Search results for: Valeria Amoretti

Authors: Valeria Dilcheva, Svetlozara Petkova, Ivelin Vladov

Abstract:

Nemathodes of the genus Trichinella are zoonotic parasites with a cosmopolitan distribution. More than 100 species of mammals, birds and reptiles are involved in the natural cycle of this nematode. At present, T. spiralis, T. pseudospiralis, and T. britovi have been found in Bulgaria. The existence of natural wildlife and domestic reservoirs of Trichinella spp. can be a serious threat to human health. Three trichinella isolates caused human trichinella infection outbreaks from three regions of Sofia City Province were used for the research: sample No. 1 - Ratus norvegicus, sample No. 2 – domestic pig (Sus scrofa domestica), sample No. 3 - domestic pig (Sus scrofa domestica). Trichinella larvae of the studied species were isolated via digestive method (pepsin, hydrochloric acid, water) at 37ºC by standard procedure and were determined by gender (male and female) based on their morphological characteristics. As a reference trichinella species were used: T. spiralis, T. pseudospiralis, T. nativa and T. britovi. Single male and female larvae of the three isolates were crossed with single male and female larvae of the reference trichinella species as well as reciprocally. As a result of cross-breeding, offspring of muscular larvae with T. spiralis and T. britovi were obtained, while in experiments with T. pseudospiralis and T. nativa, trichinella larvae were not found in the laboratory mice. The results obtained in the control groups indicate that the trichinella larvae used from the isolates and the four trichinella species are infective. Also, the infective ability of the F1 offspring from the successful cross-breeding between isolates and reference species was investigated. Through the data obtained in the experiment was found that isolates No. 1 and No. 2 belong to the species T. spiralis, and isolate No. 3 belongs to the species T. britovi. The results were confirmed by PCR and real-time PCR analysis. Thus the presence and circulation of the species T. spiralis and T. britovi in Bulgaria was confirmed. Probably the rodents (rats) are involved in the distribution of T. spiralis in urban environment. The species T. britovi found in a domestic pig speaks of some contact with wild animals for which T. britovi is characteristic. The probable reason is that a large number of farmers in Bulgaria practice the free-range breeding of domestic pigs. Part of the farmers also used as food for domestic pigs waste products from the game (foxes, jackals, bears, wolves) and probably thus the infection was obtained. The distribution range of trichinella species in Bulgaria is not strictly outlined. It is believed that T. spiralis is most common in domestic animals and T. britovi and T. pseudospiralis are characteristic of wildlife. To answer the question whether wild and synanthropic animals are infected with the same or different trichinella species, which species predominate in nature and what their distribution among different hosts is, further research is required.

Keywords: cross-breeding, Sofia, trichinellosis, Trichinella britovi, Trichinella spiralis

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Authors: Valeria Panzetta, Ida Musella, Sabato Fusco, Paolo Antonio Netti

Abstract:

The study of the biophysics of living cells drew attention to the pivotal role of the cytoskeleton in many cell functions, such as mechanics, adhesion, proliferation, migration, differentiation and neoplastic transformation. In particular, during the complex process of malignant transformation and invasion cell cytoskeleton devolves from a rigid and organized structure to a more compliant state, which confers to the cancer cells a great ability to migrate and adapt to the extracellular environment. In order to better understand the malignant transformation process from a mechanical point of view, it is necessary to evaluate the direct crosstalk between the cells and their surrounding extracellular matrix (ECM) in a context which is close to in vivo conditions. In this study, human biopsy tissues of lung adenocarcinoma were analyzed in order to define their mechanical phenotype at cell and ECM level, by using particle tracking microrheology (PTM) technique. Polystyrene beads (500 nm) were introduced into the sample slice. The motion of beads was obtained by tracking their displacements across cell cytoskeleton and ECM structures and mean squared displacements (MSDs) were calculated from bead trajectories. It has been already demonstrated that the amplitude of MSD is inversely related to the mechanical properties of intracellular and extracellular microenvironment. For this reason, MSDs of particles introduced in cytoplasm and ECM of healthy and tumor tissues were compared. PTM analyses showed that cancerous transformation compromises mechanical integrity of cells and extracellular matrix. In particular, the MSD amplitudes in cells of adenocarcinoma were greater as compared to cells of normal tissues. The increased motion is probably associated to a less structured cytoskeleton and consequently to an increase of deformability of cells. Further, cancer transformation is also accompanied by extracellular matrix stiffening, as confirmed by the decrease of MSDs of matrix in tumor tissue, a process that promotes tumor proliferation and invasiveness, by activating typical oncogenic signaling pathways. In addition, a clear correlation between MSDs of cells and tumor grade was found. MSDs increase when tumor grade passes from 2 to 3, indicating that cells undergo to a trans-differentiation process during tumor progression. ECM stiffening is not dependent on tumor grade, but the tumor stage resulted to be strictly correlated with both cells and ECM mechanical properties. In fact, a greater stage is assigned to tumor spread to regional lymph nodes and characterized by an up-regulation of different ECM proteins, such as collagen I fibers. These results indicate that PTM can be used to get nanomechanical characterization at different scale levels in an interpretative and diagnostic context.

Keywords: cytoskeleton, extracellular matrix, mechanical properties, particle tracking microrheology, tumor

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Authors: Leto Leandra, Guaitini Caterina, Agosti Anna, Del Vecchio Lorenzo, Guarrasi Valeria, Cirlini Martina, Chiancone Benedetta

Abstract:

To the best of our knowledge, this study represents the first attempt to investigate the in vitro germination response of Rhus coriaria L. and its sprout chemical characterization. Rhus coriaria L., a species belonging to the Anacardiaceae family, is commonly called "sumac” and is cultivated, in different countries of the Mediterranean and the Middle East regions, to produce a spice with a sour taste, obtained from its dried and ground fruits. Moreover, since ancient times, many beneficial properties have been attributed to this plant that has been used, in the traditional medicine of several Asian countries, against various diseases, including liver and intestinal pathologies, ulcers, and various inflammatory states. In the recent past, sumac was cultivated in the Southern regions of Italy to treat leather, but its cultivation was abandoned, and currently, sumac plants grow spontaneously in marginal areas. Recently, in Italy, the interest in this species has been growing again, thanks to its numerous properties; thus, it becomes imperative to deepen the knowledge of this plant. In this study, in order to set up an efficient in vitro seed germination protocol, sumac seeds collected from spontaneous plants grown in Sicily, an island in the South of Italy, were, firstly, subjected to different treatments, scarification (mechanical, physical and chemical), cold stratification and imbibition, to break their physical and physiological dormancy, then, treated seeds were in vitro cultured on media with different gibberellic acid (GA3) concentrations. Results showed that, without any treatment, only 5% of in vitro sown seeds germinated, while the germination percentage increased up to 19% after the mechanical scarification. A further significative improvement of germination percentages was recorded after the physical scarification, with (40.5%) or without (36.5%) 8 weeks of cold stratification, especially when seeds were sown on gibberellin enriched cultured media. Vitro-derived sumac sprouts, at different developmental stages, were chemically characterized, in terms of polyphenol and tannin content, as well as for their antioxidant activity, to evaluate this matrix as a potential novel food or as a source of bioactive compounds. Results evidenced how more developed sumac sprouts and, above all, their leaves are a wealthy source of polyphenols (78.4 GAE/g SS) and tannins (21.9 mg GAE/g SS), with marked antioxidant activity. The outcomes of this study will be of support the nursery sector and sumac growers in obtaining a higher number of plants in a shorter time; moreover, the sprout chemical characterization will contribute to the process of considering this matrix as a new source of bioactive compounds and tannins to be used in food and non-food sectors.

Keywords: bioactive compounds, germination pre-treatments, rhus coriaria l., tissue culture

Procedia PDF Downloads 47

Authors: Leto Leandra, Guaitini Caterina, Agosti Anna, Del Vecchio Lorenzo, Guarrasi Valeria, Cirlini Martina, Chiancone Benedetta

Abstract:

To the best of our knowledge, this study represents the first attempt to investigate the in vitro germination response of Rhus coriaria L., and its sprout chemical characterization. Rhus coriaria L., a species belonging to the Anacardiaceae family, is commonly called "sumac" and is cultivated, in different countries of the Mediterranean and the Middle East regions, to produce a spice with a sour taste, obtained from its dried and ground fruits. Moreover, since ancient times, many beneficial properties have been attributed to this plant that has been used, in the traditional medicine of several Asian countries, against various diseases, including liver and intestinal pathologies, ulcers and various inflammatory states. In the recent past, sumac was cultivated in the Southern regions of Italy to treat leather, but its cultivation was abandoned, and currently, sumac plants grow spontaneously in marginal areas. Recently, in Italy, the interest in this species has been growing again, thanks to its numerous properties; thus, it becomes imperative to deepen the knowledge of this plant. In this study, in order to set up an efficient in vitro seed germination protocol, sumac seeds collected from spontaneous plants grown in Sicily, an island in the South of Italy, were, firstly, subjected to different treatments, scarification (mechanical, physical and chemical), cold stratification and imbibition, to break their physical and physiological dormancy, then, treated seeds were in vitro cultured on media with different gibberellic acid (GA3) concentrations. Results showed that, without any treatment, only 5% of in vitro sown seeds germinated, while the germination percentage increased up to 19% after the mechanical scarification. A further significative improvement of germination percentages was recorded after the physical scarification, with (40.5%) or without (36.5%) 8 weeks of cold stratification, especially when seeds were sown on gibberellin enriched cultured media. Vitro-derived sumac sprouts, at different developmental stages, were chemically characterized, in terms of polyphenol and tannin content, as well as for their antioxidant activity, to evaluate this matrix as a potential novel food or as a source of bioactive compounds. Results evidenced how more developed sumac sprouts and, above all, their leaves are a wealthy source of polyphenols (78.4 GAE/g SS) and tannins (21.9 mg GAE/g SS), with marked antioxidant activity. The outcomes of this study will be of support the nursery sector and sumac growers in obtaining a higher number of plants in a shorter time; moreover, the sprout chemical characterization will contribute to the process of considering this matrix as a new source of bioactive compounds and tannins to be used in food and non-food sectors.

Keywords: bioactive compounds, germination pre-treatments, rhus coriaria l., tissue culture

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Authors: Al Sayed Al Soudy, Valeria Maselli, Gianluca Polese, Anna Di Cosmo

Abstract:

Cephalopods are considered as a model organism with a rich behavioral repertoire. Sophisticated behaviors were widely studied and described in different species such as Octopus vulgaris, who has evolved the largest and more complex nervous system among invertebrates. In O. vulgaris, cognitive abilities in problem-solving tasks and learning abilities are associated with long-term memory and spatial memory, mediated by highly developed sensory organs. They are equipped with sophisticated eyes, able to discriminate colors even with a single photoreceptor type, vestibular system, ‘lateral line analogue’, primitive ‘hearing’ system and olfactory organs. They can recognize chemical cues either through direct contact with odors sources using suckers or by distance through the olfactory organs. Cephalopods are able to detect widespread waterborne molecules by the olfactory organs. However, many volatile odorant molecules are insoluble or have a very low solubility in water, and must be perceived by direct contact. O. vulgaris, equipped with many chemosensory neurons located in their suckers, exhibits a peculiar behavior that can be provocatively described as 'smell by touch'. The aim of this study is to establish the priority given to chemical vs. visual perception in food choice. Materials and methods: Three different types of food (anchovies, clams, and mussels) were used, and all sessions were recorded with a digital camera. During the acclimatization period, Octopuses were exposed to the three types of food to test their natural food preferences. Later, to verify if food preference is maintained, food was provided in transparent screw-jars with pierced lids to allow both visual and chemical recognition of the food inside. Subsequently, we tested alternatively octopuses with food in sealed transparent screw-jars and food in blind screw-jars with pierced lids. As a control, we used blind sealed jars with the same lid color to verify a random choice among food types. Results and discussion: During the acclimatization period, O. vulgaris shows a higher preference for anchovies (60%) followed by clams (30%), then mussels (10%). After acclimatization, using the transparent and pierced screw jars octopus’s food choices resulted in 50-50 between anchovies and clams, avoiding mussels. Later, guided by just visual sense, with transparent but not pierced jars, their food preferences resulted in 100% anchovies. With pierced but not transparent jars their food preference resulted in 100% anchovies as first food choice, the clams as a second food choice result (33.3%). With no possibility to select food, neither by vision nor by chemoreception, the results were 20% anchovies, 20% clams, and 60% mussels. We conclude that O. vulgaris uses both chemical and visual senses in an integrative way in food choice, but if we exclude one of them, it appears clear that its food preference relies on chemical sense more than on visual perception.

Keywords: food choice, Octopus vulgaris, olfaction, sensory organs, visual sense

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Authors: Valeria Cotza

Abstract:

School drop-out is a multifactorial phenomenon that in Italy concerns all those underage students who, at different school stages (up to 16 years old) or training (up to 18 years old), manifest educational difficulties from dropping out of compulsory education without obtaining a qualification to repetition rates and absenteeism. From the 1980s to the 2000s, there was a progressive attenuation of the economic and social model towards a multifactorial reading of the phenomenon, and the European Commission noted the importance of learning about the phenomenon through approaches able to integrate large-scale quantitative surveys with qualitative analyses. It is not a matter of identifying the contextual factors affecting the phenomenon but problematising them by means of systemic and comprehensive in-depth analysis. So, a privileged point of observation and field of intervention are those schools that propose alternative models of teaching and learning to the traditional ones, such as popular and second-chance schools. Alternative schools and interventions grew in these years in Europe as well as in the US and Latin America, working in the direction of greater equity to create the conditions (often absent in conventional schools) for everyone to achieve educational goals. Against extensive Anglo-Saxon and US literature on this topic, there is yet no unambiguous definition of alternative education, especially in Europe, where second-chance education has been most studied. There is little literature on a second chance in Italy and almost none on alternative education (with the exception of method schools, to which in Italy the concept of “alternative” is linked). This research aims to fill the gap by systematically surveying the alternative interventions in the area and beginning to explore some models of popular and second-chance schools and experiences through a mixed methods approach. So, the main research objectives concern the spread of alternative education in the Lombardy region, the main characteristics of these schools and interventions, and their effectiveness in terms of students’ well-being and school results. This paper seeks to answer the first point by presenting the preliminary results of the first phase of the project dedicated to mapping. Through the Google Forms platform, a questionnaire is being distributed to all schools in Lombardy and some schools in the rest of Italy to map the presence of alternative schools and interventions and their main characteristics. The distribution is also taking place thanks to the support of the Milan Territorial and Lombardy Regional School Offices. Moreover, other social realities outside the school system (such as cooperatives and cultural associations) can be questioned. The schools and other realities to be questioned outside Lombardy will also be identified with the support of INDIRE (Istituto Nazionale per Documentazione, Innovazione e Ricerca Educativa, “National Institute for Documentation, Innovation and Educational Research”) and based on existing literature and the indicators of “Futura” Plan of the PNRR (Piano Nazionale di Ripresa e Resilienza, “National Recovery and Resilience Plan”). Mapping will be crucial and functional for the subsequent qualitative and quantitative phase, which will make use of statistical analysis and constructivist grounded theory.

Keywords: school drop-out, alternative education, popular and second-chance schools, map

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Authors: Ntandazo Dlatu, Benjamin Longo-Mbenza, Andre Renzaho, Ruffin Appalata, Yolande Yvonne Valeria Matoumona Mavoungou, Mbenza Ben Longo, Kenneth Ekoru, Blaise Makoso, Gedeon Longo Longo

Abstract:

Background: The gap between complexities related to the integration of Tuberculosis /HIV control and evidence-based knowledge motivated the initiation of the study. Therefore, the objective of this study was to explore correlations between national TB management guidelines, multiple deprivation indexes, quantiles, components and levels of Tuberculosis control programme using mathematical modeling in rural O.R. Tambo District Health Facilities, South Africa. Methods: The study design used mixed secondary data analysis and cross-sectional analysis between 2009 and 2013 across O.R Tambo District, Eastern Cape, South Africa using univariate/ bivariate analysis, linear multiple regression models, and multivariate discriminant analysis. Health inequalities indicators and component of an impediment to the tuberculosis control programme were evaluated. Results: In total, 62 400 records for TB notification were analyzed for the period 2009-2013. There was a significant but negative between Financial Year Expenditure (r= -0.894; P= 0.041) Seropositive HIV status(r= -0.979; P= 0.004), Population Density (r = -0.881; P= 0.048) and the number of TB defaulter in all TB cases. It was shown unsuccessful control of TB management program through correlations between numbers of new PTB smear positive, TB defaulter new smear-positive, TB failure all TB, Pulmonary Tuberculosis case finding index and deprivation-concentration-dispersion index. It was shown successful TB program control through significant and negative associations between declining numbers of death in co-infection of HIV and TB, TB deaths all TB and SMIAD gradient/ deprivation-concentration-dispersion index. The multivariate linear model was summarized by unadjusted r of 96%, adjusted R2 of 95 %, Standard Error of estimate of 0.110, R2 changed of 0.959 and significance for variance change for P=0.004 to explain the prediction of TB defaulter in all TB with equation y= 8.558-0.979 x number of HIV seropositive. After adjusting for confounding factors (PTB case finding the index, TB defaulter new smear-positive, TB death in all TB, TB defaulter all TB, and TB failure in all TB). The HIV and TB death, as well as new PTB smear positive, were identified as the most important, significant, and independent indicator to discriminate most deprived deprivation index far from other deprivation quintiles 2-5 using discriminant analysis. Conclusion: Elimination of poverty such as overcrowding, lack of sanitation and environment of highest burden of HIV might end the TB threat in O.R Tambo District, Eastern Cape, South Africa. Furthermore, ongoing adequate budget comprehensive, holistic and collaborative initiative towards Sustainable Developmental Goals (SDGs) is necessary for complete elimination of TB in poor O.R Tambo District.

Keywords: tuberculosis, HIV/AIDS, success, failure, control program, health inequalities, South Africa

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Authors: Irene Carmagnola, Valeria Chiono, Sandra Pacharra, Jochen Salber, Sean McMahon, Chris Lovell, Pooja Basnett, Barbara Lukasiewicz, Ipsita Roy, Xiang Zhang, Gianluca Ciardelli

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Thrombosis and restenosis after stenting procedure can be prevented by promoting fast stent wall endothelialisation. It is well known that surface functionalisation with antifouling molecules combining with extracellular matrix proteins is a promising strategy to design biomimetic surfaces able to promote fast endothelialization. In particular, REDV has gained much attention for the ability to enhance rapid endothelialization due to its specific affinity with endothelial cells (ECs). In this work, a two-step plasma treatment was performed to polymerize a thin layer of acrylic acid, used to subsequently graft PEGylated-REDV and polyethylene glycol (PEG) at different molar ratio with the aim to selectively promote endothelial cell adhesion avoiding platelet activation. PEGylate-REDV was provided by Biomatik and it is formed by 6 PEG monomer repetitions (Chempep Inc.), with an NH2 terminal group. PEG polymers were purchased from Chempep Inc. with two different chain lengths: m-PEG6-NH2 (295.4 Da) with 6 monomer repetitions and m-PEG12-NH2 (559.7 Da) with 12 monomer repetitions. Plasma activation was obtained by operating at 50W power, 5 min of treatment and at an Ar flow rate of 20 sccm. Pure acrylic acid (99%, AAc) vapors were diluted in Ar (flow = 20 sccm) and polymerized by a pulsed plasma discharge applying a discharge RF power of 200 W, a duty cycle of 10% (on time = 10 ms, off time = 90 ms) for 10 min. After plasma treatment, samples were dipped into an 1-(3-dimethylaminopropyl)-3- ethylcarbodiimide (EDC)/N-hydroxysuccinimide (NHS) solution (ratio 4:1, pH 5.5) for 1 h at 4°C and subsequently dipped in PEGylate-REDV and PEGylate-REDV:PEG solutions at different molar ratio (100 μg/mL in PBS) for 20 h at room temperature. Surface modification was characterized through physico-chemical analyses and in vitro cell tests. PEGylated-REDV peptide and PEG were successfully bound to the carboxylic groups that are formed on the polymer surface after plasma reaction. FTIR-ATR spectroscopy, X -ray Photoelectron Spectroscopy (XPS) and contact angle measurement gave a clear indication of the presence of the grafted molecules. The use of PEG as a spacer allowed for an increase in wettability of the surface, and the effect was more evident by increasing the amount of PEG. Endothelial cells adhered and spread well on the surfaces functionalized with the REDV sequence. In conclusion, a selective coating able to promote a new endothelial cell layer on polymeric stent surface was developed. In particular, a thin AAc film was polymerised on the polymeric surface in order to expose –COOH groups, and PEGylate-REDV and PEG were successful grafted on the polymeric substrates. The REDV peptide demonstrated to encourage cell adhesion with a consequent, expected improvement of the hemocompatibility of these polymeric surfaces in vivo. Acknowledgements— This work was funded by the European Commission 7th Framework Programme under grant agreement number 604251- ReBioStent (Reinforced Bioresorbable Biomaterials for Therapeutic Drug Eluting Stents). The authors thank all the ReBioStent partners for their support in this work.

Keywords: endothelialisation, plasma treatment, stent, surface functionalisation

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Authors: Viviana Manzulli, Luigina Serrecchia, Adelia Donatiello, Valeria Rondinone, Sabine Zange, Alina Tscherne, Antonio Parisi, Antonio Fasanella

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Anthrax is a zoonotic disease that affects a wide range of animal species (primarily ruminant herbivores), and can be transmitted to humans through consumption or handling of contaminated animal products. The etiological agent B.anthracis is able to survive in unfavorable environmental conditions by forming endospore which remain viable in the soil for many decades. Furthermore, B.anthracis is considered as one of the most feared agents to be potentially misused as a biological weapon and the importance of the disease and its treatment in humans has been underscored before the bioterrorism events in the United States in 2001. Due to the often fatal outcome of human cases, antimicrobial susceptibility testing plays especially in the management of anthrax infections an important role. In Italy, animal anthrax is endemic (predominantly found in the southern regions and on islands) and is characterized by sporadic outbreaks occurring mainly during summer. Between 2012 and 2017 single human cases of cutaneous anthrax occurred. In this study, 90 diverse strains of B.anthracis, isolated in Italy from 2001 to 2017, were screened to their susceptibility to sixteen clinically relevant antimicrobial agents by using the broth microdilution method. B.anthracis strains selected for this study belong to the strain collection stored at the Anthrax Reference Institute of Italy located inside the Istituto Zooprofilattico Sperimentale of Puglia and Basilicata. The strains were isolated at different time points and places from various matrices (human, animal and environmental). All strains are a representative of over fifty distinct MLVA 31 genotypes. The following antibiotics were used for testing: gentamicin, ceftriaxone, streptomycin, penicillin G, clindamycin, chloramphenicol, vancomycin, linezolid, cefotaxime, tetracycline, erythromycin, rifampin, amoxicillin, ciprofloxacin, doxycycline and trimethoprim. A standard concentration of each antibiotic was prepared in a specific diluent, which were then twofold serial diluted. Therefore, each wells contained: bacterial suspension of 1–5x104 CFU/mL in Mueller-Hinton Broth (MHB), the antibiotic to be tested at known concentration and resazurin, an indicator of cell growth. After incubation overnight at 37°C, the wells were screened for color changes caused by the resazurin: a change from purple to pink/colorless indicated cell growth. The lowest concentration of antibiotic that prevented growth represented the minimal inhibitory concentration (MIC). This study suggests that B.anthracis remains susceptible in vitro to many antibiotics, in addition to doxycycline (MICs ≤ 0,03 µg/ml), ciprofloxacin (MICs ≤ 0,03 µg/ml) and penicillin G (MICs ≤ 0,06 µg/ml), recommend by CDC for the treatment of human cases and for prophylactic use after exposure to the spores. In fact, the good activity of gentamicin (MICs ≤ 0,25 µg/ml), streptomycin (MICs ≤ 1 µg/ml), clindamycin (MICs ≤ 0,125 µg/ml), chloramphenicol(MICs ≤ 4 µg/ml), vancomycin (MICs ≤ 2 µg/ml), linezolid (MICs ≤ 2 µg/ml), tetracycline (MICs ≤ 0,125 µg/ml), erythromycin (MICs ≤ 0,25 µg/ml), rifampin (MICs ≤ 0,25 µg/ml), amoxicillin (MICs ≤ 0,06 µg/ml), towards all tested B.anthracis strains demonstrates an appropriate alternative choice for prophylaxis and/or treatment. All tested B.anthracis strains showed intermediate susceptibility to the cephalosporins (MICs ≥ 16 µg/ml) and resistance to trimethoprim (MICs ≥ 128 µg/ml).

Keywords: Bacillus anthracis, antibiotic susceptibility, treatment, minimum inhibitory concentration

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