Abstract: In Santos, coast of São Paulo State, the evaluation of the microbiological quality of the beach water is performed weekly by the Companhia Ambiental do Estado de São Paulo (CETESB), the environmental agency of the São Paulo state government. However, the same attention is not given to the sand of the beaches. Sand represents a potential contamination vector and may constitute a reservoir of pathogenic microorganisms. This study aimed to evaluate the microbiological quality of sands from Santos beaches throughout the quantification of bacteria colonies from the groups of fecal coliforms (Escherichia coli) and Enterococcus, using the membrane filter technique. The analyses were executed monthly during 8 months and the results, expressed in bacteria colony forming units (CFU) per 100 grams of sand, vary from 40,000 to 2,700,000 for E. coli and from not detected to 95,000 for Enterococcus, being above the currently guideline values in national (3,800 CFU.100g-1) and international (100,000 CFU.100g-1) levels. It’s important to mention that these guideline values were based on results from microbiological analyses of sands collected from beaches that are distant from large urban areas. The contact with contaminated sand may cause diseases, provoking impacts on the population’s life quality. There is a need for further studies based on epidemiological evidences of exposure and risk analysis, in order to establish quality standards and public policies aimed to monitoring and managing the sanitary quality of sands from São Paulo’s coast beaches.

Palavras-Chave: sand; seas; microorganisms; quality control; environment; environmental impacts; public health; bacteria; bacterial diseases; epidemiology; brazil

LESCRECK, MARINA C.; PETRONI, RENATA G.G.; CORTEZ, FERNANDO S.; SANTOS, ALDO R.; COUTINHO, POLLYANNA O.; PUSCEDDU, FABIO H. Análise da qualidade sanitária da areia das praias de Santos, litoral do estado de São Paulo. Engenharia Sanitaria e Ambiental, v. 21, n. 4, p. 777-782, 2016. DOI: 10.1590/S1413-41522016149550. Disponível em: http://repositorio.ipen.br/handle/123456789/27213. Acesso em: $DATA.

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Abstract: Mygalin is a synthetic analog of polyamine spermidine isolated from spider hemocytes. Polyamines show potential therapeutic activity against a wide range of human diseases such as cancer and microbial infections. In this work, we analyzed the antibacterial and antitumoral activities of Mygalin silver nanoparticles synthesized by the photoreduction method. The formation and distribution of MygAgNPs were confirmed by UV-visible spectroscopy, zeta potential, and transmission electron microscopy. The obtained nanoparticles were mostly spherical with a particle size distribution in the range of ~ 10–60 nm. We have demonstrated that MygAgNPs increased the effectiveness of the native Mygalin by approximately 6400-fold. Cytotoxicity tests were performed, and it was possible to reach a concentration that was not toxic to healthy cells (NHI-3T3) and at the same time toxic to the tumor cell line (MCF-7). The obtained results suggest that this system shows potential enhanced antibacterial activity against Escherichia coli, DH5α and anticancer activity against MCF-7 cell line.

Palavras-Chave: drugs; neoplasms; antibiotics; spiders; silver; nanoparticles; cell cultures; drugs; amines; bacteria

COURROL, LILIA C.; ESPINOZA-CULUPÚ, ABRAHAM; SILVA JUNIOR, PEDRO I. da; GONÇALVES, KARINA de O.; SILVA, FLAVIA R. de O.; BORGES, MONAMARIS M. Antibacterial and antitumoral activities of the spider acylpolyamine Mygalin silver nanoparticles. BioNanoScience, v. 10, n. 2, p. 463-472, 2020. DOI: 10.1007/s12668-020-00738-w. Disponível em: http://repositorio.ipen.br/handle/123456789/31350. Acesso em: $DATA.

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Abstract: Background: Antimicrobial photodynamic therapy (APDT) has been broadly investigated as an alternative to treat localized infections, without leading to the selection of resistant microorganisms. Infectious stomatitis is a multifactorial disease frequently reported in captive snakes characterized by infection of the oral mucosa and surrounding tissues. In this study, we investigated methylene blue (MB)-mediated APDT to treat infectious stomatitis in snakes and verified the resistance phenotype and genotype before and after APDT. Methods: Three Boid snakes presented petechiae, edema and caseous material in their oral cavities. MB (0.01%) was applied on the lesions and after 5 min they were irradiated using a red laser (λ=660 nm), fluence of 280 J/ cm2, 8 J and 80 s per point, 100 mW, spot size 0.028 cm2 and fluence rate of 3.5 W/cm2. APDT was repeated once a week during 3 months. Samples of the lesions were collected to identify bacteria and antibiotic resistance profiles. To analyze the clonality of bacterial isolates before and after APDT, isolates were subjected to ERIC PCR analysis. Results: Snakes presented clinical improvement such as reduction of inflammatory signs and caseous material. Pseudomonas aeruginosa and Escherichia coli were present in all snakes; Klebsiella pneumoniae and Morganella morganii were also identified in some animals. We also observed that the oral microbiota was completely replaced following APDT. However, K. pneumoniae isolates before and after APDT were a single clone with 100% of genetic similarity that lost resistance phenotype for seven antibiotics of four classes. Conclusions: These results show that APDT can be used to treat infectious stomatitis in snakes.

Palavras-Chave: antimicrobial agents; therapy; infectious diseases; veterinary medicine; bacteria; snakes; plasmids

GREGO, KATHLEEN F.; CARVALHO, MARCELO P.N. de; CUNHA, MARCOS P.V.; KNOBL, TEREZINHA; POGLIANI, FABIO C.; CATAO-DIAS, JOSE L.; SANT ANNA, SAVIO S.; RIBEIRO, MARTHA S.; SELLERA, FABIO P. Antimicrobial photodynamic therapy for infectious stomatitis in snakes: clinical views and microbiological findings. Photodiagnosis and Photodynamic Therapy, v. 20, p. 196-200, 2017. DOI: 10.1016/j.pdpdt.2017.10.004. Disponível em: http://repositorio.ipen.br/handle/123456789/28489. Acesso em: $DATA.

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Abstract: The inexorable increase of antibiotic resistance occurring in different bacterial species is increasing the interest in developing new antimicrobial treatments that will be equally effective against multidrug-resistant strains and will not themselves induce resistance. One of these alternatives may be photodynamic inactivation (PDI), which uses a combination of nontoxic dyes, called photosensitizers (PS), excited by harmless visible light to generate reactive oxygen species (ROS) by type 1 (radical) and type 2 (singlet oxygen) pathways. In this study, we asked whether it was possible to improve the efficacy of PDI in vitro and in vivo by addition of the inert salt potassium iodide (KI) to a commonly investigated PS, the phenothiazinium dye methylene blue (MB). By adding KI, we observed a consistent increase of red light-mediated bacterial killing of Gram-positive and Gram-negative species in vitro and in vivo. In vivo, we also observed less bacterial recurrence in wounds in the days posttreatment. The mechanism of action is probably due to formation of reactive iodine species that are produced quickly with a short lifetime. This finding may have a relevant clinical impact by reducing the risk of amputation and, in some cases, the risk of death, leading to improvement in the care of patients affected by localized infections.

Palavras-Chave: bacteria; inactivation; photosensitivity; methylene blue; potassium iodides; in vivo; in vitro; antibiotics; drugs; light sources; therapy; reagents

VECCHIO, DANIELA; GUPTA, ASHEESH; HUANG, LIYI; LANDI, GIACOMO; AVCI, PINAR; RODAS, ANDREA; HAMBLIN, MICHAEL R. Bacterial photodynamic inactivation mediated by methylene blue and red light is enhanced by synergistic effect of potassium iodide. Antimicrobial Agents and Chemotherapy, v. 59, n. 9, p. 5203-5212, 2015. DOI: 10.1128/AAC.00019-15. Disponível em: http://repositorio.ipen.br/handle/123456789/31145. Acesso em: $DATA.

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Abstract: Reversed-phase high-performance liquid chromatography (RP-HPLC) has been used to analyze Interferon α-2 (IFN-α2) as a pure protein or as a pharmaceutical preparation: a method for analyzing periplasmic IFN-α2 directly in osmotic shock extract has, however, never been reported. This work describes an RP-HPLC methodology for the qualitative and quantitative analysis of human IFN-α2a and IFN-α2b directly in bacterial periplasmic extracts or in purified preparations. The analytical method has been set up and validated for accuracy, precision, linearity, sensitivity and specificity. A recovery test indicated an average bias of ∼1%, intra-day and inter-day quantitative determinations presented relative standard deviations always ≤ 5%, while the working sensitivity was of ∼0.3 μg of IFN-α2 (RSD =5%). The method proved to be suitable for detecting and quantifying also glycosylated and oxidized forms and N-methionylated IFN-α2 molecules, it was, however, not able to distinguish between IFN-α2a and IFN-α2b. This rapid methodology allows the application of RP-HPLC as a powerful tool to monitor the production yield and quality of IFN-α2 in osmotic shock fluids, right after, or even during the fermentation process.

Palavras-Chave: bacteria; interferon; high-performance liquid chromatography; escherichia coli; qualitative chemical analysis; quantitative chemical analysis; fluids

DIAS, PAULO V.S.; ARTHUSO, FERNANDA S.; OLIVEIRA, JOAO E.; SUZUKI, MIRIAM F.; SOUSA, JOSE M.; RIBELA, MARIA T.C.P.; BARTOLINI, PAOLO; SOARES, CARLOS R.J. Determination of recombinant Interferon-α2 in E. coli periplasmic extracts by reversed-phase high-performance liquid chromatography. Journal of Chromatography B, v. 1072, p. 193-198, 2018. DOI: 10.1016/j.jchromb.2017.11.023. Disponível em: http://repositorio.ipen.br/handle/123456789/28449. Acesso em: $DATA.

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Abstract: Background Peri-implantitis treatments are mainly based on protocols for teeth but have not shown favorable outcomes for implants. The potential role of titanium dissolution products in peri-implantitis necessitate the consideration of material properties in devising treatment protocols. We assessed implant cleaning interventions on (1) bacterial removal from Ti-bound biofilms, (2) Ti surface alterations and related Ti particle dissolution, and (3) cytocompatibility. Methods Acid-etched Ti discs were inoculated with human peri-implant plaque biofilms and mechanical antimicrobial interventions were applied on the Ti-bound biofilms for 30 seconds each: (1) rotary nylon brush; (2) Ti brush; (3) water-jet on high and (4) low, and compared to sterile, untreated and Chlorhexidine-treated controls. We assessed colony forming units (CFU) counts, biofilm removal, surface changes via scanning electron microscopy (SEM) and atomic force microscopy (AFM), and Ti dissolution via light microscopy and Inductively-coupled Mass Spectrometry (ICP-MS). Biological effects of Ti particles and surfaces changes were assessed using NIH/3T3 fibroblasts and MG-63 osteoblastic cell lines, respectively. Results Sequencing revealed that the human biofilm model supported a diverse biofilm including known peri-implant pathogens. WJ and Nylon brush were most effective in reducing CFU counts (P < 0.01 versus control), whereas Chlorhexidine was least effective; biofilm imaging results were confirmatory. Ti brushes led to visible streaks on the treated surfaces, reduced corrosion resistance and increased Ti dissolution over 30 days of material aging as compared to controls, which increase was amplified in the presence of bacteria (all P-val < 0.05). Ti particles exerted cytotoxic effects against fibroblasts, whereas surfaces altered by Ti brushes exhibited reduced osteoconductivity versus controls (P < 0.05). Conclusions Present findings support that mechanical treatment strategies selected for implant biofilm removal may lead to Ti dissolution. Ti dissolution should become an important consideration in the clinical selection of peri-implantitis treatments and a necessary criterion for the regulatory approval of instruments for implant hygiene.

Palavras-Chave: dentistry; implants; titanium; bacteria; corrosion; surface properties; antimicrobial agents

KOTSAKIS, GEORGIOS A.; BLACK, RACHEL; KUM, JASON; BERBEL, LARISSA; SADR, ALI; KAROUSSIS, IOANNIS; SIMOPOULOU, MARA; DAUBERT, DIANE. Effect of implant cleaning on titanium particle dissolution and cytocompatibility. Journal of Periodontology, v. 92, n. 4, p. 580-591, 2021. DOI: 10.1002/JPER.20-0186. Disponível em: http://repositorio.ipen.br/handle/123456789/32062. Acesso em: $DATA.

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