nesting Behavior ,Turtles - Abstract
Microbes have central roles in the development and health of animals, being the introduction of specific microbial species a potential conservation strategy to protect animals from emerging diseases. Thus, insight into the microbiota of the species and their habitats is essential. In this manuscript, we report for the first time the bacterial composition of all the components (eggshells of hatched and unhatched eggs, internal content of unhatched eggs, intestinal content of hatchling and pipping sea turtles, and sand) of three nesting beaches of Caretta caretta along the Italian coasts of the Mediterranean Sea. The analysis of 26 amplicon samples was carried out using next-generation sequencing analysis, targeting V3–V4 regions of the bacterial 16S rRNA gene. Samples featured mainly Proteobacteria, Actinobacteria, Firmicutes, and Bacteroidetes, whose percentages depended on the sample type. Our results showed that, although from different sampling sites, the internal content of the unhatched eggs, intestinal content of hatchling and pipping sea turtles share the microbiota, which was yet different from that of eggshells and sand of the same nesting beach. This study suggests the maternal and environmental influence alongside a protective role of eggshells in shaping the egg microbiota of Caretta caretta sea turtles.
- Published
- 2022
15. Occurrence of Antibiotic Resistance in the Mediterranean Sea
- Author
-
Delia Gambino, Dario Savoca, Arianna Sucato, Valeria Gargano, Antonino Gentile, Licia Pantano, Domenico Vicari, Rosa Alduina, Gambino D., Savoca D., Sucato A., Gargano V., Gentile A., Pantano L., Vicari D., and Alduina R.
- Subjects
Microbiology (medical) ,Bla ,antibiotic-resistant bacteria ,antibiotic resistance genes ,int1 ,blaTEM ,sulII ,heavy metal resistance genes ,Biochemistry ,Microbiology ,Antibiotic-resistant bacteria ,Heavy metal resistance genes ,Infectious Diseases ,Antibiotic resistance genes ,TEM ,SulII ,Pharmacology (medical) ,General Pharmacology, Toxicology and Pharmaceutics ,Int1 - Abstract
Seawater could be considered a reservoir of antibiotic-resistant bacteria and antibiotic resistance genes. In this communication, we evaluated the presence of bacterial strains in seawater collected from different coasts of Sicily by combining microbiological and molecular methods. Specifically, we isolated viable bacteria that were tested for their antibiotic resistance profile and detected both antibiotic and heavy metal resistance genes. Both antibiotic-resistant Gram-negative bacteria, Vibrio and Aeromonas, and specific antibiotic resistance genes were found in the seawater samples. Alarming levels of resistance were determined towards cefazolin, streptomycin, amoxicillin/clavulanic acid, ceftriaxone, and sulfamethoxazole/trimethoprim, and mainly genes conferring resistance to β-lactamic and sulfonamide antibiotics were detected. This survey, on the one hand, presents a picture of the actual situation, showing the pollution status of the Tyrrhenian coast of Sicily, and, on the other hand, can be considered as a baseline to be used as a reference time for future analysis.
- Published
- 2022
- Full Text
- View/download PDF
16. Water Resource Recovery Facilities (WRRFs): The Case Study of Palermo University (Italy)
- Author
-
Giorgio Mannina, Sofia Maria Muscarella, Alida Cosenza, Vito Armando Laudicina, Dario Presti, Rosa Alduina, Luigi Badalucco, Daniele Di Trapani, Giuseppe Gallo, Lorenzo Barbara, Claire Capri, Mannina G., Alduina R., Badalucco L., Barbara L., Capri F.C., Cosenza A., Di Trapani D., Gallo G., Laudicina V.A., Muscarella S.M., and Presti D.
- Subjects
Waste management ,Water supply for domestic and industrial purposes ,Circular economy ,Geography, Planning and Development ,Hydraulic engineering ,Wastewater treatment ,Aquatic Science ,Biochemistry ,Volatile fatty acids ,Wastewater ,Water smart solutions ,Sustainability ,Environmental science ,Sewage treatment ,Batch test ,circular economy ,wastewater treatment ,water resource ,water smart solutions ,TC1-978 ,TD201-500 ,Water resource ,Water Science and Technology ,Resource recovery - Abstract
The wastewater sector paradigm is shifting from wastewater treatment to resource recovery. In addition, concerns regarding sustainability during the operation have increased. In this sense, there is a need to break barriers (i.e., social, economic, technological, legal, etc.) for moving forward towards water resource recovery facilities and demonstration case studies can be very effective and insightful. This paper presents a new water resource recovery case study which is part of the Horizon 2020 EU Project “Achieving wider uptake of water-smart solutions—Wider Uptake”. The final aim is to demonstrate the importance of a resource recovery system based on the circular economy concept. The recovery facilities at Palermo University (Italy) are first presented. Afterwards, the resource recovery pilot plants are described. Preliminary results have underlined the great potential of the wastewater treatment plant in terms of resources recovery and the central role of the University in fostering the transition towards circular economy. The fermentation batch test highlighted a volatile fatty acids (VFAs) accumulation suitable for polyhydroxyalkanoates (PHAs) production. The results of static adsorption and desorption tests showed that the highest amount of adsorbed NH4+ was recorded for untreated and HCl-Na treated clinoptilolite.
- Published
- 2021
- Full Text
- View/download PDF
17. Conservation state of two paintings in the Santa Margherita cliff cave: role of the environment and of the microbial community
- Author
-
Francesco Armetta, Josue Cardenas, Rosa Alduina, Luca Vecchioni, Alessandro Presentato, Maria Luisa Saladino, Eugenio Caponetti, Alberto Spinella, Pietro Di Stefano, Armetta F., Cardenas J., Caponetti E., Alduina R., Presentato A., Vecchioni L., di Stefano P., Spinella A., and Saladino M.L.
- Subjects
Health, Toxicology and Mutagenesis ,media_common.quotation_subject ,Natural (archaeology) ,Cave painting ,State (polity) ,Cave ,Cliff ,Environmental Chemistry ,Sulfates and chlorides ,media_common ,geography ,Painting ,geography.geographical_feature_category ,Bacteria ,Microbiota ,Temperature ,Conservation Plan ,Plan (archaeology) ,Humidity ,Environmental monitoring ,General Medicine ,Pollution ,Archaeology ,humanities ,Conservation plan ,Paintings ,Bacterial community - Abstract
The conservation of ancient paintings sited in humid environments is an actual challenge for restorers, because it needs the knowledge of the materials the paintings are made up and of their interaction with a peculiar surrounding environment; thus, tailored procedures and strategies aimed at restoring and preserving paintings are necessary. The Santa Margherita’s cave in Castellammare del Golfo (Trapani, Italy) is a natural cave, containing the remains of paintings, in a poor state of conservation, belonging to an ancient church dated back to the Middle Age. The present manuscript reports the monitoring of environmental conditions (i.e., temperature and humidity) in a full year as well as a study on the materials constituting the stone support and the paintings together with a survey of the microbial community. The findings allow us to define the causes which mainly involve the degradation of the paintings. In detail, the degradation of the east and the west wall occurred differently because of the exposure to the sea aerosol, which influenced the salt composition, also contributing to diversifying the bacterial community. Some specific actions to plan the conservation and restoration of paintings and to preserve the site are suggested.
- Published
- 2021
18. Influence of volatile solids and pH for the production of volatile fatty acids: batch fermentation tests using sewage sludge
- Author
-
Fanny Claire Capri, Rosa Alduina, Giorgio Mannina, Alida Cosenza, Giuseppe Gallo, Dario Presti, Presti D., Cosenza A., Capri F.C., Gallo G., Alduina R., and Mannina G.
- Subjects
Environmental Engineering ,Circular economy ,Batch fermentation ,0207 environmental engineering ,Bioengineering ,02 engineering and technology ,010501 environmental sciences ,01 natural sciences ,Bioreactors ,Nutrient ,Volatile fatty acids ,Resource recovery from wastewater ,020701 environmental engineering ,Waste Management and Disposal ,0105 earth and related environmental sciences ,Suspended solids ,Settore ICAR/03 - Ingegneria Sanitaria-Ambientale ,Bacteria ,Sewage ,Renewable Energy, Sustainability and the Environment ,Chemistry ,General Medicine ,Hydrogen-Ion Concentration ,Other Quantitative Biology (q-bio.OT) ,Fatty Acids, Volatile ,Pulp and paper industry ,6. Clean water ,Quantitative Biology - Other Quantitative Biology ,Activated sludge ,FOS: Biological sciences ,Fermentation ,Bacterial community ,Retention time ,Sludge - Abstract
The aim of this work was to study the effect of volatile suspended solid (VSS) and pH on volatile fatty acids (VFA) production from waste activated sludge (WAS) fermentation by means of batch tests. The final goal was to gain insights to enhance VFA stream quality, with the novelty of using WAS with high sludge retention time. Results revealed that the optimum conditions to maximize VFAs and minimize nutrients and non-VFA sCOD are a VSS concentration of 5.9 g/L and initial pH adjustment to pH 10. The WAS bacterial community structures were analysed according to Next Generation Sequencing (NGS) of 16S rDNA amplicons. The results revealed changes of bacterial phyla abundance in comparison with the batch test starting condition., 27 pages, 4 figures
- Published
- 2021
19. Comparison of the intestinal microbiome of italian patients with multiple sclerosis and their household relatives
- Author
-
Domenico Nuzzo, Luca Scalisi, Fanny Claire Capri, Sabrina Realmuto, Luca Vecchioni, Salvatore Cottone, Rosa Alduina, Paola Galluzzo, Galluzzo P., Capri F.C., Vecchioni L., Realmuto S., Scalisi L., Cottone S., Nuzzo D., and Alduina R.
- Subjects
Ruminococcaceae ,Bacteroidaceae ,Science ,Veillonellaceae ,Disease ,Gut flora ,General Biochemistry, Genetics and Molecular Biology ,Article ,Multiple sclerosis ,medicine ,Microbiome ,Desulfovibrionaceae christensenellaceae ,Ecology, Evolution, Behavior and Systematics ,Genetics ,Clostridiales ,biology ,Burkholderiaceae ,Paleontology ,biology.organism_classification ,medicine.disease ,Space and Planetary Science ,Tannerellaceae - Abstract
Multiple sclerosis (MS) is a chronic immune-mediated disease of the central nervous system, caused by a combination of genetic and environmental factors. In recent years, a role in MS pathogenesis was assigned to the gut microbiota. However, different signatures of gut dysbiosis have been shown to depend on environmental factors, like diet and lifestyle. In this study, we compared the gut microbiome in MS patients and their household healthy relatives sharing lifestyle and environmental factors. Faecal metagenomic DNA was extracted and the V3–V4 regions of the conserved bacterial 16S ribosomal RNA gene were amplified and sequenced. While overall bacterial communities were similar, specific families differed between healthy and MS subjects. We observed an increase in Ruminococcaceae, Christensenellaceae, Desulfovibrionaceae, Clostridiales, and Family XIII in MS patients, while Bacteroidaceae, Tannerellaceae, Veillonellaceae, and Burkholderiaceae were more abundant in healthy controls. In addition, principle coordinate analysis showed that the gut microbiome of all MS patients formed a cluster being less diverse than the household relatives and that gut microbiota of MS patients with EDSS 4.5–7 formed a distinct cluster in respect to their controls. Overall, our study is consistent with the hypothesis that MS patients have gut microbial dysbiosis and evidenced the importance of environmental factors in shaping the gut microbiome.
- Published
- 2021
20. New neuroprotective effect of lemon integropectin on neuronal cellular model
- Author
-
Costanza Giardina, Valentina Di Liberto, Miriana Scordino, Mario Pagliaro, Pasquale Massimo Picone, Domenico Nuzzo, Francesco Meneguzzo, Rosa Alduina, Rosaria Ciriminna, Antonino Scurria, Giuseppa Mudò, Alessandro Presentato, Laura M. Ilharco, Alexandra Fidalgo, Nuzzo D., Picone P., Giardina C., Scordino M., Mudo' G., Pagliaro M., Scurria A., Meneguzzo F., Ilharco L.M., Fidalgo A., Alduina R., Presentato A., Ciriminna R., and Di Liberto V.
- Subjects
Cell signaling ,antioxidant ,Physiology ,hesperidin ,Antioxidant, Flavonoids, Hesperidin, Mitochondria, Neu-roprotective, Neurological disease, Oxidative stress, Pectin ,Clinical Biochemistry ,RM1-950 ,Mitochondrion ,Cell morphology ,medicine.disease_cause ,Biochemistry ,Neuroprotection ,Article ,flavonoids ,03 medical and health sciences ,0302 clinical medicine ,medicine ,oxidative stress ,Viability assay ,neurological disease ,Molecular Biology ,030304 developmental biology ,chemistry.chemical_classification ,pectin ,neuroprotective ,0303 health sciences ,Reactive oxygen species ,Cell Biology ,Cell biology ,mitochondria ,chemistry ,Therapeutics. Pharmacology ,Cellular model ,030217 neurology & neurosurgery ,Oxidative stress - Abstract
Lemon IntegroPectin obtained via hydrodynamic cavitation of organic lemon processing waste in water shows significant neuroprotective activity in vitro, as first reported in this study investigating the effects of both lemon IntegroPectin and commercial citrus pectin on cell viability, cell morphology, reactive oxygen species (ROS) production, and mitochondria perturbation induced by treatment of neuronal SH-SY5Y human cells with H2O2. Mediated by ROS, including H2O2 and its derivatives, oxidative stress alters numerous cellular processes, such as mitochondrial regulation and cell signaling, propagating cellular injury that leads to incurable neurodegenerative diseases. These results, and the absence of toxicity of this new pectic substance rich in adsorbed flavonoids and terpenes, suggest further studies to investigate its activity in preventing, retarding, or even curing neurological diseases.
- Published
- 2021
21. Flavonoids in Lemon and Grapefruit IntegroPectin
- Author
-
Rosaria Ciriminna, Domenico Nuzzo, Antonino Scurria, Rosa Alduina, Mario Pagliaro, Federica Zabini, Alessandro Presentato, Marzia Sciortino, Lorenzo Albanese, Giuseppe Avellone, Francesco Meneguzzo, Scurria A., Sciortino M., Albanese L., Nuzzo D., Zabini F., Meneguzzo F., Alduina R., Presentato A., Pagliaro M., Avellone G., and Ciriminna R.
- Subjects
Citrus ,food.ingredient ,Pectin ,Flavonoid ,citrus fruits ,Terpene ,pectins ,chemistry.chemical_compound ,Hesperidin ,food ,hydrodynamic cavitation ,Food science ,QD1-999 ,Naringin ,IntegroPectin ,Flavonoids ,chemistry.chemical_classification ,Full Paper ,food and beverages ,Neurodegenerative Diseases ,General Chemistry ,Full Papers ,food_chemistry ,Chemistry ,chemistry ,Fruit ,Eriocitrin ,Kaempferol ,Citrus paradisi - Abstract
Following the analysis of terpenes present in new lemon and grapefruit “IntegroPectin” pectins obtained via the hydrodynamic cavitation of industrial lemon and grapefruit processing waste, the HPLC‐MS analysis of flavonoid and other phenolic compounds reveals the presence of eriocitrin, naringin, hesperidin and kaempferol typical of the respective citrus fruits. The pectic fibers rich in rhamnogalacturonan‐I regions act as chemical sponges adsorbing and concentrating at their outer surface highly bioactive citrus flavonoids and terpenes. These findings, together with the unique molecular structure of these new whole citrus pectins, provide preliminary insight into the broad‐scope biological activity of these new biomaterials. Numerous new biomedical applications are anticipated, including likely use in the prevention and treatment of microbial infections and neurodegenerative disease., Molecular synergy in action, IntegroPectin! The pectic fibers rich in rhamnogalacturonan‐I regions of lemon and grapefruit “IntegroPectin” pectins obtained via the hydrodynamic cavitation of industrial lemon and grapefruit processing waste, adsorb and concentrate at their outer surface highly bioactive citrus flavonoids and terpenes. Flavonoid compounds include eriocitrin, naringin, hesperidin and kaempferol typical of the respective citrus fruits.
- Published
- 2021
22. Volatile Compounds of Lemon and Grapefruit IntegroPectin
- Author
-
Alessandro Presentato, Delia Francesca Chillura Martino, Claudia Lino, Antonino Scurria, Domenico Nuzzo, Elena Piacenza, Lorenzo Albanese, Francesco Meneguzzo, Rosaria Ciriminna, Rosa Alduina, Mario Pagliaro, Giuseppe Avellone, Marzia Sciortino, Federica Zabini, Scurria A., Sciortino M., Presentato A., Lino C., Piacenza E., Albanese L., Zabini F., Meneguzzo F., Nuzzo D., Pagliaro M., Chillura Martino D.F., Alduina R., Avellone G., and Ciriminna R.
- Subjects
Citrus ,food.ingredient ,Pectin ,lemon ,Phytochemicals ,Pharmaceutical Science ,grapefruit ,Article ,Industrial waste ,Gas Chromatography-Mass Spectrometry ,Analytical Chemistry ,lcsh:QD241-441 ,chemistry.chemical_compound ,Adsorption ,food ,Linalool ,Citrus paradisi ,lcsh:Organic chemistry ,Drug Discovery ,hydrodynamic cavitation ,?-terpineol ,Food science ,Physical and Theoretical Chemistry ,IntegroPectin ,pectin ,Residue (complex analysis) ,Limonene ,Volatile Organic Compounds ,Molecular Structure ,applied_chemistry ,Organic Chemistry ,circular economy ,waste citrus peel ,Biosynthetic Pathways ,Terpineol ,chemistry ,Chemistry (miscellaneous) ,Fruit ,Molecular Medicine ,α-terpineol ,Gas chromatography–mass spectrometry ,Citric acid - Abstract
An HS-SPME GC-MS analysis of the volatile compounds adsorbed at the outer surface of lemon and grapefruit pectins obtained via the hydrodynamic cavitation of industrial waste streams of lemon and grapefruit peels in water suggests important new findings en route to understanding the powerful and broad biological activity of these new pectic materials. In agreement with the ultralow degree of esterification of these pectins, the high amount of highly bioactive &alpha, terpineol and terpinen-4-ol points to limonene (and linalool) decomposition catalyzed by residual citric acid in the citrus waste peel residue of the juice industrial production.
- Published
- 2020
23. Seroprevalence of Borrelia burgdorferi in Stray Dogs from Southern Italy
- Author
-
Carmela Sciacca, Giuseppina Chiarenza, Santina Di Bella, Francesco Santangelo, Sonia Sciortino, Alessandra Castiglia, Rosa Alduina, Francesca Grippi, Paola Galluzzo, Maria Arnone, Galluzzo P., Grippi F., Di Bella S., Santangelo F., Sciortino S., Castiglia A., Sciacca C., Arnone M., Alduina R., and Chiarenza G.
- Subjects
0301 basic medicine ,Microbiology (medical) ,030231 tropical medicine ,Biology ,Settore BIO/19 - Microbiologia Generale ,Microbiology ,Serology ,03 medical and health sciences ,0302 clinical medicine ,Lyme disease ,Virology ,medicine ,Seroprevalence ,Borrelia burgdorferi ,lcsh:QH301-705.5 ,Pathogen ,seroprevalence ,Communication ,ospA gene ,Borrelia burgdorferi, seroprevalence ,030108 mycology & parasitology ,medicine.disease ,biology.organism_classification ,bacterial infections and mycoses ,Breed ,lcsh:Biology (General) ,biology.protein ,Ixodes ,Antibody ,stray dogs ,IFA - Abstract
Borrelia burgdorferi is a bacterial pathogen transmitted by Ixodes ticks and is responsible for Lyme disease in both humans and dogs. The aim of this work was to evaluate B. burgdorferi diffusion among stray dogs in Palermo (Sicily, Italy) by serological methods in order to study the risk factors associated with the infection. Serum and blood samples of 316 dogs were collected from a shelter in Palermo, and were analyzed for the presence of antibodies against B. burgdorferi by indirect immunofluorescence assay (IFA), and of the ospA gene by real-time PCR, respectively. Seventeen sera (5.4%) were positive for the antibodies via IFA and one blood (0.3%) for ospA via real time PCR. On the basis of serological results, the evaluation of the potential risk factors (sex, age, breed and coat color) was carried out. The multivariate analysis indicated that male sex is a factor significantly associated with B. burgdorferi seropositivity. This study confirms that male dogs have a higher risk of developing the disease than females, and represents the first investigation on the spread of B. burgdorferi among stray dogs in Sicily.
- Published
- 2020
24. Formulation of Mesoporous Silica Nanoparticles for Controlled Release of Antimicrobials for Stone Preventive Conservation
- Author
-
Alessandro Presentato, Francesco Armetta, Alberto Spinella, Delia Francesca Chillura Martino, Rosa Alduina, Maria Luisa Saladino, Presentato A., Armetta F., Spinella A., Chillura Martino D.F., Alduina R., and Saladino M.L.
- Subjects
Biocide ,condensation in emulsion ,Nanoparticle ,02 engineering and technology ,Bacterial growth ,010402 general chemistry ,Settore BIO/19 - Microbiologia Generale ,01 natural sciences ,Kocuria rhizophila ,biocides ,lcsh:Chemistry ,stone conservation ,Specific surface area ,biodeterioration ,mesoporous silica nanoparticles ,Original Research ,Settore CHIM/02 - Chimica Fisica ,controlled release systems ,biology ,Chemistry ,General Chemistry ,cultural heritage ,Mesoporous silica ,021001 nanoscience & nanotechnology ,biology.organism_classification ,Controlled release ,0104 chemical sciences ,Chemical engineering ,biocides, biodeterioration, condensation in emulsion, controlled release systems, cultural heritage, mesoporous silica nanoparticles, stone conservation ,lcsh:QD1-999 ,Emulsion ,0210 nano-technology - Abstract
The biotic deterioration of artifacts of archaeological and artistic interest mostly relies on the action of microorganisms capable of thriving under the most disparate environmental conditions. Thus, to attenuate biodeterioration phenomena, biocides can be used by the restorers to prevent or slow down the microbial growth. However, several factors such as biocide half-life, its wash-out because of environmental conditions, and its limited time of action make necessary its application repeatedly, leading to negative economic implications. Sound and successful treatments are represented by controlled release systems (CRSs) based on porous materials. Here, we report on the design and development of a CRS system based on mesoporous silica nanoparticles (MSNs), as a carrier, and loaded with a biocide. MSNs, with a diameter of 55 nm and cylindrical pores of ca. 3–8 nm arranged as parallel arrays concerning the NP diameter, and with 422 m2/g of specific surface area were synthesized by the sol-gel method assisted by oil in water emulsion. Biocide loading and release were carried out in water and monitored by UV-Vis Spectroscopy; in addition, microbiological assay was performed using as control the MCM-41 mesoporous silica loaded with the same biocide. The role of specific supramolecular interaction in regulating the release is discussed. Further, we demonstrated that this innovative formulation was useful in inhibiting the in vitro growth of Kocuria rhizophila, an environmental Gram-positive bacterial strain. Besides, the CRS here prepared reduced the bacterial biomass contaminating a real case study (i.e., stone derived from the Santa Margherita cave located in Sicily, Italy), after several months of treatment thus opening for innovative treatments of deteriorated stone artifacts.
- Published
- 2020
25. A combined physical-chemical and microbiological approach to unveil the fabrication, provenance, and state of conservation of the Kinkarakawa-gami art
- Author
-
Antonio Giannusa, Rosa Alduina, Alessandro Presentato, Vittorio Ferrara, Elena Piacenza, Delia Francesca Chillura Martino, Valeria Minore, Francesca Di Salvo, Giuseppe Sancataldo, Piacenza E., Presentato A., Di Salvo F., Alduina R., Ferrara V., Minore V., Giannusa A., Sancataldo G., and Chillura Martino D.F.
- Subjects
0301 basic medicine ,Provenance ,Science ,XRF ,Settore BIO/19 - Microbiologia Generale ,01 natural sciences ,fluorescence microscopy ,Microbiology ,Article ,03 medical and health sciences ,Physical chemical ,Statistical analysis ,Settore CHIM/02 - Chimica Fisica ,Multidisciplinary ,010401 analytical chemistry ,Limiting ,leather-like wallpaper ,cultural heritage ,Materials science ,0104 chemical sciences ,030104 developmental biology ,Geography ,FTIR ,biodeteriogen ,Medicine ,Identification (biology) ,Biochemical engineering - Abstract
Kinkarakawa-gami wallpapers are unique works of art produced in Japan between 1870 and 1905 and exported in European countries, although only few examples are nowadays present in Europe. So far, neither the wallpapers nor the composing materials have been characterised, limiting the effective conservation–restoration of these artefacts accounting also for the potential deteriogen effects of microorganisms populating them. In the present study, four Kinkarakawa-gami wallpapers were analysed combining physical–chemical and microbiological approaches to obtain information regarding the artefacts’ manufacture, composition, dating, and their microbial community. The validity of these methodologies was verified through a fine in blind statistical analysis, which allowed to identify trends and similarities within these important artefacts. The evidence gathered indicated that these wallpapers were generated between 1885 and 1889, during the so-called industrial production period. A wide range of organic (proteinaceous binders, natural waxes, pigments, and vegetable lacquers) and inorganic (tin foil and pigments) substances were used for the artefacts’ manufacture, contributing to their overall complexity, which also reflects on the identification of a heterogeneous microbiota, often found in Eastern environmental matrices. Nevertheless, whether microorganisms inhabiting these wallpapers determined a detrimental or protective effect is not fully elucidated yet, thus constituting an aspect worth to be explored to deepen the knowledge needed for the conservation of Kinkarakawa-gami over time.
- Published
- 2020
26. A Two-Component regulatory system with opposite effects on glycopeptide antibiotic biosynthesis and resistance
- Author
-
Rosa Alduina, Margherita Sosio, Stefano Donadio, Patrizia Cancemi, Clelia Ferraro, Salvatore Costa, Arianna Tocchetti, Alduina R., Tocchetti A., Costa S., Ferraro C., Cancemi P., Sosio M., and Donadio S.
- Subjects
0301 basic medicine ,medicine.drug_class ,030106 microbiology ,lcsh:Medicine ,Glycopeptide antibiotic ,Industrial microbiology ,Article ,03 medical and health sciences ,Bacterial Proteins ,Transcription (biology) ,Genes, Regulator ,Gene cluster ,medicine ,lcsh:Science ,Gene ,Regulator gene ,Regulation of gene expression ,Multidisciplinary ,Antimicrobials ,Chemistry ,lcsh:R ,Gene Expression Regulation, Bacterial ,Glycopeptide ,Anti-Bacterial Agents ,Biosynthetic Pathways ,Cell biology ,Actinobacteria ,Response regulator ,030104 developmental biology ,Multigene Family ,Two component regulatory system, glycopeptide A40926, actinomycete Nonomuraea gerenzanensis ,lcsh:Q ,Teicoplanin ,Microbial genetics - Abstract
The glycopeptide A40926, produced by the actinomycete Nonomuraea gerenzanensis, is the precursor of dalbavancin, a second-generation glycopeptide antibiotic approved for clinical use in the USA and Europe in 2014 and 2015, respectively. The final product of the biosynthetic pathway is an O-acetylated form of A40926 (acA40926). Glycopeptide biosynthesis in N. gerenzanensis is dependent upon the dbv gene cluster that encodes, in addition to the two essential positive regulators Dbv3 and Dbv4, the putative members of a two-component signal transduction system, specifically the response regulator Dbv6 and the sensor kinase Dbv22. The aim of this work was to assign a role to these two genes. Our results demonstrate that deletion of dbv22 leads to an increased antibiotic production with a concomitant reduction in glycopeptide resistance. Deletion of dbv6 results in a similar phenotype, although the effects are not as strong as in the Δdbv22 mutant. Consistently, quantitative RT-PCR analysis showed that Dbv6 and Dbv22 negatively regulate the regulatory genes (dbv3 and dbv4), as well as some dbv biosynthetic genes (dbv23 and dbv24), whereas Dbv6 and Dbv22 positively regulate transcription of the single, cluster-associated resistance gene. Finally, we demonstrate that exogenously added acA40926 and its precursor A40926 can modulate transcription of dbv genes but with an opposite extent: A40926 strongly stimulates transcription of the Dbv6/Dbv22 target genes while acA40926 has a neutral or negative effect on transcription of those genes. We propose a model in which glycopeptide biosynthesis in N. gerenzanensis is modulated through a positive feedback by the biosynthetic precursor A40926 and a negative feedback by the final product acA40926. In addition to previously reported control systems, this sophisticated control loop might help the producing strain cope with the toxicity of its own product. This work, besides leading to improved glycopeptide producing strains, enlarges our knowledge on the regulation of glycopeptide biosynthesis in actinomycetes, setting N. gerenzanensis and its two-component system Dbv6-Dbv22 apart from other glycopeptide producers.
- Published
- 2020
27. Antibiotic Resistance of Gram-Negative Bacteria from Wild Captured Loggerhead Sea Turtles
- Author
-
Monica Francesca Blasi, Rosa Alduina, Luciana Migliore, Alice Rotini, Maria Cristina Thaller, Daniela Mattei, Blasi M.F., Migliore L., Mattei D., Rotini A., Thaller M.C., and Alduina R.
- Subjects
0301 basic medicine ,Microbiology (medical) ,Gram-negative bacteria ,antibiotic resistance ,Settore BIO/07 ,030106 microbiology ,Zoology ,Biochemistry ,Microbiology ,Article ,bacterial ecology ,03 medical and health sciences ,Marine bacteriophage ,Mediterranean sea ,Antibiotic resistance ,Vibrionaceae ,Caretta caretta ,Mediterranean Sea ,feeding ,marine bacteria ,marine habitats ,marine microbial ecology ,Ampicillin ,medicine ,Pharmacology (medical) ,General Pharmacology, Toxicology and Pharmaceutics ,biology ,Sulfamethoxazole ,lcsh:RM1-950 ,biology.organism_classification ,030104 developmental biology ,Infectious Diseases ,lcsh:Therapeutics. Pharmacology ,marine habitat ,Bacteria ,medicine.drug - Abstract
Sea turtles have been proposed as health indicators of marine habitats and carriers of antibiotic-resistant bacterial strains, for their longevity and migratory lifestyle. Up to now, a few studies evaluated the antibacterial resistant flora of Mediterranean loggerhead sea turtles (Caretta caretta) and most of them were carried out on stranded or recovered animals. In this study, the isolation and the antibiotic resistance profile of 90 Gram negative bacteria from cloacal swabs of 33 Mediterranean wild captured loggerhead sea turtles are described. Among sea turtles found in their foraging sites, 23 were in good health and 10 needed recovery for different health problems (hereafter named weak). Isolated cloacal bacteria belonged mainly to Enterobacteriaceae (59%), Shewanellaceae (31%) and Vibrionaceae families (5%). Although slight differences in the bacterial composition, healthy and weak sea turtles shared antibiotic-resistant strains. In total, 74 strains were endowed with one or multi resistance (up to five different drugs) phenotypes, mainly towards ampicillin (~70%) or sulfamethoxazole/trimethoprim (more than 30%). Hence, our results confirmed the presence of antibiotic-resistant strains also in healthy marine animals and the role of the loggerhead sea turtles in spreading antibiotic-resistant bacteria.
- Published
- 2020
28. Is Caretta Caretta a Carrier of Antibiotic Resistance in the Mediterranean Sea?
- Author
-
Dario Savoca, Rosa Alduina, Giulia Caracappa, Serena Filippello, Marco Arculeo, Antonino Gentile, Giulia Visconti, Domenico Vicari, Arianna Sucato, Delia Gambino, Alessandro Presentato, Alduina R., Gambino D., Presentato A., Gentile A., Sucato A., Savoca D., Filippello S., Visconti G., Caracappa G., Vicari D., and Arculeo M.
- Subjects
0301 basic medicine ,Microbiology (medical) ,antibiotic resistance ,030106 microbiology ,Zoology ,heavy metal resistance ,Integron ,Antibiotic resistance Caretta caretta Mediterranean Sea ,Biochemistry ,Microbiology ,Loggerhead sea turtle ,Article ,antimicrobials ,03 medical and health sciences ,Antibiotic resistance ,Mediterranean sea ,caretta caretta ,medicine ,Mediterranean Sea ,Pharmacology (medical) ,Marine ecosystem ,General Pharmacology, Toxicology and Pharmaceutics ,biology ,mobile element ,fungi ,lcsh:RM1-950 ,biology.organism_classification ,loggerhead sea turtle ,030104 developmental biology ,Infectious Diseases ,lcsh:Therapeutics. Pharmacology ,Aeromonas ,biology.protein ,Colistin ,Seawater ,medicine.drug - Abstract
Sea turtles can be considered a sentinel species for monitoring the health of marine ecosystems, acting, at the same time, as a carrier of microorganisms. Indeed, sea turtles can acquire the microbiota from their reproductive sites and feeding, contributing to the diffusion of antibiotic-resistant strains to uncontaminated environments. This study aims to unveil the presence of antibiotic-resistant bacteria in (i) loggerhead sea turtles stranded along the coast of Sicily (Mediterranean Sea), (ii) unhatched and/or hatched eggs, (iii) sand from the turtles&rsquo, nest and (iv) seawater. Forty-four bacterial strains were isolated and identified by conventional biochemical tests and 16S rDNA sequencing. The Gram-negative Aeromonas and Vibrio species were mainly found in sea turtles and seawater samples, respectively. Conversely, the Gram-positive Bacillus, Streptococcus, and Staphylococcus strains were mostly isolated from eggs and sand. The antimicrobial resistance profile of the isolates revealed that these strains were resistant to cefazolin (95.5%), streptomycin (43.2%), colistin and amoxicillin/clavulanic acid (34.1%). Moreover, metagenome analysis unveiled the presence of both antibiotic and heavy metal resistance genes, as well as the mobile element class 1 integron at an alarming percentage rate. Our results suggest that Caretta caretta could be considered a carrier of antibiotic-resistant genes.
- Published
- 2020
29. Lipid Nanocarriers-Loaded Nanocomposite as a Suitable Platform to Release Antibacterial and Antioxidant Agents for Immediate Dental Implant Placement Restorative Treatment
- Author
-
Giuseppe Angellotti, Alessandro Presentato, Denise Murgia, Giulia Di Prima, Fabio D’Agostino, Amalia Giulia Scarpaci, Maria Cristina D’Oca, Rosa Alduina, Giuseppina Campisi, Viviana De Caro, Angellotti G., Presentato A., Murgia D., Di Prima G., D'Agostino F., Scarpaci A.G., D'Oca M.C., Alduina R., Campisi G., and De Caro V.
- Subjects
Chitosan ,Nanocomposite ,Nanostructured lipid carriers ,Pharmaceutical Science ,Ex vivo permeation ,Settore BIO/19 - Microbiologia Generale ,Article ,Antibiofilm ,RS1-441 ,Membrane accumulation ,Pharmacy and materia medica ,Ciprofloxacin ,Settore CHIM/09 - Farmaceutico Tecnologico Applicativo ,nanocomposite ,quercetin ,ciprofloxacin ,nanostructured lipid carriers ,chitosan ,antimicrobial ,antioxidant ,antibiofilm ,ex vivo permeation ,membrane accumulation ,Antimicrobial ,Quercetin ,Antioxidant - Abstract
Immediate implant placement is a single-stage restorative approach for missing teeth widely used to overcome the ridge remodeling process occurring after dental extractions. The success of this procedure relies on opportune osseointegration in the surrounding tissues. To support this process, a multifunctional nanocomposite, to be applied in the fresh post-extraction socket, was here designed, prepared, and characterized. This formulation consists of quercetin (QRC)-loaded nanostructured lipid carriers (NLCs) entrapped in a chitosan-based solid matrix containing ciprofloxacin (CPX). QRC-NLCs were prepared by homogenization followed by high-frequency sonication, and thereafter this dispersion was trapped in a chitosan-based CPX-loaded gel, obtaining the nanocomposite powder (BioQ-CPX) by lyophilization. BioQ-CPX displayed desirable properties such as high porosity (94.1 ± 0.5%), drug amounts (2.1% QRC and 3.5% CPX). and low swelling index (100%). Moreover, the mechanism of drug release from BioQ-CPX and their ability to be accumulated in the target tissue were in vitro and ex vivo elucidated, also by applying mathematical models. When trapped into the nanocomposite, QRC stressed under UV light exposure (50 W) was shown to maintain its antioxidant power, and CPX and QRC under natural light were stable over nine months. Finally, both the measured antioxidant power and the antimicrobial and antibiofilm properties on Staphylococcus aureus demonstrated that BioQ-CPX could be a promising platform to support the single-stage dental restorative treatment.
- Published
- 2021
30. Biogenic Selenium Nanoparticles: A Fine Characterization to Unveil Their Thermodynamic Stability
- Author
-
Delia Francesca Chillura Martino, Francesco Ferrante, Alessandro Presentato, Giuseppe Cavallaro, Rosa Alduina, Elena Piacenza, Piacenza E., Presentato A., Ferrante F., Cavallaro G., Alduina R., and Chillura Martino D.F.
- Subjects
Biocompatibility ,General Chemical Engineering ,Nanoparticle ,02 engineering and technology ,DFT calculations ,Article ,Micrococcus ,Nanomaterials ,03 medical and health sciences ,Adsorption ,biogenic selenium nanoparticles ,Molecule ,General Materials Science ,Fourier transform infrared spectroscopy ,QD1-999 ,030304 developmental biology ,multivariate statistical analysis ,chemistry.chemical_classification ,0303 health sciences ,Biomolecule ,021001 nanoscience & nanotechnology ,Chemistry ,FTIR spectroscopy ,chemistry ,Chemical engineering ,thermodynamic stability ,Chemical stability ,selenium nanorods ,0210 nano-technology - Abstract
Among the plethora of available metal(loid) nanomaterials (NMs), those containing selenium are interesting from an applicative perspective, due to their high biocompatibility. Microorganisms capable of coping with toxic Se-oxyanions generate mostly Se nanoparticles (SeNPs), representing an ideal and green alternative over the chemogenic synthesis to obtain thermodynamically stable NMs. However, their structural characterization, in terms of biomolecules and interactions stabilizing the biogenic colloidal solution, is still a black hole that impairs the exploitation of biogenic SeNP full potential. Here, spherical and thermodynamically stable SeNPs were produced by a metal(loid) tolerant Micrococcus sp. Structural characterization obtained by Scanning Electron Microscopy (SEM) revealed that these SeNPs were surrounded by an organic material that contributed the most to their electrosteric stabilization, as indicated by Zeta (ζ) potential measurements. Proteins were strongly adsorbed on the SeNP surface, while lipids, polysaccharides, and nucleic acids more loosely interacted with SeNMs as highlighted by Fourier Transform Infrared Spectroscopy (FTIR) and overall supported by multivariate statistical analysis. Nevertheless, all these contributors were fundamental to maintain SeNPs stable, as, upon washing, the NM-containing extract showed the arising of aggregated SeNPs alongside Se nanorods (SeNRs). Besides, Density Functional Theory (DFT) calculation unveiled how thiol-containing molecules appeared to play a role in SeO32− bioreduction, stress oxidative response, and SeNP stabilization.
- Published
- 2021
31. Occurrence and Antimicrobial Resistance of Arcobacter spp. Recovered from Aquatic Environments
- Author
-
Sonia Sciortino, Vincenzina Alio, Rosa Alduina, C. Cardamone, Pietro Arculeo, Antonella Costa, Marco Arculeo, Luisa Nicastro, Sciortino S., Arculeo P., Alio V., Cardamone C., Nicastro L., Arculeo M., Alduina R., and Costa A.
- Subjects
0301 basic medicine ,Microbiology (medical) ,Water samples ,Veterinary medicine ,Antibiotic susceptibility ,Arcobacter cryaerophilus ,TetW ,Tetracycline ,medicine.drug_class ,030106 microbiology ,Antibiotics ,TetO ,Biochemistry ,Microbiology ,Article ,03 medical and health sciences ,Antibiotic resistance ,medicine ,Pharmacology (medical) ,General Pharmacology, Toxicology and Pharmaceutics ,biology ,Aquatic ecosystem ,lcsh:RM1-950 ,Arcobacter butzleri ,Multiplex PCR ,biology.organism_classification ,Isolation (microbiology) ,lcsh:Therapeutics. Pharmacology ,030104 developmental biology ,Infectious Diseases ,Arcobacter ,tetO ,tetW ,medicine.drug - Abstract
Arcobacter spp. are emerging waterborne and foodborne zoonotic pathogens responsible for gastroenteritis in humans. In this work, we evaluated the occurrence and the antimicrobial resistance profile of Arcobacter isolates recovered from different aquatic sources. Besides, we searched for Arcobacter spp. in seaweeds and the corresponding seawater samples. Bacteriological and molecular methods applied to 100 samples led to the isolation of 28 Arcobacter isolates from 27 samples. The highest prevalence was detected in rivers followed by artificial ponds, streams, well waters, and spring waters. Seaweeds contained a higher percentage of Arcobacter than the corresponding seawater samples. The isolates were identified as Arcobacter butzleri (96.4%) and Arcobacter cryaerophilus (3.6%). All the isolates showed a multi-drug resistance profile, being resistant to at least three different classes of antibiotics. Molecular analysis of genetic determinants responsible for tetracycline resistance in nine randomly chosen isolates revealed the presence of tetO and/or tetW. This work confirms the occurrence and the continuous emergence of antibiotic-resistant Arcobacter strains in environmental samples, also, the presence of quinolone-resistant Arcobacter spp. in aquatic sources used for water supply and irrigation represents a potential risk for human health.
- Published
- 2021
32. A Comparative Analysis of Aquatic and Polyethylene-Associated Antibiotic-Resistant Microbiota in the Mediterranean Sea
- Author
-
Alessandro Presentato, Arianna Sucato, Rosa Alduina, Luca Vecchioni, Marco Arculeo, Dario Savoca, Sucato A., Vecchioni L., Savoca D., Presentato A., Arculeo M., and Alduina R.
- Subjects
0301 basic medicine ,Aquatic environments ,030106 microbiology ,Plastisphere ,Biology ,Integron ,Article ,Resistome ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,Antibiotic resistance ,Mediterranean sea ,Antibiotic resistance genes ,Food science ,Microbiome ,lcsh:QH301-705.5 ,General Immunology and Microbiology ,Aquatic ecosystem ,030104 developmental biology ,lcsh:Biology (General) ,Antibiotic resistant bacteria ,Polyethylene ,biology.protein ,Seawater ,General Agricultural and Biological Sciences - Abstract
Simple Summary In recent years, a growing interest has been devoted to the bacterial characterization of marine plastic debris. So far, a few publications have explored the composition of microbial communities on polyethylene (PE) waste items and the occurrence of antibiotic-resistant bacteria (ARB). The occurrence of ARB in natural matrices can contribute to the spread of antibiotic resistance genes (ARGs) among environmental bacteria. In this study, we compared the microbial composition and the presence of ARGs in water and PE fragments collected from a stream and the seawater in a coastal area of Northwestern Sicily. Our findings showed more ARGs on PE fragments than the corresponding water samples, confirming that PE can act as a carrier of antibiotic-resistance genes causing high damage to the marine environment and living organisms. Abstract In this study, we evaluated the microbiome and the resistome profile of water and fragments of polyethylene (PE) waste collected at the same time from a stream and the seawater in a coastal area of Northwestern Sicily. Although a core microbiome was determined by sequencing of the V3–V4 region of the bacterial 16S rDNA gene, quantitative differences were found among the microbial communities on PE waste and the corresponding water samples. Our findings indicated that PE waste contains a more abundant and increased core microbiome diversity than the corresponding water samples. Moreover, PCR analysis of specific antibiotic resistance genes (ARGs) showed that PE waste harbors more ARGs than the water samples. Thus, PE waste could act as a carrier of antibiotic-resistant microbiota, representing an increased danger for the marine environment and living organisms, as well.
- Published
- 2021
33. Mononuclear Perfluoroalkyl-Heterocyclic Complexes of Pd(II): Synthesis, Structural Characterization and Antimicrobial Activity
- Author
-
Vita Di Stefano, Patrizia Cancemi, Rosa Alduina, Maria Assunta Girasolo, Silvestre Buscemi, Ivana Pibiri, Simona Rubino, Santino Orecchio, Rubino S., Alduina R., Cancemi P., Girasolo M.A., Di Stefano V., Orecchio S., Buscemi S., and Pibiri I.
- Subjects
Denticity ,perfluoroalkyl heterocyclic ligands ,Spectrophotometry, Infrared ,Stereochemistry ,Proton Magnetic Resonance Spectroscopy ,Pharmaceutical Science ,Microbial Sensitivity Tests ,Settore BIO/19 - Microbiologia Generale ,Ring (chemistry) ,Analytical Chemistry ,lcsh:QD241-441 ,chemistry.chemical_compound ,lcsh:Organic chemistry ,Anti-Infective Agents ,Heterocyclic Compounds ,Drug Discovery ,Pyridine ,mononuclear palladium complexes ,Settore BIO/06 - Anatomia Comparata E Citologia ,Physical and Theoretical Chemistry ,triazoles ,Fluorocarbons ,antimicrobial activity ,Bacteria ,Chemistry ,Ligand ,Communication ,narcosis ,Organic Chemistry ,Settore CHIM/06 - Chimica Organica ,DNA ,Antimicrobial ,Settore CHIM/03 - Chimica Generale E Inorganica ,Chemistry (miscellaneous) ,Molecular Medicine ,Palladium ,Plasmids - Abstract
Two mononuclear Pd(II) complexes [PdCl2(pfptp)] (1) and [PdCl2(pfhtp)] (2), with ligands 2-(3-perfluoropropyl-1-methyl-1,2,4-triazole-5yl)-pyridine (pfptp) and 2-(3-perfluoroheptyl-1-methyl-1,2,4-triazole-5yl)-pyridine (pfhtp), were synthesized and structurally characterized. The two complexes showed a bidentate coordination of the ligand occurring through N atom of pyridine ring and N4 atom of 1,2,4-triazole. Both complexes showed antimicrobial activity when tested against both Gram-negative and Gram-positive bacterial strains.
- Published
- 2020
34. Antibiotic Resistance Profiling, Analysis of Virulence Aspects and Molecular Genotyping of Staphylococcus aureus Isolated in Sicily, Italy
- Author
-
Salvatore Gaglio, Chiara Piraino, Vincenzo Di Marco Lo Presti, Giuseppe Cascone, Maria Vitale, Rosa Alduina, Paola Galluzzo, Vitale, M., Gaglio, S., Galluzzo, P., Cascone, G., Piraino, C., Di Marco Lo Presti, V., and Alduina, R.
- Subjects
0301 basic medicine ,Staphylococcus aureus ,Livestock ,antibiotic resistance ,Genotype ,MLST ,MRSA ,PFGE ,toxin genes ,Tetracycline ,030106 microbiology ,Virulence ,Biology ,medicine.disease_cause ,Settore BIO/19 - Microbiologia Generale ,Applied Microbiology and Biotechnology ,Microbiology ,Foodborne Diseases ,Enterotoxins ,03 medical and health sciences ,Antibiotic resistance ,Drug Resistance, Bacterial ,Pulsed-field gel electrophoresis ,medicine ,Animals ,Humans ,Sicily ,Retrospective Studies ,Outbreak ,Original Articles ,Staphylococcal Infections ,Anti-Bacterial Agents ,Bacterial Typing Techniques ,Electrophoresis, Gel, Pulsed-Field ,Penicillin ,030104 developmental biology ,Staphylococcus aureu ,Food Microbiology ,Multilocus sequence typing ,Animal Science and Zoology ,Multilocus Sequence Typing ,Food Science ,medicine.drug - Abstract
Staphylococcus aureus is the major cause of foodborne diseases worldwide. In this retrospective study, 84 S. aureus strains were characterized. The collection comprises 78 strains isolated during 1998 and 2014 from dairy products and tissue samples from livestock bred for dairy production in Sicily. One isolate was obtained from a pet (dog), one from an exotic animal (a circus elephant), and four human isolates were obtained during a severe food poisoning outbreak that occurred in Sicily in 2015. All the strains were characterized by pulsed-field gel electrophoresis (PFGE), for antibiotic resistance and presence of toxin genes. PFGE results showed 10 different pulsotypes, with three relatively frequent and three unique. The antibiotic resistance profiling showed that penicillin G (35.7%) and tetracycline (20.2%) resistance is largely spread. Most isolates contained at least one toxin gene making them a potential threat for public health. Enterotoxin sec gene was observed in 28.6% and seg in 23.8% of the strains, respectively; the human isolates were the only ones to concurrently harbor both seg and sei genes. In addition, 24 isolates were randomly selected and analyzed by multilocus sequence typing. Interestingly, the analysis showed the presence of 12 sequence types (STs), of which 6 were novel. One of them, ST700, was detected in 29% of the isolates and was found to be spread throughout Sicily. ST700 has been present in the island for almost 16 years (1998–2014) and it shows no host preference since it was isolated from different ruminant species. Four human isolates shared both the pulsotype (PT10) and the sequence type (ST9), as well as the virulence genes (seg-sei); this observation suggests that the isolates originated from a single clone, although they were obtained from two different individuals.
- Published
- 2018
35. Binding abilities of polyaminocyclodextrins: polarimetric investigations and biological assays
- Author
-
Daniele La Corte, Paolo Lo Meo, Marco Russo, Rosa Alduina, Serena Riela, Annalisa Pisciotta, Russo, M., La Corte, D., Pisciotta, A., Riela, S., Alduina, R., and Lo Meo, P.
- Subjects
nitroaniline ,aminocyclodextrins ,binding properties ,nitroanilines ,pDNA ,polarimetry ,supramolecular chemistry ,Supramolecular chemistry ,010402 general chemistry ,01 natural sciences ,Full Research Paper ,aminocyclodextrin ,lcsh:QD241-441 ,lcsh:Organic chemistry ,Bioassay ,binding propertie ,lcsh:Science ,010405 organic chemistry ,Chemistry ,Binding properties ,Organic Chemistry ,Cationic polymerization ,0104 chemical sciences ,Polynucleotide ,Biophysics ,lcsh:Q ,pUC19 ,Direct reaction ,Macromolecule - Abstract
Three polyaminocyclodextrin materials, obtained by direct reaction between heptakis(6-deoxy-6-iodo)-β-cyclodextrin and the proper linear polyamines, were investigated for their binding properties, in order to assess their potential applications in biological systems, such as vectors for simultaneous drug and gene cellular uptake or alternatively for the protection of macromolecules. In particular, we exploited polarimetry to test their interaction with some model p-nitroaniline derivatives, chosen as probe guests. The data obtained indicate that binding inside the host cavity is mainly affected by interplay between Coulomb interactions and conformational restraints. Moreover, simultaneous interaction of the cationic polyamine pendant bush at the primary rim was positively assessed. Insights on quantitative aspects of the interaction between our materials and polyanions were investigated by studying the binding with sodium alginate. Finally, the complexation abilities of the same materials towards polynucleotides were assessed by studying their interaction with the model plasmid pUC19. Our results positively highlight the ability of our materials to exploit both the cavity and the polycationic branches, thus functioning as bimodal ligands.
- Published
- 2017
36. Synthesis and antibacterial activity of iron-hexacyanocobaltate nanoparticles
- Author
-
Eugenio Caponetti, Maria Luisa Saladino, Michela Ciabocco, Patrizia Cancemi, Rosa Alduina, Mario Berrettoni, Ciabocco, Michela, Cancemi, Patrizia, Saladino, Maria Luisa, Caponetti, Eugenio, Alduina, Rosa, Berrettoni, Mario, and Ciabocco M, Cancemi P, Saladino ML, Caponetti E, Alduina R, Berrettoni M
- Subjects
Metal-hexacyanoferrate ,Staphylococcus aureus ,Iron ,Colony Count, Microbial ,Infrared spectroscopy ,Nanoparticle ,Metal Nanoparticles ,02 engineering and technology ,Microbial Sensitivity Tests ,Bacterial growth ,010402 general chemistry ,medicine.disease_cause ,01 natural sciences ,Biochemistry ,Fluorescence spectroscopy ,Inorganic Chemistry ,Microscopy, Electron, Transmission ,medicine ,Fluorescence microscope ,Escherichia coli ,Cyanides ,Chemistry ,Iron-hexacyanocobaltate ,Cobalt ,021001 nanoscience & nanotechnology ,0104 chemical sciences ,Anti-Bacterial Agents ,Spectrometry, Fluorescence ,Staphylococcus aureu ,Microscopy, Electron, Scanning ,Antibacterial activity ,0210 nano-technology ,Reactive Oxygen Species ,Nuclear chemistry ,Macromolecule - Abstract
This paper deals with the synthesis and characterization of iron-hexacyanocobaltate (FeHCC) and its antibacterial properties. The nanoparticles were prepared by a facile co-precipitation technique. Crystal structure, particle morphology, and elemental composition were determined using X-ray Powder Diffraction, X-ray fluorescence spectroscopy, Transmission Electron Microscopy (TEM), and Infrared Spectroscopy (IR). The antibacterial activity of the FeHCC nanoparticles was tested against Escherichia coli and Staphylococcus aureus as models for Gram-negative and Gram-positive bacteria, respectively, by bacterial counting method and microscopic visualization (TEM, FEG-SEM, and fluorescence microscopy). The results showed that the FeHCC nanoparticles bind to the bacterial cells, inhibit bacterial growth in a dose- and time-dependent manner, inducing a loss of the membrane potential, the production of reactive oxygen species and the release of macromolecules (nucleic acids and proteins) in the extracellular environment. To the best of our knowledge, this is the first study reporting the antimicrobial effects of metal-hexacyanometallates suggesting practical uses of these materials in different areas, such as self-cleaning surfaces or food packaging.
- Published
- 2017
37. New ΦBT1 site-specific integrative vectors with neutral phenotype in Streptomyces
- Author
-
Annalisa Pisciotta, María Teresa López-García, Angel Manteca, Nathaly Gonzalez-Quiñonez, Paula Yagüe, Beatriz Rioseras, Rosa Alduina, Gonzalez-Quiñonez, N., López-García, M., Yagüe, P., Rioseras, B., Pisciotta, A., Alduina, R., and Manteca, Á.
- Subjects
0301 basic medicine ,Genetics, Microbial ,030106 microbiology ,Genetic Vectors ,Settore BIO/19 - Microbiologia Generale ,Genome ,Streptomyces ,Applied Microbiology and Biotechnology ,03 medical and health sciences ,Plasmid ,Amp resistance ,Spore germination ,Escherichia coli ,Neutral phenotype ,Bacteriophages ,Vector (molecular biology) ,Gene ,Molecular Biology ,Genetics ,Recombination, Genetic ,biology ,fungi ,General Medicine ,biology.organism_classification ,Phenotype ,ΦBT1 integrative vector ,030104 developmental biology ,Streptomyce ,Heterologous expression ,Biotechnology ,Plasmids - Abstract
Integrative plasmids are one of the best options to introduce genes in low copy and in a stable form into bacteria. The ΦC31-derived plasmids constitute the most common integrative vectors used in Streptomyces. They integrate at different positions (attB and pseudo-attB sites) generating different mutations. The less common ΦBT1-derived vectors integrate at the unique attB site localized in the SCO4848 gene (S. coelicolor genome) or their orthologues in other streptomycetes. This work demonstrates that disruption of SCO4848 generates a delay in spore germination. SCO4848 is co-transcribed with SCO4849, and the spore germination phenotype is complemented by SCO4849. Plasmids pNG1-4 were created by modifying the ΦBT1 integrative vector pMS82 by introducing a copy of SCO4849 under the control of the promoter region of SCO4848. pNG2 and pNG4 also included a copy of the P ermE * in order to facilitate gene overexpression. pNG3 and pNG4 harboured a copy of the bla gene (ampicillin resistance) to facilitate selection in E. coli. pNG1-4 are the only integrative vectors designed to produce a neutral phenotype when they are integrated into the Streptomyces genome. The experimental approach developed in this work can be applied to create phenotypically neutral integrative plasmids in other bacteria.
- Published
- 2015
38. Two master switch regulators trigger A40926 biosynthesis in Nonomuraea sp. strain ATCC 39727
- Author
-
Margherita Sosio, Rosa Alduina, Letizia Lo Grasso, Mervyn J. Bibb, Sonia I. Maffioli, Anna Maria Puglia, Lo Grasso, L, Maffioli, S, Sosio, M, Bibb, M, Puglia, A.M, and Alduina, R
- Subjects
Transcription, Genetic ,Operon ,medicine.drug_class ,Biology ,Glycopeptide antibiotic ,Settore BIO/19 - Microbiologia Generale ,Microbiology ,chemistry.chemical_compound ,Bacterial Proteins ,Biosynthesis ,Transcription (biology) ,Actinomycetales ,Gene cluster ,medicine ,A40926 Biosynthesi ,Molecular Biology ,Gene ,Regulation of gene expression ,Molecular Structure ,Reverse Transcriptase Polymerase Chain Reaction ,Gene Expression Regulation, Bacterial ,Articles ,Anti-Bacterial Agents ,Biochemistry ,chemistry ,Mannosylation ,Mutation ,Nonomuraea sp. Strain ATCC 39727 ,gene expression ,Teicoplanin - Abstract
The actinomycete Nonomuraea sp. strain ATCC 39727 produces the glycopeptide A40926, the precursor of dalbavancin. Biosynthesis of A40926 is encoded by the dbv gene cluster, which contains 37 protein-coding sequences that participate in antibiotic biosynthesis, regulation, immunity, and export. In addition to the positive regulatory protein Dbv4, the A40926-biosynthetic gene cluster encodes two additional putative regulators, Dbv3 and Dbv6. Independent mutations in these genes, combined with bioassays and liquid chromatography-mass spectrometry (LC-MS) analyses, demonstrated that Dbv3 and Dbv4 are both required for antibiotic production, while inactivation of dbv6 had no effect. In addition, overexpression of dbv3 led to higher levels of A40926 production. Transcriptional and quantitative reverse transcription (RT)-PCR analyses showed that Dbv4 is essential for the transcription of two operons, dbv14-dbv8 and dbv30-dbv35 , while Dbv3 positively controls the expression of four monocistronic transcription units ( dbv4 , dbv29 , dbv36 , and dbv37 ) and of six operons ( dbv2-dbv1 , dbv14-dbv8 , dbv17-dbv15 , dbv21-dbv20 , dbv24-dbv28 , and dbv30-dbv35 ). We propose a complex and coordinated model of regulation in which Dbv3 directly or indirectly activates transcription of dbv4 and controls biosynthesis of 4-hydroxyphenylglycine and the heptapeptide backbone, A40926 export, and some tailoring reactions (mannosylation and hexose oxidation), while Dbv4 directly regulates biosynthesis of 3,5-dihydroxyphenylglycine and other tailoring reactions, including the four cross-links, halogenation, glycosylation, and acylation. IMPORTANCE This report expands knowledge of the regulatory mechanisms used to control the biosynthesis of the glycopeptide antibiotic A40926 in the actinomycete Nonomuraea sp. strain ATCC 39727. A40926 is the precursor of dalbavancin, approved for treatment of skin infections by Gram-positive bacteria. Therefore, understanding the regulation of its biosynthesis is also of industrial importance. So far, the regulatory mechanisms used to control two other similar glycopeptides (balhimycin and teicoplanin) have been elucidated, and beyond a common step, different clusters seem to have devised different strategies to control glycopeptide production. Thus, our work provides one more example of the pitfalls of deducing regulatory roles from bioinformatic analyses only, even when analyzing gene clusters directing the synthesis of structurally related compounds.
- Published
- 2015
39. Elucidating the molecular physiology of lantibiotic NAI-107 production in Microbispora ATCC-PTA-5024
- Author
-
Tilmann Weber, A. Russo, Giovanni Spinelli, Anna Giardina, E. Palazzotto, Margherita Sosio, Rosa Alduina, Giuseppe Gallo, Simona Arena, Anna Maria Puglia, Fabio Sangiorgi, Andrea Scaloni, Giovanni Renzone, T. Faddetta, Paolo Monciardini, Gallo, G., Renzone, G., Palazzotto, E., Monciardini, P., Arena, S., Faddetta, T., Giardina, A., Alduina, R., Weber, T., Sangiorgi, F., Russo, A., Spinelli, G., Sosio, M., Scaloni, A., and Puglia, A.
- Subjects
0301 basic medicine ,Proteomics ,food.ingredient ,Metabolic network ,ATP-binding cassette transporter ,Actinomycetes Antibiotic production Differential proteomics 2D-DIGE and mass spectrometry Metabolic pathways Regulatory network Molecular and cellular functions ,Biology ,Bioinformatics ,Gram-Positive Bacteria ,03 medical and health sciences ,food ,Bacteriocins ,Actinomycetes ,Genetics ,2D-DIGE and mass spectrometry ,Differential proteomics ,2. Zero hunger ,Gel electrophoresis ,Lipid metabolism ,Regulatory network ,biology.organism_classification ,Drug Resistance, Multiple ,Anti-Bacterial Agents ,Actinobacteria ,Metabolic pathway ,030104 developmental biology ,Biochemistry ,Microbispora ,Metabolic pathways ,ATP-Binding Cassette Transporters ,Antibiotic production ,Peptides ,Bacteria ,Molecular and cellular functions ,Biotechnology ,Research Article - Abstract
Background The filamentous actinomycete Microbispora ATCC-PTA-5024 produces the lantibiotic NAI-107, which is an antibiotic peptide effective against multidrug-resistant Gram-positive bacteria. In actinomycetes, antibiotic production is often associated with a physiological differentiation program controlled by a complex regulatory and metabolic network that may be elucidated by the integration of genomic, proteomic and bioinformatic tools. Accordingly, an extensive evaluation of the proteomic changes associated with NAI-107 production was performed on Microbispora ATCC-PTA-5024 by combining two-dimensional difference in gel electrophoresis, mass spectrometry and gene ontology approaches. Results Microbispora ATCC-PTA-5024 cultivations in a complex medium were characterized by stages of biomass accumulation (A) followed by biomass yield decline (D). NAI-107 production started at 90 h (A stage), reached a maximum at 140 h (D stage) and decreased thereafter. To reveal patterns of differentially represented proteins associated with NAI-107 production onset and maintenance, differential proteomic analyses were carried-out on biomass samples collected: i) before (66 h) and during (90 h) NAI-107 production at A stage; ii) during three time-points (117, 140, and 162 h) at D stage characterized by different profiles of NAI-107 yield accumulation (117 and 140 h) and decrement (162 h). Regulatory, metabolic and unknown-function proteins, were identified and functionally clustered, revealing that nutritional signals, regulatory cascades and primary metabolism shift-down trigger the accumulation of protein components involved in nitrogen and phosphate metabolism, cell wall biosynthesis/maturation, lipid metabolism, osmotic stress response, multi-drug resistance, and NAI-107 transport. The stimulating role on physiological differentiation of a TetR-like regulator, originally identified in this study, was confirmed by the construction of an over-expressing strain. Finally, the possible role of cellular response to membrane stability alterations and of multi-drug resistance ABC transporters as additional self-resistance mechanisms toward the lantibiotic was confirmed by proteomic and confocal microscopy experiments on a Microbispora ATCC-PTA-5024 lantibiotic-null producer strain which was exposed to an externally-added amount of NAI-107 during growth. Conclusion This study provides a net contribution to the elucidation of the regulatory, metabolic and molecular patterns controlling physiological differentiation in Microbispora ATCC-PTA-5024, supporting the relevance of proteomics in revealing protein players of antibiotic biosynthesis in actinomycetes. Electronic supplementary material The online version of this article (doi:10.1186/s12864-016-2369-z) contains supplementary material, which is available to authorized users.
- Published
- 2015
40. Epigenetic control of Streptomyces coelicolor differentiation
- Author
-
PISCIOTTA, Annalisa, ALDUINA, Rosa, Galardini, M, Manteca, A, Pisciotta, A, Galardini, M, Manteca, A, and Alduina, R
- Subjects
DNA methylation, Streptomyces coelicolor, epigenetic - Abstract
DNA cytosine methylation is one of the most important epigenetic modifications in eukaryotes regulating chromatin organization, genome maintenance and gene expression. The role of DNA cytosine methylation in prokaryotes has not been deeply investigated. In Escherichia coli cytosine methylation regulates gene expression during the stationary phase and cytosine hypermethylation leads to chromosomal DNA cleavage and cell death. Streptomyces coelicolor is a mycelial soil microorganism, which exhibits a complex life cycle that includes three different cell types: unigenomic spores, a compartmentalized mycelium (MI) and a multinucleated mycelium (substrate and aerial mycelium, MII). The importance of DNA methylation was already described in Streptomycetes, but its biological role remained unknown. The main objectives of this study were to analyze the pattern of cytosine methylation in Streptomyces coelicolor and to investigate the relationship between DNA cytosine methylation and morphological/physiological differentiation. Dot-blot analysis of genomic DNA using antibody anti-5-methylcytosine revealed that DNA methylation is modulated during hyphae differentiation. Specifically DNA cytosine methylation is higher at the MI stage than in the MII or spores. Cytosine methylome was investigated by bisulphite DNA sequencing showing that 30% of S. coelicolor genes contain a methylated motif in their upstream region. The biological effect of cytosine methylation was studied using 5-aza-2’-deoxycytidine (aza-dC), a hypomethylating agent. Phenotypic analyses of cultures treated with aza-dC demonstrated that they were impaired in germination, aerial mycelium formation and sporulation. In addition, they showed a strong reduction in antibiotic production. Overall, our results suggest a role for DNA cytosine methylation in morphological and physiological differentiation of S. coelicolor. Further experiments are ongoing to characterize the molecular mechanisms and pathways behind the observed phenotypes.
- Published
- 2015
41. Can DNA methylation contribute to explain morphological variability? The case of loggerhead turtle Caretta caretta
- Author
-
CARACAPPA, Santo, PISCIOTTA, Annalisa, ALDUINA, Rosa, ARCULEO, Marco, Persichetti, MF, Caracappa, G, Caracappa, S, Pisciotta, A, Persichetti, MF, Caracappa, G, Alduina, R, and Arculeo, M
- Subjects
Caretta, epigenetic, DNA methylation - Abstract
The loggerhead turtle Caretta caretta is a common and threatened sea turtle in the Mediterranean Sea. It is listed in the Red List of Threatened Species of the International Union for Conservation of Nature (IUCN 2012) and, although it is widely studied, little is currently known about the relationships between its phenotype and genetic variability. In the last few years some authors observed the presence of individuals, collected in different parts of the Mediterranean Sea, with a variable number of carapacial and plastron scutes. This variability, in some cases, makes difficult the sound identification of the species (Margaritoulis and Chiras, 2011; Turkozan et al. 2001; Oliver 2014). To date, no one has tried to correlate these morphological anomalies with the genetic diversity of the species, even if several studies indicate that, beyond genetic variation, natural phenotypic variations can be generated through a suite of epigenetic mechanisms. Here we present some results about the possible correlation among the variability of the number of carapacial and plastron scutes and cytosine DNA methylation. 12 specimens of Caretta caretta, collected from different parts of Sicily, were analysed. Morphological characters were used for the identification of the collected specimens according to the guidelines of Marquez (1990). Afterwards, all the individuals were subjected to genetic analysis by the sequencing of a fragment of 243 bp of the mitochondrial cytochrome b gene. Genomic DNA methylation level of cytosines was evaluated by dot blot analysis using antibodies against methylated cytosines. Four of the twelve individuals analyzed showed an unexpected number of carapacial and plastron scutes making difficult the immediate and sound identification of the specimens as C. caretta. Sequences of the mitochondrial cytochrome b gene of these four individuals did not show any difference with those obtained from the other eight loggerhead turtles and with those from Genbank, thus allowing us to assign the four individuals to the species Caretta caretta. Conversely, dot blot analysis of genomic DNA showed a reduced global level of methylated cytosines in these four “abnormal” individuals in respect to the other ones. We hypothesize that the variability of the number of scutes could be described as an epigenetic effect probably due to the environmental parameters experienced by the embryos during incubation. This finding could have implications in our understanding of the pathways of morphological evolution and diversification in the chelonians.
- Published
- 2015
42. Omics approaches to elucidate the molecular physiology of lantibiotc NAI-107 production in Microbispora ATCC-PTA-5024
- Author
-
T. Faddetta, G. Gallo, G. Renzone, E. Palazzotto, P. Monciardini, S. Arena, A. Giardina, R. Alduina, T. Weber, F. Sangiorgi, A. Russo, G. Spinelli, SOSIO, MARGHERITA, A. Scaloni, A. M. Puglia, Faddetta, T., Gallo, G., Renzone, G., Palazzotto, E., Monciardini, P., Arena, S., Giardina, A., Alduina, R., Weber, T., Sangiorgi, F., Russo, A., Spinelli, G., Sosio, M., Scaloni, A., and Puglia, A.
- Subjects
Microbispora, NAI-107, 2D-DIGE ,Settore BIO/19 - Microbiologia Generale - Published
- 2015
43. Streptomyces coelicolor: DNA methylation and differentiation
- Author
-
PISCIOTTA, Annalisa, ALDUINA, Rosa, Galardini, M, Mengoni, A, Manteca, A, Pisciotta, A, Galardini, M, Mengoni, A, Manteca, A, and Alduina, R
- Subjects
DNA methylation, Streptomyces, morphological and physiological differentiation ,Settore BIO/19 - Microbiologia Generale - Abstract
DNA cytosine methylation is an epigenetic modification regulating many biological processes in eukaryotes, including chromatin organization, genome maintenance and gene expression. The role of DNA cytosine methylation in prokaryotes has not been deeply investigated. In Escherichia coli it was recently demonstrated that cytosine methylation regulates gene expression during stationary phase [1] and that an induced state of cytosine hypermethylation leads to chromosomal DNA cleavage and cell death [2]. Streptomyces coelicolor is a mycelial soil microorganism, which exhibits a complex life cycle that includes three different cell types: unigenomic spores, a compartmentalized mycelium (MI) and a multinucleated mycelium (substrate and aerial mycelium, MII) [3]. The importance of DNA methylation was already described in Streptomycetes [4], but its biological role remains unknown. The main objectives of this study are to analyze cytosine methylation pattern of Streptomyces coelicolor M145 during growth in liquid and on solid media, and to investigate the relationship between DNA cytosine methylation and morphological/physiological differentiation. Cytosine methylation of total genomic DNA extracted from different developmental stages was investigated by dot-blot experiments using antibody anti-5-methylcytosine. Cytosine methylome was analyzed by BiSulphite sequencing. The biological effect of cytosine methylation was studied adding 5-aza-2’-deoxycytidine (aza-dC), a hypomethylating agent, to the cultures. Dot blot analysis revealed that the level of cytosine methylation changes during development (MI, MII and spores). Specifically, DNA methylation is higher at the MI stage than in the MII or spores. BiSulphite sequencing revealed that 30% of S. coelicolor genes contained a methylated motif in their upstream regions. Genes harbouring these motifs included genes related to differentiation (aerial mycelium formation and sporulation), genes involved in DNA repair/replication/condensation, as well as genes encoding proteins with unknown functions. Phenotypic analyses of cultures treated with aza-dC demonstrated that DNA methylation influences germination, aerial mycelium formation and sporulation on solid medium and antibiotic production both, on solid and in liquid medium. Overall, our preliminary results suggest a role for DNA cytosine methylation in morphological and physiological differentiation of S. coelicolor. Further experiments are ongoing to demonstrate the molecular mechanisms and pathways behind the observed phenotypes.
- Published
- 2014
44. Inorganic phosphate is a trigger factor for Microbispora sp. ATCC-PTA-5024 growth and NAI-107 production
- Author
-
Margherita Sosio, Rosa Alduina, Paolo Monciardini, Anna Maria Puglia, Anna Giardina, Giuseppe Gallo, Giardina, A, Alduina, R, Gallo, G, Monciardini, P, Sosio, M, and Puglia, AM
- Subjects
food.ingredient ,Phosphate ,Bioengineering ,Biology ,Applied Microbiology and Biotechnology ,Phosphates ,Microbiology ,chemistry.chemical_compound ,food ,Bacteriocins ,Biosynthesis ,Polyphosphate ,Humans ,Ribosomal Post-translationally modified Peptides (RiPPs) ,2. Zero hunger ,PhoP-PhoR ,Research ,Structural gene ,Biological activity ,Lantibiotics ,biology.organism_classification ,Actinobacteria ,Ribosomal Post-translationally modified Peptides (RiPPs), Phosphate, PhoP-PhoR, Polyphosphate ,Chemically defined medium ,Regulon ,chemistry ,Biochemistry ,Microbispora ,Bacteria ,Biotechnology - Abstract
Background NAI-107, produced by the actinomycete Microbispora sp. ATCC-PTA-5024, is a promising lantibiotic active against Gram-positive bacteria and currently in late preclinical-phase. Lantibiotics (lanthionine-containing antibiotics) are ribosomally synthesized and post-translationally modified peptides (RiPPs), encoded by structural genes as precursor peptides. The biosynthesis of biologically active compounds is developmentally controlled and it depends upon a variety of environmental stimuli and conditions. Inorganic phosphate (Pi) usually negatively regulates biologically-active molecule production in Actinomycetes, while it has been reported to have a positive control on lantibiotic production in Firmicutes strains. So far, no information is available concerning the Pi effect on lantibiotic biosynthesis in Actinomycetes. Results After having developed a suitable defined medium, Pi-limiting conditions were established and confirmed by quantitative analysis of polyphosphate accumulation and of expression of selected Pho regulon genes, involved in the Pi-limitation stress response. Then, the effect of Pi on Microbispora growth and NAI-107 biosynthesis was investigated in a defined medium containing increasing Pi amounts. Altogether, our analyses revealed that phosphate is necessary for growth and positively influences both growth and NAI-107 production up to a concentration of 5 mM. Higher Pi concentrations were not found to further stimulate Microbispora growth and NAI-107 production. Conclusion These results, on one hand, enlarge the knowledge on Microbispora physiology, and, on the other one, could be helpful to develop a robust and economically feasible production process of NAI-107 as a drug for human use. Electronic supplementary material The online version of this article (doi:10.1186/s12934-014-0133-0) contains supplementary material, which is available to authorized users.
- Published
- 2014
45. Metabolic pathways in Microbispora sp. ATCC-PTA 5024, producer of NAI-107 lantibiotic
- Author
-
T. Faddetta, G. Gallo, E. Palazzotto, G. Renzone, A. Giardina, R. Alduina, P. Monciardini, T. Weber, M. Sosio, R. Pozzi, E. Stegmann, A. Russo, F. Sangiorgi, A. Scaloni, A. M. Puglia, Faddetta, T., Gallo, G., Palazzotto, E., Renzone, G., Giardina, A., Alduina, R., Monciardini, P., Weber, T., Sosio, M., Pozzi, R., Stegmann, E., Russo, A., Sangiorgi, F., Scaloni, A., and Puglia, A.
- Subjects
Microbispora sp. ATCC-PTA 5024, lantibiotic,NAI-107, 2D-DIGE ,Settore BIO/19 - Microbiologia Generale - Published
- 2014
46. Artificial Chromosomes to Explore and to Exploit Biosynthetic Capabilities of Actinomycetes
- Author
-
Rosa Alduina, Giuseppe Gallo, Alduina, R, and Gallo, G
- Subjects
Heterologous expression ,DNA, Bacterial ,Health, Toxicology and Mutagenesis ,lcsh:Biotechnology ,Heterologous ,lcsh:Medicine ,Human artificial chromosome ,Review Article ,Settore BIO/19 - Microbiologia Generale ,Streptomyces ,Genome ,Microbial biotechnology ,DNA sequencing ,Secondary metabolite ,03 medical and health sciences ,lcsh:TP248.13-248.65 ,Genetics ,Chromosomes, Artificial ,Molecular Biology ,Gene ,030304 developmental biology ,Gene Library ,0303 health sciences ,biology ,030306 microbiology ,Actinomycete ,lcsh:R ,General Medicine ,biology.organism_classification ,Artificial chromosome ,Biosynthetic Pathways ,Actinobacteria ,Multigene Family ,Molecular Medicine ,Bacteria ,Biotechnology - Abstract
Actinomycetes are an important source of biologically active compounds, like antibiotics, antitumor agents, and immunosuppressors. Genome sequencing is revealing that this class of microorganisms has larger genomes relative to other bacteria and uses a considerable fraction of its coding capacity (5–10%) for the production of mostly cryptic secondary metabolites. To access actinomycetes biosynthetic capabilities or to improve the pharmacokinetic properties and production yields of these chemically complex compounds, genetic manipulation of the producer strains can be performed. Heterologous expression in amenable hosts can be useful to exploit and to explore the genetic potential of actinomycetes and not cultivable but interesting bacteria. Artificial chromosomes that can be stably integrated into theStreptomycesgenome were constructed and demonstrated to be effective for transferring entire biosynthetic gene clusters from intractable actinomycetes into more suitable hosts. In this paper, the construction of several shuttleEscherichia coli-Streptomycesartificial chromosomes is discussed together with old and new strategies applied to improve heterologous production of secondary metabolites.
- Published
- 2012
47. Two heterologously expressed Planobispora rosea proteins cooperatively induce Streptomyces lividans thiostrepton uptake and storage from the extracellular medium
- Author
-
Valentina Di Caro, Roderich D. Süssmuth, Elvira M. Gottardi, Anna Giardina, Anna Maria Puglia, Rosa Alduina, Giardina, A, Alduina, R, Gottardi, E, Di Caro, V, Süssmuth, RD, and Puglia, AM
- Subjects
Chromosomes, Artificial, Bacterial ,lcsh:QR1-502 ,Bioengineering ,Applied Microbiology and Biotechnology ,Thiostrepton ,lcsh:Microbiology ,Microbiology ,chemistry.chemical_compound ,Open Reading Frames ,Bacterial Proteins ,Actinomycetales ,ORFS ,Cloning, Molecular ,Streptomyces nogalater ,Planobispora rosea, Streptomyces lividans, heterologous expression ,biology ,Research ,Nogalamycin ,Streptomyces coelicolor ,biology.organism_classification ,Anti-Bacterial Agents ,Complementation ,Subcloning ,chemistry ,Streptomyces lividans ,Biotechnology - Abstract
Background A bacterial artificial chromosomal library of Planobispora rosea, a genetically intractable actinomycete strain, was constructed using Escherichia coli-Streptomyces artificial chromosome (ESAC) and screened for the presence of genes known to be involved in the biosynthesis of antibiotics. Results One clone with a 40 kb insert showed antimicrobial activity against Gram positive bacteria. Insert sequence analysis and subcloning experiments revealed that the bioactivity was due to a 3.5 kb DNA fragment containing two open reading frames. These orfs encode two proteins with high similarity to a putative membrane protein of Streptomyces coelicolor and to the nogalamycin resistance protein SnorO of Streptomyces nogalater, respectively. The role of these two Orfs is unknown in Planobispora. Disruption and complementation experiments revealed that both proteins are necessary for the antibacterial activity and chemical analysis demonstrated that the antibiotic activity was due to thiostrepton, antibiotic used as recombinant clone selection marker. Conclusion Two Planobispora rosea orfs are responsible for increasing intracellular amounts and storage of thiostrepton in Streptomyces lividans.
- Published
- 2010
48. Phosphate-controlled regulator for the biosynthesis of the dalbavancin precursor A40926
- Author
-
Luca Lo Piccolo, Davide D'Alia, Anna Maria Puglia, Rosa Alduina, Stefano Donadio, Nina Gunnarsson, Clelia Ferraro, Groningen Biomolecular Sciences and Biotechnology, ALDUINA, R, LO PICCOLO, L, D'ALIA, D, FERRARO, C, GUNNARSSON, N, DONADIO, S, and PUGLIA, AM
- Subjects
GENE-CLUSTER ,Transcription, Genetic ,Operon ,SP ATCC-39727 ,Microbiology ,Phosphates ,PROMOTERS ,chemistry.chemical_compound ,Biosynthesis ,STRR ,Gene cluster ,STREPTOMYCES-GRISEUS ,Gene Regulation ,TRANSCRIPTION ,Phosphate-Controlled Regulator ,Promoter Regions, Genetic ,Molecular Biology ,Gene ,Antibacterial agent ,biology ,IDENTIFICATION ,Gene Expression Regulation, Bacterial ,biology.organism_classification ,GLYCOPEPTIDE ANTIBIOTIC A40926 ,Glycopeptide ,Anti-Bacterial Agents ,Actinobacteria ,Response regulator ,chemistry ,Biochemistry ,Multigene Family ,DNA-BINDING PROTEIN ,PHOR-PHOP SYSTEM ,Teicoplanin ,Streptomyces griseus - Abstract
The actinomycete Nonomuraea sp. strain ATCC 39727 produces the glycopeptide A40926, the precursor of the novel antibiotic dalbavancin. Previous studies have shown that phosphate limitation results in enhanced A40926 production. The A40926 biosynthetic gene ( dbv ) cluster, which consists of 37 genes, encodes two putative regulators, Dbv3 and Dbv4, as well as the response regulator (Dbv6) and the sensor-kinase (Dbv22) of a putative two-component system. Reverse transcription-PCR (RT-PCR) and real-time RT-PCR analysis revealed that the dbv14 - dbv8 and the dbv30 - dbv35 operons, as well as dbv4 , were negatively influenced by phosphate. Dbv4 shows a putative helix-turn-helix DNA-binding motif and shares sequence similarity with StrR, the transcriptional activator of streptomycin biosynthesis in Streptomyces griseus . Dbv4 was expressed in Escherichia coli as an N-terminal His 6 -tagged protein. The purified protein bound the dbv14 and dbv30 upstream regions but not the region preceding dbv4 . Bbr, a Dbv4 ortholog from the gene cluster for the synthesis of the glycopeptide balhimycin, also bound to the dbv14 and dbv30 upstream regions, while Dbv4 bound appropriate regions from the balhimycin cluster. Our results provide new insights into the regulation of glycopeptide antibiotics, indicating that the phosphate-controlled regulator Dbv4 governs two key steps in A40926 biosynthesis: the biosynthesis of the nonproteinogenic amino acid 3,5-dihydroxyphenylglycine and critical tailoring reactions on the heptapeptide backbone.
- Published
- 2007
49. Comparison of two PCR methods for detection of Leptospira interrogans in formalin-fixed and paraffin-embedded tissues
- Author
-
Maria Vitale, Yarelys Zamora Martínez, Carmen Fernandez Molina, Aleco D'Andrea, Rosa Alduina, V. Monteverde, D'Andrea, A, Martinez, YZ, Alduina, R, Monteverde, V, Molina, CF, and Vitale, M
- Subjects
DNA, Bacterial ,Microbiology (medical) ,Serotype ,lcsh:Arctic medicine. Tropical medicine ,Tissue Fixation ,lcsh:RC955-962 ,lcsh:QR1-502 ,Kidney ,Settore BIO/19 - Microbiologia Generale ,Polymerase Chain Reaction ,lcsh:Microbiology ,law.invention ,23S ribosomal RNA ,law ,Leptospira ,Formaldehyde ,RNA, Ribosomal, 16S ,medicine ,diagnostics ,Animals ,FFPE tissue ,Lung ,Polymerase chain reaction ,Paraffin Embedding ,biology ,microbiology ,Ribosomal RNA ,biology.organism_classification ,16S ribosomal RNA ,medicine.disease ,Leptospirosis ,Molecular biology ,RNA, Ribosomal, 23S ,PCR ,Cattle ,Leptospira interrogans - Abstract
In this study we compared two polymerase chain reaction (PCR) methods using either 16S ribosomal RNA (rRNA) or 23S rRNA gene primers for the detection of different Leptospira interrogans serovars. The performance of these two methods was assessed using DNA extracted from bovine tissues previously inoculated with several bacterial suspensions. PCR was performed on the same tissues before and after the formalin-fixed, paraffin-embedding procedure (FFPE tissues). The 23S rDNA PCR detected all fresh and FFPE positive tissues while the 16S rDNA-based protocol detected primarily the positive fresh tissues. Both methods are specific for pathogenic L. interrogans. The 23S-based PCR method successfully detected Leptospira in four dubious cases of human leptospirosis from archival tissue specimens and one leptospirosis-positive canine specimen. A sensitive method for leptospirosis identification in FFPE tissues would be a useful tool to screen histological specimen archives and gain a better assessment of human leptospirosis prevalence, especially in tropical countries, where large outbreaks can occur following the rainy season.
50. Differential proteomic analysis highlights metabolic strategies associated with balhimycin production in Amycolatopsis balhimycina chemostat cultivations
- Author
-
Andrea Scaloni, Jette Thykaer, Rosa Alduina, Giovanni Renzone, Linda Bianco, Anna Maria Puglia, Anna Eliasson-Lantz, Giuseppe Gallo, Gallo, G, Alduina, R, Renzone, G, Thykaer, J, Bianco, L, Eliasson-Lantz, A, Scaloni, A, and Puglia, AM
- Subjects
Proteome ,medicine.drug_class ,lcsh:QR1-502 ,Bioengineering ,Chemostat ,Biology ,Glycopeptide antibiotic ,Proteomics ,Settore BIO/19 - Microbiologia Generale ,Applied Microbiology and Biotechnology ,lcsh:Microbiology ,03 medical and health sciences ,Bacterial Proteins ,Vancomycin ,antibiotic ,Actinomycetales ,medicine ,Electrophoresis, Gel, Two-Dimensional ,Balhimycin ,proteomic ,030304 developmental biology ,2. Zero hunger ,chemistry.chemical_classification ,0303 health sciences ,030306 microbiology ,Research ,Fatty Acids ,Carbon ,Anti-Bacterial Agents ,Metabolic pathway ,glycopeptide ,Enzyme ,Glucose ,chemistry ,Biochemistry ,Amycolatopsis balhimycina ,Protein Biosynthesis ,Fermentation ,Biotechnology - Abstract
Background Proteomics was recently used to reveal enzymes whose expression is associated with the production of the glycopeptide antibiotic balhimycin in Amycolatopsis balhimycina batch cultivations. Combining chemostat fermentation technology, where cells proliferate with constant parameters in a highly reproducible steady-state, and differential proteomics, the relationships between physiological status and metabolic pathways during antibiotic producing and non-producing conditions could be highlighted. Results Two minimal defined media, one with low Pi (0.6 mM; LP) and proficient glucose (12 g/l) concentrations and the other one with high Pi (1.8 mM) and limiting (6 g/l; LG) glucose concentrations, were developed to promote and repress antibiotic production, respectively, in A. balhimycina chemostat cultivations. Applying the same dilution rate (0.03 h-1), both LG and LP chemostat cultivations showed a stable steady-state where biomass production yield coefficients, calculated on glucose consumption, were 0.38 ± 0.02 and 0.33 ± 0.02 g/g (biomass dry weight/glucose), respectively. Notably, balhimycin was detected only in LP, where quantitative RT-PCR revealed upregulation of selected bal genes, devoted to balhimycin biosynthesis, and of phoP, phoR, pstS and phoD, known to be associated to Pi limitation stress response. 2D-Differential Gel Electrophoresis (DIGE) and protein identification, performed by mass spectrometry and computer-assisted 2 D reference-map http://www.unipa.it/ampuglia/Abal-proteome-maps matching, demonstrated a differential expression for proteins involved in many metabolic pathways or cellular processes, including central carbon and phosphate metabolism. Interestingly, proteins playing a key role in generation of primary metabolism intermediates and cofactors required for balhimycin biosynthesis were upregulated in LP. Finally, a bioinformatic approach showed PHO box-like regulatory elements in the upstream regions of nine differentially expressed genes, among which two were tested by electrophoresis mobility shift assays (EMSA). Conclusion In the two chemostat conditions, used to generate biomass for proteomic analysis, mycelia grew with the same rate and with similar glucose-biomass conversion efficiencies. Global gene expression analysis revealed a differential metabolic adaptation, highlighting strategies for energetic supply and biosynthesis of metabolic intermediates required for biomass production and, in LP, for balhimycin biosynthesis. These data, confirming a relationship between primary metabolism and antibiotic production, could be used to increase antibiotic yield both by rational genetic engineering and fermentation processes improvement.
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.