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O objetivo do trabalho foi analisar os microrganismos presentes em aparelhos de academia e a eficiência da desinfecção com álcool 70%. Foi realizada uma pesquisa experimental, descritiva e quantitativa. A coleta foi realizada em superfícies de aparelhos/equipamentos de academia de uma instituição de ensino privada, antes e após desinfecção, em seguida as colônias crescidas foram identificadas pelo teste de coloração de Gram, prova de catalase e crescimento em Ágar manitol, e realizado o teste de susceptibilidade aos antibióticos. As placas de cultura apresentaram crescimento de colônias bacteriológicas antes da limpeza, e também foi possível observar crescimento em algumas placas após a desinfecção, ainda que em menor expressividade. Na identificação, houve predominância de Staphylococcus aureus, que apresentou boa sensibilidade ao norfloxacino (73%) e a eritromicina (59%), e resistência à penicilina (100%) e ertapenem (67%). Todas as superfícies dos instrumentos estudados estavam contaminadas e, apesar da eficácia do álcool 70%, são necessárias formas adicionais de prevenção.

O objetivo do trabalho foi avaliar a eficiência de agentes desinfetantes (detergente neutro, álcool á 70% e hipoclorito de sódio) em diferentes superfícies laboratorial (granito, mármore e inox) e sobre microrganismos (Staphylococcus aureus, Escherichia coli e Candida albicans). Para tal, foi realizado uma pesquisa experimental em superfícies de trabalho de um laboratório de análises clínica, os microrganismos foram cultivados e com auxílio de swab foram transferidos para as bancadas e após o crescimento, contadas as colônias e em seguida feito a antissepsia com as soluções em testes e conferidas as colônias. A partir das análises, o potencial microbicida do detergente neutro e do álcool á 70% contra os microrganismos é observável nas superfícies de granito, mármore e inox, o hipoclorito de sódio foi a solução que apresentou a mais baixa eficácia dos desinfetantes. Em relação as superfícies, o granito, foi o que teve o maior número de colônias e teve a menor ação das soluções. As soluções desinfetantes, são essenciais para a antissepsia de superfície, mas, algumas substâncias não apresentar uma eficiência adequada sobre todos os tipos de materiais usados em superfícies e microrganismos.

Cases of human infections related to species of the genus Corynebacterium spp. have increased gradually both in industrialized and developing countries. Species of the genus, often multidrug-resistant, has been cited as a pathogen of nosocomial infections associated with sepsis, endocarditis, infections of surgical wounds, prostheses, and infections related to the central venous catheter. Medicinal plants represent the oldest "weapons" used by humans for the treatment of all types of diseases, that is, the use of plants in the diseases prevention and/or healing is a habit that has always existed in human history. In this sense, this review aimed to research articles about the antibacterial action of medicinal plants against Corynebacterium species. The methodology used was bibliographic survey in the main scientific research databases. Seventy-one species of medicinal plants showed antibacterial activity against the genus Corynebacterium spp., in addition to the product havan samagri, which is composed of a mixture of 54 plant species; among them, eleven plants did not show any inhibitory activity for different species of that genus. Thus, there is a clear relevance in conducting new research in the area due to the possibility of validating popular information and the development of new antimicrobial drugs. Keywords: Medicinal Plants. Antibacterial Activity. Corynebacterium spp. Resumo Casos de infecções humanas relacionadas às espécies do gênero Corynebacterium têm sido relatados, tanto em países industrializados quanto em desenvolvimento. Espécies do gênero, muitas vezes, multirresistentes aos antimicrobianos, são citadas como agentes de quadros como sepse, endocardite, infecções de feridas cirúrgicas, próteses e infecções relacionadas ao cateter venoso central. As plantas medicinais representam as mais antigas “armas” empregadas pelo homem no tratamento de enfermidades de todos os tipos, ou seja, a utilização de plantas na prevenção e/ou na cura de doenças é um hábito que sempre existiu na história da humanidade. Nesse sentido, esta revisão teve como objetivo pesquisar artigos sobre plantas medicinais com atividade antibacteriana contra corinebactérias. A metodologia utilizada foi o levantamento bibliográfico nos principais bancos de dados de pesquisas científicas. Setenta e uma espécies de plantas medicinais, além do produto havan samagri, que é composto por uma mistura de 54 espécies vegetais, demonstraram atividade antibacteriana contra o gênero Corynebacterium. Onze plantas estudadas não apresentaram nenhuma atividade inibitória para diferentes espécies de corinebactérias. Em função da possibilidade de validar as informações populares e o desenvolvimento de novas drogas antimicrobianas fica clara a relevância da realização de novas pesquisas na área. Palavras-chave: Plantas Medicinais. Atividade Antibacteriana. Corynebacterium spp.

Profissionais da área de saúde utilizam luvas de procedimentos como equipamento de proteção individual (EPI) devido ao contato com material biológico, micro-organismos e agentes químicos. A literatura relata que a passagem de micro-organismos através das luvas é facilitada por pequenos defeitos no EPI, destacando a importância da higienização das mãos após sua retirada. A avaliação da eficácia e integridade das luvas é de suma importância, pois analisa se o EPI fornece segurança aos usuários envolvidos em atividades que oferecem riscos à saúde. Assim, o objetivo deste estudo foi verificar a passagem de Escherichia coli através de luvas de procedimentos antes de sua utilização. O presente estudo analisou 60 luvas que foram preenchidas com meio de cultura em caldo inoculado com a bactéria, amarradas pelo pulso e suspensas de modo que não tocassem em nenhuma superfície. Após 30 minutos, swabs estéreis embebidos em caldo foram passados em diferentes regiões e semeados em placas contendo meio de cultura. Em 2 luvas, observou-se o crescimento de colônias características do micro-organismo utilizado, cuja confirmação ocorreu pela microscopia após coloração pelo método de Gram. A visualização a vista desarmada não permitiu a detecção de perfurações, mesmo após leve compressão das luvas. Estes resultados indicam que pode haver microporos imperceptíveis nas luvas de procedimentos comercializadas, permitindo a passagem de micro-organismos e consequente contaminação do profissional e/ou do paciente. Palavras-chave: Biossegurança. Controle de Infecção. Permeabilidade de Luvas. Risco Ocupacional. Saúde do Trabalhador. Abstract Health professionals use procedure gloves as personal protective equipment (PPE) due to contact with biological material, microorganisms and chemical agents. The literature reports that the passage of microorganisms through gloves is facilitated by small defects in the PPE and highlights the importance of hand hygiene after its removal. Assessing the effectiveness and integrity of gloves is of paramount importance, as it analyzes whether the PPE provides safety to users involved in activities that pose health hazards. The aim of this study was to verify the passage of the Escherichia coli through procedure gloves before its use. Sixty gloves used to procedures were filled with tryptic soy broth inoculated with the bacteria, tied and suspended not touching any surface. After 30 minutes, sterile swab previously soaked in tryptic soy broth were passed in different regions and plated on MacConkey agar. After incubation period, was observed the presence of characteristic colonies of E. coli in two gloves, confirmed by Gram stain method. Naked eye examination did not detect perforations after slight compression of the gloves. The results show that procedure gloves can present micropores that allow the passage of microorganisms and consequent contamination of the professional or the patient. Keywords: Biosafety. Gloves Permeability. Infection Control. Occupational Risks. Occupational Health.

Objetivo do estudo foi avaliar as atividades antioxidante, antibacteriana e antibiofilme contra Corynebacterium ulcerans e toxicidade do Extrato Bruto Hidroalcóolico (EBH) de Stryphnodendron coriaceum Benth (barbatimão). As folhas de S. coriaceum foram submetidas a maceração com álcool a 70%. Avaliou-se a atividade antioxidante pelos métodos do 1,1-difenil-2-picrilhidrazil (DPPH) e redução do complexo de fosfomolibdênio. A atividade antibacteriana foi medidia por microdiluição (1:2) para as Concentrações Inibitória Mínima (CIM) e Bactericida Mínima (CBM) e a inibição da formação de biofilme em cinco isolados clínicos e um amostra padrão de C. ulcerans. O estudo da toxicidade foi realizado por testes de hemólise, alelopatia e Tenebrio molitor. O EBH apresentou capacidade antioxidantes, capturando 50,7% do DPPH na concentração de 0,37µg/mL e reduzindo 21,99% do fosfomolibdênio. A CIM observada foi de 500 µg/mL para as amostras, exceto a amostra 809, e o EBH não apresentou aspecto bactericida (CBM). A amostra CDC KC 279 diminuiu a formação do biofilme com interferência do EBH, enquanto, a amostra 2649 sofreu aumento (estatisticamente significativa). A concentração eficiente para hemolisar 50% das hemácias foi de 460,6±0,03881µg/mL. No teste alelopático, as sementes apresentaram baixa germinação e não houve diminuição na velocidade de crescimento das radículas em contato com o EBH. A taxa média de sobrevivência de T. molitor foi 60% no grupo da maior concentração, sem significância estatística (p=0,752) com o controle, essas características sugerem baixa toxicidade. A espécie vegetal em estudo, apresenta propriedades farmacológicas importantes, que pode levar a formulação de produtos terapêuticos, estudos complementares são necessários.

Cepas multirresistentes de Corynebacterium ulcerans destacam a necessidade de pesquisas em novos medicamentos e o uso farmacologico de oleos essenciais (OE) pode ser uma alternativa terapeutica. Assim, este estudo teve como objetivo avaliar a atividade antibacteriana e o efeito da concentracao subinibidora de AZEO na morfologia bacteriana e na formacao de biofilme na superficie do poliestireno por C. ulcerans utilizados em caes. O AZEO foi usado por destilacao a vapor das folhas da planta. Os testes de inibicao de fosfomolibdenio e hemolise foram realizados para analisar, respectivamente, uma atividade antioxidante e toxicidade. Foram selecionados como inibidores e bactericidas minimos (MIC e MBC) do AZEO em C. ulcerans e a partir desses resultados foram testados subinibidos dos testes do AZEO. A inatividade do complexo fosfomolibdenio pelo AZEO foi menor que 1% e a capacidade hemolitica foi baixa. Os resultados do estudo antimicrobiano mostram que o AZEO inibiu o crescimento de fundos como cepas microbianas testadas. Os filamentos bacterianos foram observados na presenca do AZEO , bem como uma bacteria de controle automatico. Houve diferenca significativa na inibicao do biofilme. Sugerimos que A. zerumbet pode usar uma terapia alternativa para controlar infeccoes bacterianas induzidas por C. ulcerans .

O objetivo desse trabalho foi avaliar os parasitas e Escherichia coli em alfaces (Lactuca sativa Linnaeus) comercializadas em feiras livres em São José de Ribamar-MA. Foram coletadas 30 amostras, na análise parasitológica utilizou-se o método de sedimentação/centrifugação e a microbiológica o kit ColiTest® e o meio ágar eosina azul de metileno para contagem de Escherichia coli. Todas as amostras continham algum tipo de protozoário e/ou helminto, sendo os protozoários Balantidium coli mais ocorrente na feira A e Entamoeba coli na B. Sobre os aspectos microbiológicos, todas as amostras apresentaram coliformes totais e Escherichia coli na amostra do vendedor 15 da feira A (3,87 x 102 UFC/mL) e o vendedor 4 da feira B (2,70 x 102 UFC/mL) foram que mais apresentaram contaminação por esse termololerante. As alfaces comercializadas encontram-se com restrição no consumo devido as condições higiênicas e sanitárias, assim, compreender os aspectos associados à contaminação dessas hortaliças é importante.

Biofilm-related infections are considered a major cause of morbidity and mortality in hospital environments. Biofilms allow microorganisms to exchange genetic material and to become persistent colonizers and/or multiresistant to antibiotics. Corynebacterium pseudodiphtheriticum (CPS), a commensal bacterium that colonizes skin and mucosal sites has become progressively multiresistant and responsible for severe nosocomial infections. However, virulence factors of this emergent pathogen remain unclear. Herein, we report the adhesive properties and biofilm formation on hydrophilic (glass) and hydrophobic (plastic) abiotic surfaces by CPS strains isolated from patients with localized (ATCC10700/Pharyngitis) and systemic (HHC1507/Bacteremia) infections. Adherence to polystyrene attributed to hydrophobic interactions between bacterial cells and this negatively charged surface indicated the involvement of cell surface hydrophobicity in the initial stage of biofilm formation. Attached microorganisms multiplied and formed microcolonies that accumulated as multilayered cell clusters, a step that involved intercellular adhesion and synthesis of extracellular matrix molecules. Further growth led to the formation of dense bacterial aggregates embedded in the exopolymeric matrix surrounded by voids, typical of mature biofilms. Data also showed CPS recognizing human fibrinogen (Fbg) and fibronectin (Fn) and involvement of these sera components in formation of "conditioning films". These findings suggested that biofilm formation may be associated with the expression of different adhesins. CPS may form biofilms in vivo possibly by an adherent biofilm mode of growth in vitro currently demonstrated on hydrophilic and hydrophobic abiotic surfaces. The affinity to Fbg and Fn and the biofilm-forming ability may contribute to the establishment and dissemination of infection caused by CPS.

Corynebacterium diphtheriae, the aetiologic agent of diphtheria, also represents a global medical challenge because of the existence of invasive strains as causative agents of systemic infections. Although tellurite (TeO32-) is toxic to most microorganisms, TeO32--resistant bacteria, including C. diphtheriae, exist in nature. The presence of TeO32--resistance (TeR) determinants in pathogenic bacteria might provide selective advantages in the natural environment. In the present study, we investigated the role of the putative TeR determinant (CDCE8392_813gene) in the virulence attributes of diphtheria bacilli. The disruption of CDCE8392_0813 gene expression in the LDCIC-L1 mutant increased susceptibility to TeO32- and reactive oxygen species (hydrogen peroxide), but not to other antimicrobial agents. The LDCIC-L1 mutant also showed a decrease in both the lethality of Caenorhabditis elegansand the survival inside of human epithelial cells compared to wild-type strain. Conversely, the haemagglutinating activity and adherence to and formation of biofilms on different abiotic surfaces were not regulated through the CDCE8392_0813 gene. In conclusion, the CDCE8392_813 gene contributes to the TeR and pathogenic potential of C. diphtheriae.

O presente trabalho teve como objetivo a realização de uma prospecção tecnológica sobre plantas medicinais com atividade antibacteriana. Foram utilizadas as bases de artigos científicos Science Direct, Web of Science e SCOPUS. Para teses e dissertações, o banco de Teses da CAPES, e para patentes, as bases do INPI, WIPO e EPO. A pesquisa foi realizada em julho de 2014, caracterizando um estudo documental. Foi possível observar um quantitativo superior de artigos científicos em relação as patentes sobre o tema abordado. Na base da WIPO, os descritores que mais almejaram o objetivo da pesquisa, foram encontrados 38 patentes, o país com maior número de depósito de patentes foi República da Coreia com 27, onde 32 estão classificadas na Seção A (necessidades humanas) e o ano que ocorreu mais depósito foi 2005, com 7 patentes. Há necessidade de mais investimentos em pesquisas e incentivos na elaboração de produtos terapêuticos.

Corynebacterium ulcerans é um patógeno emergente em vários países, inclusive no Brasil, e está envolvido em diversos quadros clínicos, incluindo a difteria. O estudo da atividade antibacteriana de óleos essenciais (OEs), extraídos de plantas de interesse farmacológico, na busca de um antimicrobiano ideal tem sido bastante difundido. O objetivo desta pesquisa foi avaliar o potencial antioxidante e a atividade antibacteriana do OE obtido da canela. A partir das folhas da planta, coletadas no município de São Luís-MA, extraiu-se o óleo por hidrodestilação por arraste a vapor. Para avaliação da ação antioxidante do OE em diferentes concentrações, utilizou-se o método de quelação do íon ferroso. A fim de determinar a atividade antibacteriana, testes de difusão (disco e poço) em ágar e de determinação das concentrações inibitória mínima (CIM) por microdiluição e bactericida mínima (CBM) foram realizados com seis isolados clínicos e uma amostra padrão de C. ulcerans. A capacidade do OE para quelar 50% do íon ferroso (CE50) foi de 464,3±0,1726 µg/mL. Nos testes de difusão em ágar, não foi observada a formação de halos de inibição bacteriana com as concentrações do OE utilizadas. Porém, foi possível determinar a CIM do OE para todas as amostras, sendo a menor de 3,90 µg/ml. Não se encontrou a CBM com as concentrações avaliadas. O OE de canela demonstrou ação antioxidante e antibacteriana contra C. ulcerans, sugerindo seu potencial para uso terapêutico.PALAVRAS-CHAVE: corinebactérias; planta medicinal; antimicrobiano

Although Corynebacterium diphtheriae has been classically described as an exclusively extracellular pathogen, there is growing evidence that it may be internalized by epithelial cells. The aim of the present report was to investigate the nature and involvement of the surface-exposed non-fimbrial 67-72kDa proteins (67-72p), previously characterized as adhesin/hemagglutinin, in C.diphtheriae internalization by HEp-2 cells. Transmission electron microscopy and bacterial internalization inhibition assays indicated the role of 67-72p as invasin for strains of varied sources. Cytoskeletal changes with accumulation of polymerized actin in HEp-2 cells beneath adherent 67-72p-adsorbed microspheres were observed by the Fluorescent actin staining test. Trypan blue staining method and Methylthiazole tetrazolium reduction assay showed a significant decrease in viability of HEp-2 cells treated with 67-72p. Morphological changes in HEp-2 cells observed after treatment with 67-72p included vacuolization, nuclear fragmentation and the formation of apoptotic bodies. Flow cytometry revealed an apoptotic volume decrease in HEp-2 cells treated with 67-72p. Moreover, a double-staining assay using Propidium Iodide/Annexin V gave information about the numbers of vital vs. early apoptotic cells and late apoptotic or secondary necrotic cells. The comparative analysis of MALDI-TOF MS experiments with the probes provided for 67-72p CDC-E8392 with an in silico proteome deduced from the complete genome sequence of C.diphtheriae identified with significant scores 67-72p as the protein DIP0733. In conclusion, DIP0733 (67-72p) may be directly implicated in bacterial invasion and apoptosis of epithelial cells in the early stages of diphtheria and C.diphtheriae invasive infection. Copyright 2011 Elsevier Ltd. All rights reserved.

During the last decade the majority of diphtheria cases in Europe had Corynebacterium ulcerans as the etiologic agent with dogs and cats as the reservoir hosts. However, little has been documented about the virulence factors of this zoonotic pathogen. To set up an in vivo experimental C. ulcerans infection model, conventional Swiss Webster mice were intravenously infected with different doses (from 1 × 10(7) to 5 × 10(9) bacteria per mouse) of C. ulcerans strains, namely 809 (from human lower respiratory tract), BR-AD22 (from asymptomatic dog nares) and CDC-KC279. Mortality rates were demonstrated by LD(50) values ranging from 1.9 × 10(8) to 1.3 × 10(9). Viable bacteria were recovered from blood, kidneys, liver, spleen and joints. For CDC-KC279 and 809 strains (2 × 10(8)mL(-1)) approximately 85% and 72% of animals with articular lesions were observed, respectively; BR-AD22-infected mice showed no signs of arthritis. CDC-KC279 and 809 strains exhibited higher arthritogenic potential when compared to the homologous toxigenic (ATCC27012) and non-toxigenic (ATCC27010) strains of Corynebacterium diphtheriae. A high number of affected joints and arthritis index in addition to the histopathological features, including subcutaneous edema, inflammatory infiltrate, damage to bone tissue and synoviocyte hypertrophy, indicated a strain-dependent ability of C. ulcerans strains to cause severe polyarthritis. A correlation between the arthritis index and systemic levels of IL-6 and TNF-α was observed for C. ulcerans strains, with the exception of the non-arthritogenic BR-AD22 strain. In conclusion, C. ulcerans revealed a strain-dependent arthritogenic potential independent of DNAse, PLD and diphtheria toxin production.

As interactions between bacteria and macrophages dictate the outcome of most infectious diseases, analyses of molecular mechanisms of non-opsonic phagocytosis should lead to new approaches for the prevention of diphtheria and systemic Corynebacterium diphtheriae infections. The present study aimed to evaluate human macrophage-bacteria interactions in the absence of opsonin antibodies and the influence of the tox gene on this process. Homologous C. diphtheriae tox+ and tox- strains were evaluated for adhesion, entering and survival within U-937 human macrophages at different incubation periods. Higher numbers of viable bacteria associated with and internalized by macrophages were demonstrated for the tox+ strain. However, viable intracellular bacteria were detected at T-24 hr only for the tox- strain. Cytoskeletal inhibitors, cytochalasin E, genistein and colchicine, inhibited intracellular viability of both strains at different levels. Bacterial replication was evidenced at T-24 hr in supernatants of monolayers infected with the tox- strain. Host cell death and nuclear alterations were evidenced by the Trypan blue exclusion assay and DAPI fluorescence microscopy. ELISA of histone-associated DNA fragments allowed detection of apoptosis and necrosis induced by tox+ and tox- strains at T-1 hr and T-3 hr. In conclusion, human macrophages in the absence of opsonins may not be promptly effective at killing diphtheria bacilli. The presence of the tox gene influences the susceptibility of C. diphtheriae to human macrophages and the outcome of non-opsonic phagocytosis. C. diphtheriae strains exhibit strategies to survive within macrophages and to exert apoptosis and necrosis in human phagocytic cells, independent of the tox gene.

Corynebacterium diphtheriae still represents a global medical challenge, particularly due to the significant number of individuals susceptible to diphtheria and the emergence of non-toxigenic strains as the causative agents of invasive infections. In this study, we characterized the clinical and microbiological features of what we believe to be the first case of C. diphtheriae infection of a percutaneous nephrostomy catheter insertion site in an elderly patient with a fatal bladder cancer. Moreover, we demonstrated the potential role of adherence, biofilm formation and fibrin deposition traits in C. diphtheriae from the catheter-related infection. Non-toxigenic C. diphtheriae isolated from the purulent discharge (named strain BR-CAT5003748) was identified by the API Coryne system (code 1 010 324) and a multiplex PCR for detection of dtxR and tox genes. Strain BR-CAT5003748 showed resistance to oxacillin, ceftazidime and ciprofloxacin. In experiments performed in vitro, the catheter isolate was classified as moderately hydrophobic and as moderately adherent to polystyrene surfaces. Glass provided a more effective surface for biofilm formation than polystyrene. Micro-organisms adhered to (>1.5×106 c.f.u.) and multiplied on surfaces of polyurethane catheters. Microcolony formation (a hallmark of biofilm formation) and amorphous accretions were observed by scanning electron microscopy on both external and luminal catheter surfaces. Micro-organisms yielded simultaneous expression of localized adherence-like and aggregative-like (LAL/AAL) adherence patterns to HEp-2 cells. Interestingly, the coagulase tube test resulted in the formation of a thin layer of fibrin embedded in rabbit plasma by the non-toxigenic BR-CAT5003748 strain. In conclusion, C. diphtheriae should be recognized as a potential cause of catheter-related infections in at-risk populations such as elderly and cancer patients. LAL/AAL strains may be associated with virulence traits that enable C. diphtheriae to effectively produce biofilms on catheter surfaces. Biofilm formation and fibrin deposition could have contributed to the persistence of C. diphtheriae at the infected insertion site and the obstruction of the nephrostomy catheter.

SUMMARYWe describe microbiological, clinical and epidemiological aspects of a diphtheria outbreak that occurred in Maranhão, Brazil. The majority of the 27 confirmed cases occurred in partially (n = 16) or completely (n = 10) immunized children (n = 26). Clinical signs and characteristic symptoms of diphtheria such as cervical lymphadenopathy and pseudomembrane formation were absent in 48% and 7% of the cases, respectively. Complications such as paralysis of lower limbs were observed. Three cases resulted in death, two of them in completely immunized children. Microbiological analysis identified the isolates asCorynebacterium diphtheriaebiovar intermedius with a predominant PFGE type. Most of them were toxigenic and some showed a decrease in penicillin G susceptibility. In conclusion, diphtheria remains endemic in Brazil. Health professionals need to be aware of the possibility of atypical cases ofC. diphtheriaeinfection, including pharyngitis without pseudomembrane formation.

Subinhibitory concentrations (subMICs) of antibiotics may alter bacterial surface properties and change microbial physiology. This study aimed to investigate the effect of a subMIC (⅛ MIC) of penicillin (PEN) and erythromycin (ERY) on bacterial morphology, haemagglutinating activity, cell-surface hydrophobicity (CSH) and biofilm formation on glass and polystyrene surfaces, as well as the distribution of cell-surface acidic anionic residues of Corynebacterium diphtheriae strains (HC01 tox − strain; CDC-E8392 and 241 tox + strains). All micro-organisms tested were susceptible to PEN and ERY. Growth in the presence of PEN induced bacterial filamentation, whereas subMIC of ERY caused cell-size reduction of strains 241 and CDC-E8392. Adherence to human erythrocytes was reduced after growth in the presence of ERY, while CSH was increased by a subMIC of both antibiotics in bacterial adherence to n-hexadecane assays. Conversely, antibiotic inhibition of biofilm formation was not observed. All strains enhanced biofilm formation on glass after treatment with ERY, while only strain 241 increased glass adherence after cultivation in the presence of PEN. Biofilm production on polystyrene surfaces was improved by ⅛ MIC of ERY. After growth in the presence of both antimicrobial agents, strains 241 and CDC-E8392 exhibited anionic surface charges with focal distribution. In conclusion, subMICs of PEN and ERY modified bacterial surface properties and enhanced not only biofilm formation but also cell-surface hydrophobicity. Antibiotic-induced biofilm formation may contribute to the inconsistent success of antimicrobial therapy for C. diphtheriae infections.

Corynebacterium pseudodiphtheriticum is a well-known human pathogen that mainly causes respiratory disease and is associated with high mortality in compromised hosts. Little is known about the virulence factors and pathogenesis of C. pseudodiphtheriticum. In this study, cultured human epithelial (HEp-2) cells were used to analyse the adherence pattern, internalisation and intracellular survival of the ATCC 10700 type strain and two additional clinical isolates. These microorganisms exhibited an aggregative adherence-like pattern to HEp-2 cells characterised by clumps of bacteria with a "stacked-brick" appearance. The differences in the ability of these microorganisms to invade and survive within HEp-2 cells and replicate in the extracellular environment up to 24 h post infection were evaluated. The fluorescent actin staining test demonstrated that actin polymerisation is involved in the internalisation of the C. pseudodiphtheriticum strains. The depolymerisation of microfilaments by cytochalasin E significantly reduced the internalisation of C. pseudodiphtheriticum by HEp-2 cells. Bacterial internalisation and cytoskeletal rearrangement seemed to be partially triggered by the activation of tyrosine kinase activity. Although C. pseudodiphtheriticum strains did not demonstrate an ability to replicate intracellularly, HEp-2 cells were unable to fully clear the pathogen within 24 h. These characteristics may explain how some C. pseudodiphtheriticum strains cause severe infection in human patients.

Corynebacterium diphtheriae is one of the most prominent human pathogens and the causative agent of the communicable disease diphtheria. The genomes of 12 strains isolated from patients with classical diphtheria, endocarditis, and pneumonia were completely sequenced and annotated. Including the genome of C. diphtheriae NCTC 13129, we herewith present a comprehensive comparative analysis of 13 strains and the first characterization of the pangenome of the species C. diphtheriae . Comparative genomics showed extensive synteny and revealed a core genome consisting of 1,632 conserved genes. The pangenome currently comprises 4,786 protein-coding regions and increases at an average of 65 unique genes per newly sequenced strain. Analysis of prophages carrying the diphtheria toxin gene tox revealed that the toxoid vaccine producer C. diphtheriae Park-Williams no. 8 has been lysogenized by two copies of the ω tox + phage, whereas C. diphtheriae 31A harbors a hitherto-unknown tox + corynephage. DNA binding sites of the tox -controlling regulator DtxR were detected by genome-wide motif searches. Comparative content analysis showed that the DtxR regulons exhibit marked differences due to gene gain, gene loss, partial gene deletion, and DtxR binding site depletion. Most predicted pathogenicity islands of C. diphtheriae revealed characteristics of horizontal gene transfer. The majority of these islands encode subunits of adhesive pili, which can play important roles in adhesion of C. diphtheriae to different host tissues. All sequenced isolates contain at least two pilus gene clusters. It appears that variation in the distributed genome is a common strategy of C. diphtheriae to establish differences in host-pathogen interactions.

O artigo revisa a literatura sobre a emergência de infecções humanas causadas por Corynebacterium ulcerans em diversos países, incluindo o Brasil. Foi realizada análise de artigos publicados entre 1926 e 2011 nas bases Medline/PubMed e SciELO, bem como artigos e informes do Ministério da Saúde. Apresenta-se um esquema de triagem, rápido, econômico e de fácil execução, capaz de permitir a realização do diagnóstico presuntivo de C. ulcerans e C. diphtheriae na maioria dos laboratórios brasileiros públicos e privados. A circulação de C. ulcerans em vários países, aliada aos recentes casos de isolamento do patógeno no Rio de Janeiro, é um alerta a clínicos, veterinários e microbiologistas sobre a ocorrência de difteria zoonótica e a circulação do C. ulcerans em regiões urbanas e rurais do território nacional e/ou da América Latina. The article is a literature review on the emergence of human infections caused by Corynebacterium ulcerans in many countries including Brazil. Articles in Medline/PubMed and SciELO databases published between 1926 and 2011 were reviewed, as well as articles and reports of the Brazilian Ministry of Health. It is presented a fast, cost-effective and easy to perform screening test for the presumptive diagnosis of C. ulcerans and C. diphtheriae infections in most Brazilian public and private laboratories. C. ulcerans spread in many countries and recent isolation of this pathogen in Rio de Janeiro, southeastern Brazil, is a warning to clinicians, veterinarians, and microbiologists on the occurrence of zoonotic diphtheria and C. ulcerans dissemination in urban and rural areas of Brazil and/or Latin America. El articulo revisa la literatura sobre la emergencia de infecciones humanas causadas por Corynebacterium ulcerans en diversos países, incluyendo Brasil. Se realizó análisis de artículos publicados entre 1926 y 2011 en las bases Medline/Pubmed y SciELO, así como artículos e informes del Ministerio Brasileño de la Salud. Se presenta un esquema de selección, rápido, económico y de fácil ejecución, capaz de permitir la realización del diagnóstico presuntivo de C. ulcerans y C. diphtheriae en la mayoría de los laboratorios brasileños públicos y privados. La circulación de C. ulcerans en varios países, aliada a los recientes casos de aislamiento del patógeno en Rio de Janeiro (Sureste de Brasil), es un alerta a clínicos, veterinarios y microbiólogos sobre la ocurrencia de difteria zoológica y la circulación de C. ulcerans en regiones urbanas y rurales del territorio nacional y/o de América Latina

We report a case of concurrent diphtheria and infectious mononucleosis in an 11-year-old Brazilian child. Two days after specific treatment for diphtheria was started the patient was discharged following clinical recovery. This case highlights the difficulties in the clinical diagnosis of diphtheria in partially immunized individuals, and for the management and control of diphtheria in developing countries.

The article is a literature review on the emergence of human infections caused by Corynebacterium ulcerans in many countries including Brazil. Articles in Medline/PubMed and SciELO databases published between 1926 and 2011 were reviewed, as well as articles and reports of the Brazilian Ministry of Health. It is presented a fast, cost-effective and easy to perform screening test for the presumptive diagnosis of C. ulcerans and C. diphtheriae infections in most Brazilian public and private laboratories. C. ulcerans spread in many countries and recent isolation of this pathogen in Rio de Janeiro, southeastern Brazil, is a warning to clinicians, veterinarians, and microbiologists on the occurrence of zoonotic diphtheria and C. ulcerans dissemination in urban and rural areas of Brazil and/or Latin America.

During the last decades, the majority of Brazilian Corynebacterium diphtheriae isolates were shown to be capable to metabolize sucrose, sometimes leading to erroneous identification as a non-diphtheric Corynebacterium species. The sequencing of the polymorphic region of the RNA polymerase beta subunit-encoding gene (rpoB) is an important taxonomic tool for identification of corynebacteria. The present study aimed to investigate the rpoB gene polymorphic features of sucrose-fermenting and non sucrose-fermenting strains. The results showed that sucrose-fermenting strains presented rpoB gene polymorphic regions with more than 98% similarity with the sequences deposited in the gene bank corresponding to non sucrose-fermenting strains. Data indicate that sucrose-fermenting isolates may act as a variant of C. diphtheriae biotype mitis. In addition we alert that sucrose-fermenting strains should not be discarded as contaminants mainly in countries where the possibility of isolation of this variant is higher.

The production of fibrinous exudates may play an important role in determining the outcome of bacterial infection. Although pseudomembrane formation is a characteristic feature of diphtheria, little is known about the fibrinogen (Fbn)-binding properties of Corynebacterium diphtheriae strains and the influence of the gene that codes for diphtheria toxin (tox gene) in this process. In this study we demonstrated the ability of C. diphtheriae strains to bind to Fbn and to convert Fbn to fibrin. Bacterial interaction with rabbit plasma was evaluated by both slide and tube tests. Interaction of microorganisms with human Fbn was evaluated by both enzyme linked immunosorbent assay (ELISA) and fluorescein isothiocyanate-conjugated (FITC) Fbn binding assays. Nontoxigenic and toxigenic strains formed bacterial aggregates in the presence of plasma in the slide tests. The ability to convert Fbn to a loose web of fibrin in the plasma solution in the tube tests appeared to be a common characteristic of the species, including strains that do not carry the tox gene. Fbn binding to C. diphtheriae strains occurred at varying intensities, as demonstrated by the FITC-Fbn and ELISA binding assays. Our data suggest that the capacity to bind to Fbn and to convert Fbn to fibrin may play a role in pseudomembrane formation and act as virulence determinants of both nontoxigenic and toxigenic strains.

A Tuberculose (TB) é uma doença infecto contagiosa que representa um dos principais problemas sociais, econômicos e de saúde publica no mundo. A tuberculose (TB) representa um dos principais problemas sociais, econômicos e de saúde pública no mundo. Em 2013 foram estimados que 9 milhões de pessoas desenvolveram tuberculose e 1,5 milhões de mortes associadas com epidemias de TB. Brasil está entre os 22 países que concentra 80% dos casos de TB no mundo.Esse trabalho teve como objetivo caracterizar o perfil epidemiológico dos pacientes com tuberculose pulmonar no período de 2008-2014 no estado do Maranhão. Trata-se de estudo descritivo-retrospectivo com abordagem quantitativa de casos novos de Tuberculose Pulmonar notificado ao Sistema de Informação de Agravos de Notificação (SINAN) ocorridos no Maranhão entre 2008 e 2014. Foram notificados 12.384 casos novos de tuberculose pulmonar. Nesse período, o Maranhão foi o décimo primeiro lugar no Brasil e o quarto da região Nordeste. Do mesmo modo, como acontece na população geral do Brasil e mundial, o sexo masculino apresenta maiores números de novos casos em comparação com o sexo feminino. A quantidade de pessoas acometidas por tuberculose esta na faixa etária de 20-39 anos (44,4%) Portanto, os pacientes com tuberculose pulmonar no maranhão no período de 2008 a 2014 apresentaram-se como sendo a maioria do sexo masculino com idade entre 20 e 39 anos, com ensino fundamental incompleto e de cor parda.Palavras-chave: Tuberculose pulmonar. Maranhão. Epidemiologia.