YOKENELLA REGENSBURGEI osteoarticular infection: a case report

okenella regensburgei belongs to the family Enterobacteriaceae and is an opportunistic agent rarely associated with infections in humans. We report a case of osteoarticular knee infection caused by Y. regensburgei in a patient under treatment for rheumatoid arthritis, using corticosteroids, with complication in primary total arthroplasty of the knee. Y. regensburgei was identified using the VITEK2 system. Antimicrobial susceptibility testing was performed using the disk-diffusion method, according to the guidelines from the Clinical and Laboratory Standards Institute. The patient presented favorable clinical evolution after the second debridement, with complete removal of the prosthesis and antibiotic therapy with sulfamethoxazole/trimethoprim. This is the first case of Y. regensburgei infection described d in Brazil.

Non-tuberculous mycobacterial lung disease: a brief review focusing on radiological findings

Abstract The incidence and prevalence of lung disease caused by non-tuberculous mycobacteria (NTM-LD) has increased worldwide and its diagnosis represents a complex challenge. This article aims to review the tomographic findings of NTM-LD in order to facilitate their definitive diagnosis. The search for publications on the subject was performed in PMC and Scielo using the keywords 'non-tuberculous mycobacteria', 'lung disease and computed tomography (CT)' and 'radiological findings'. The radiological findings described by 18 articles on mycobacteriosis were reviewed. In addition, CT images of patients diagnosed with NTM-LD were considered to represent radiological findings. Eighteen publications were used whose main findings were pulmonary cavitation (88.9%), bronchiectasis (77.8%), and pulmonary nodules (55.6%). Despite the overlaps in imaging-related analysis of myocobacterioses with other pulmonary infections, such as tuberculosis, the predominant involvement of the middle lobe and lingula should raise suspicion for NTM-LD.

Proteases associated with Mycobacterium tuberculosis infection

Tuberculosis is a contagious infectious disease caused by Mycobacterium tuberculosis, an obligate intracellular bacterium that relies on infection and host-to-host transmission to survive. In a co-evolutionary process, the pathogen developed virulence mechanisms to evade the host's immune system and endure a number of factors, such as cellular stress. One of the strategies used by pathogens to succeed in causing infection is the production of proteases, which are enzymes that cleave peptide bonds between the amino acids in a protein. Proteases are widely distributed in nature and have different roles considered important to the bacteria's biological cycle. M. tuberculosis has several protease coding genes in its genome, many of which with unknown functions, but several with recognized roles in the infection process. This review presents the literature researched from 2014 to 2018 that addressed the roles of the proteases involved in M. tuberculosis infection

Hyptis brevipes and Paullinia pinnata extracts and their fractions presenting activity against Mycobacterium abscessos SubSP. massiliense

Medicinal plants are of great interest for the discovery of new biomolecules with diversified effects. Over the last decade different outbreaks caused by Mycobacterium abscessos subsp. Massiliense have been reported, evidencing it as an important emerging pathogen in underdeveloped countries. This study investigated the antimycobacterial activity of six Brazilian medicinal plant extracts and their fractions. Hyptis brevipes, Tocoyena formosa, Randia armata, Paullinia pinnata, La foensia pacari And Anadenanthera colubrina were evaluated against M. a. massiliense. Total extracts from the medicinal plants H. brevipes, T. formosa, P. pinnata and L. pacari presented a minimal bactericidal concentration of 1 mg/mL. After fractioning, the ethanolic fractions from H. brevipes and P. pinnata presented bactericidal activity, and the ethyl acetate fraction from H. brevipes and T. formosa presented antimycobacterial action. The best bactericidal function of all plant fractions was the ethanolic, which contained rutin and rosmarinic acid that were shown to have microbicidal activity.

Isoniazid-resistant tuberculous spondylodiscitis associated with scrofuloderma

The extrapulmonary forms of tuberculosis are responsible for about 20% of cases. Scrofuloderma is the cutaneous manifestation secondary to infection in some subcutaneous foci. A 33-year-old patient was admitted to the Clinical Hospital with exudative skin lesions on the back and thorax, initiated 10 months previously, associated with daily fever, and constipation. Spine resonance showed a paravertebral pseudotumoral lesion with T4 and T9 invasion, including vertebral canal and sub-ligament extension. The lesions presented fistulas for paravertebral muscles, lung and skin. Polimerase chain reaction (PCR) proved positive for Mycobacterium tubeculosis in the thorax wound secretion, caracterizing tuberculous spondilodiscitis with scrofuloderma. Treatment was initiated with rifampicin, isoniazid, pyrazinamide and ethambutol with important clinical improvement after the first week. The febrile peaks came to an end and there was improvement in the pattern of the cutaneous lesions. The susceptibility test showed resistance to isoniazid

The mc2-CMX vaccine induces an enhanced immune response against Mycobacterium tuberculosis compared to Bacillus Calmette-Guérin but with similar lung inflammatory effects

Although the attenuated Mycobacterium bovis Bacillus Calmette-Guérin (BCG) vaccine has been used since 1921, tuberculosis (TB) control still proceeds at a slow pace. The main reason is the variable efficacy of BCG protection against TB among adults, which ranges from 0-80%. Subsequently, the mc2-CMX vaccine was developed with promising results. Nonetheless, this recombinant vaccine needs to be compared to the standard BCG vaccine. The objective of this study was to evaluate the immune response induced by mc2-CMX and compare it to the response generated by BCG. BALB/c mice were immunised with both vaccines and challenged withMycobacterium tuberculosis (Mtb). The immune and inflammatory responses were evaluated by ELISA, flow cytometry, and histopathology. Mice vaccinated with mc2-CMX and challenged with Mtb induced an increase in the IgG1 and IgG2 levels against CMX as well as recalled specific CD4+ T-cells that produced T-helper 1 cytokines in the lungs and spleen compared with BCG vaccinated and challenged mice. Both vaccines reduced the lung inflammatory pathology induced by the Mtb infection. The mc2-CMX vaccine induces a humoral and cellular response that is superior to BCG and is efficiently recalled after challenge with Mtb, although both vaccines induced similar inflammatory reductions.

Tuberculose latente em indivíduos de uma unidade prisional do centro oeste do Brasil / Latent tuberculosis among prison inmates in the brazilian mid-west

A tuberculose (TB) é a segunda principal causa de morte por doença infecciosa em todo o mundo e constitui um contínuo problema de saúde global. A ocorrência da TB ativa em indivíduos privados de liberdade (PL) é superior aos níveis médios relatados para a população geral. A descoberta tardia dos casos de TB, associada ao atraso no tratamento, agrava o problema da TB nas penitenciárias. Embora estas instituições sejam fechadas, a movimentação de prisioneiros entre diferentes ambientes da prisão, unidades prisionais ou instituições como tribunais transforma estes locais em verdadeiros reservatórios da TB. Este trabalho teve como objetivo fazer a triagem de voluntários PL para submetê-los à prova tuberculínica (PT). Os resultados desta primeira avaliação da PT em uma unidade prisional de Goiás evidenciaram positividade em 50,3% dos homens e 38,1% das mulheres. O acompanhamento dos dados destes indivíduos revelou que após um ano do recrutamento, 1,7% (n=9) dos voluntários apresentaram TB, dos quais quatro indivíduos pertenciam à mesma ala da unidade prisional masculina. A elevada taxa de infecção e adoecimento reforça a necessidade urgente de novas estratégias de identificação da TB latente, bem como de busca ativa nesta população

Methicillin-resistant Staphylococcus sp. colonizing health care workers of a cancer hospital

The aim of the study was to analyze epidemiological and microbiological aspects of oral colonization by methicillin-resistant Staphylococcus of health care workers in a cancer hospital. Interview and saliva sampling were performed with 149 health care workers. Antimicrobial resistance was determined by disk diffusion and minimum inhibitory concentration. Polymerase Chain Reaction, Internal Transcribed Spacer-Polymerase Chain Reaction and Pulsed Field Gel Electrophoresis were performed for genotypic characterization of methicillin-resistant Staphylococcus. Risk factors were determined by logistic regression. Methicillin-resistant Staphylococcus colonization prevalence was 19.5%, denture wearing (p = 0.03), habit of nail biting (p = 0.04) and preparation and administration of antimicrobial (p = 0.04) were risk factors identified. All methicillin-resistant Staphylococcus were S. epidermidis, 94.4% of them had mecA gene. Closely related and indistinguishable methicillin-resistant S. epidermidis were detected. These results highlight that HCWs which have contact with patient at high risk for developing infections were identified as colonized by MRSE in the oral cavity, reinforcing this cavity as a reservoir of these bacteria and the risk to themselves and patients safety, because these microorganisms may be spread by coughing and talking.

The use of Mycobacterium tuberculosis HspX and GlcB proteins to identify latent tuberculosis in rheumatoid arthritis patients

Rheumatoid arthritis (RA) is an autoimmune disease characterised by the destruction of articular cartilage and bone damage. The chronic treatment of RA patients causes a higher susceptibility to infectious diseases such as tuberculosis (TB); one-third of the world’s population is latently infected (LTBI) with Mycobacterium tuberculosis (Mtb). The tuberculin skin test is used to identify individuals LTBI, but many studies have shown that this test is not suitable for RA patients. The goal of this work was to test the specific cellular immune responses to the Mtb malate synthase (GlcB) and heat shock protein X (HspX) antigens of RA patients and to correlate those responses with LTBI status. The T-helper (Th)1, Th17 and Treg-specific immune responses to the GlcB and HspX Mtb antigens were analysed in RA patients candidates for tumour necrosis factor-α blocker treatment. Our results demonstrated that LTBI RA patients had Th1-specific immune responses to GlcB and HspX. Patients were followed up over two years and 14.3% developed active TB. After the development of active TB, RA patients had increased numbers of Th17 and Treg cells, similar to TB patients. These results demonstrate that a GlcB and HspX antigen assay can be used as a diagnostic test to identify LTBI RA patients.

Methicillin-resistant Staphylococcus aureus nasal carriage in neonates and children attending a pediatric outpatient clinics in Brazil

Background: In Latin America, few studies have been carried out on methicillin-resistant Staphylococcus aureus carriage in the pediatric population. We conducted a survey of nasal S. aureus carriage in neonates and in children attending the pediatric outpatient clinics in a large Brazilian city with high antimicrobial consumption. Methods: Pernasal swabs of neonates were collected upon admission and at discharge in four neonatal intensive care units and of children less than five years of age during outpatient visits. Methicillin-resistant S. aureus isolates were characterized for antibiotic susceptibility, mec gene presence, pulsed-field gel electrophoresis, spa type, SCCmec-type, multilocus sequence type, and presence of Panton-Valentine leukocidin genes. Results: S. aureus was carried by 9.1% and 20.1% of the 701 neonates and of 2034 children attending the outpatient clinics, respectively; methicillin-resistant S. aureus carriage was detected in 0.6% and 0.2%, of the these populations, respectively. Healthcare-associated methicillin-resistant S. aureus strains found in neonates from neonatal intensive care units and outpatients were genetically related to the Brazilian (SCCmec-III, ST239) and to the Pediatric (SCCmec-IV, ST5) clones. Community-associated methicillin-resistant S. aureus was only detected in outpatients. None of the methicillin-resistant S. aureus strains contained the Panton-Valentine leukocidin gene. Methicillin-resistant S. aureus strains related to the Brazilian clone showed multidrug resistance pattern. Conclusions: Despite the high antibiotic pressure in our area, and the cross transmission of the healthcare-associated methicillin-resistant S. aureus clones between neonatal intensive care units and outpatients, the prevalence of methicillin-resistant S. aureus carriage is still low in our setting. .

Cellular immune response of Curraleiro Pé-duro and Nellore calves following Mycobacterium bovis-BCG vaccination / Resposta imune celular de bezerros Curraleiro Pé-duro e Nelore após vacinação com Mycobacterium bovis-BCG

The present study aimed to assess the CD4, CD8 and Gamma delta T cells blood levels for Curraleiro Pé-duro, as well as the specific IFN-γ response after BCG vaccination using flow cytometry. The specific immune response against BCG was also evaluated by tuberculin skin test, performed before and 45 days after the vaccination. For comparison purposes, the same parameters were investigated on Nellore calves, an exotic bovine with resistance previously demonstrated. Naturally, Curraleiro Pé-duro animals had greater levels of CD4, CD8 and Gamma delta lymphocytes (p<0.05). In response to vaccine, Curraleiro Pé-duro showed greater ability to respond specifically to BCG, generating resistance profile (Th1), evidenced by greater number of antigen specific CD4+ cells producing IFN-γ (p<0.05) and also higher tuberculin skin test reaction (p<0.05). Additionally, vaccinated Curraleiro Pé-duro calves had higher CD4 cells numbers than both Nellore control (p<0.05) and vaccinated groups (p<0.05). Curraleiro Pé-duro calves' higher basal lymphocytes blood level and stronger response in both IFN-γ and tuberculin skin test parameters probably play a positive role on protection/resistance to Mycobacterium bovis.

O presente estudo teve como objetivo avaliar níveis sanguíneos de células CD4, CD8 e células T gama-delta no sangue periférico de bezerros Curraleiro Pé-Duro, bem como a produção específica de IFN-γ por essas células em resposta à vacinação com BCG, através de citometria de fluxo. A resposta imune específica contra BCG também foi avaliada por teste tuberculínico, realizado antes e 45 dias após a vacinação. Para fins de comparação, os mesmos parâmetros foram investigados em bezerros da raça Nelore, uma raça bovina exótica com resistência demonstrado anteriormente. Naturalmente, animais da raça Curraleiro Pé-Duro apresentaram maiores níveis de CD4, CD8 e linfócitos gama-delta. Em resposta a vacina, Curraleiro Pé-duro mostrou maior capacidade de responder especificamente ao BCG, gerando perfil de resistência (Th1), evidenciado pelo maior número de células CD4+ específicas produtoras de IFN-γ e maior reação cutânea a por tuberculina. Os maiores níveis basais de linfócitos, maior produção de IFN-γ e reação cutânea à prova tuberculínica provavelmente desempenham um papel positivo na proteção/resistência ao Mycobacterium tuberculosis.

Clinical data and molecular analysis of Mycobacterium tuberculosi isolates from drug-resistant tuberculosis patients in Goiás, Brazil

Drug resistance is one of the major concerns regarding tuberculosis (TB) infection worldwide because it hampers control of the disease. Understanding the underlying mechanisms responsible for drug resistance development is of the highest importance. To investigate clinical data from drug-resistant TB patients at the Tropical Diseases Hospital, Goiás (GO), Brazil and to evaluate the molecular basis of rifampin (R) and isoniazid (H) resistance in Mycobacterium tuberculosis. Drug susceptibility testing was performed on 124 isolates from 100 patients and 24 isolates displayed resistance to R and/or H. Molecular analysis of drug resistance was performed by partial sequencing of the rpoB and katGgenes and analysis of the inhA promoter region. Similarity analysis of isolates was performed by 15 loci mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) typing. The molecular basis of drug resistance among the 24 isolates from 16 patients was confirmed in 18 isolates. Different susceptibility profiles among the isolates from the same individual were observed in five patients; using MIRU-VNTR, we have shown that those isolates were not genetically identical, with differences in one to three loci within the 15 analysed loci. Drug-resistant TB in GO is caused by M. tuberculosis strains with mutations in previously described sites of known genes and some patients harbour a mixed phenotype infection as a consequence of a single infective event; however, further and broader investigations are needed to support our findings.

New vectors derives from pUC 18 for clonig and thermal-induced expression in Escherichia coli

We report the construction of two vectors for Escherichia coli: pUC72, for molecular cloning, and pPLT7, for thermal-induced expression. The main feature of pUC72 is a novel polylinker region that includes restriction sites for Nde I and Nco I which provide an ATG codon for proper translation initiation of expressed genes. Vector pPLT7 is ideal for thermo-inducible expression in host cells that carry the cI857 repressor gene. The use of pPLT7 was validated by the successful expression of the genes encoding carp and porcine growth hormones. These vectors provide novel cloning possibilities in addition to simple, non-expensive, high level expression of recombinant proteins in E. coli.

Health care workers humoral immune response against GLcB, MPT51 and HSPX from Mycobacterium tuberculosis

Tuberculosis (TB) is one of the oldest human infectious diseases and one third of the world's population is latently infected. Brazil is an endemic area for TB. One of the most important challenges in TB control is the identification of latently infected individuals. Health Care Workers (HCW) are at high risk of being infected with Mycobacterium tuberculosis and even to become TB latently infected. The aim of this study was to increase knowledge about humoral immune response in TB latently infected individuals. HCW were classified according to their tuberculin skin test (TST), as positive or negative. The antibody response to GLcB, MPT51 and HSPX from Mycobacterium tuberculosis was evaluated. TST negative HCW constituted the majority of those who showed a humoral immune response. Antibody levels varied according to antigen characteristics, TST and BCG status. We suggest that possibly the presence of those antibodies could have a function in the protective immune response against Mycobacterium tuberculosis.

Limitations of the BCG vaccine and new prophylaxis strategies against human tuberculosis / Limitações da vacina BCG e novas estratégias profiláticas contra tuberculose humana

A atual vacina contra a tuberculose, o BCG (Bacilo Calmette Guérin), uma vacina atenuada, derivada do Mycobacterium bovis, apesar de proteger as crianças contra a enfermidade, falha na proteção contra a tuberculose pulmonar ativa em adultos, principalmente em países onde a doença é endêmica. Uma nova vacina para tuberculose deve proteger várias categorias de indivíduos, como crianças, adultos, idosos e imunocomprometidos. Sendo assim, uma característica importante a se considerar é a seguridade vacinal para todas as classes de imunizados. Esta revisão propõe apresentar as novas estratégias de vacinação, tais como subunidades vacinais, vacinas de DNA, vacinas com micro-organismos e vetores vivos e discutir as aplicações dessas novas estratégias no controle e erradicação da tuberculose.

Detecção do gene mecA em estafilococos coagulase negativa resistentes à oxacilina isolados da saliva de profissionais da enfermagem / Detection of mecA gene in oxacillin-resistant coagulase-negative staphylococci isolated from the saliva of nursing professionals

Estafilococos coagulase negativa estão frequentemente associados às infecções nosocomiais e os profissionais da saúde podem ser reservatório e dissemina-los no hospital e comunidade. O objetivo desse estudo foi identificar espécies de estafilococos coagulase negativa isolados da saliva de profissionais da enfermagem, determinar o perfil de resistência e detectar o gene mecA. Foram selecionados 100 estafilococos coagulase negativa, sendo 41 identificados como Staphylococcus epidermidis, 25 Staphylococcus saprophyticus, 18 Staphylococcus haemolyticus, 8 Staphylococcus cohnii, 4 Staphylococcus lugdunenses, 3 Staphylococcus capitis, e 1 Staphylococcus Simulans. Desses, 32 por cento apresentaram resistência à oxacilina, 84,4 por cento à mupirocina, 32 por cento à cefoxitina, e todos sensíveis a vancomicina. Dos estafilococos coagulase negativa resistentes à oxacilina, 93,7 por cento desenvolveram-se no agar oxacilina (6µg/ml) e o gene mecA foi detectado em 75 por cento. Os resultados sinalizam que maiores investimentos devem ser direcionados a identificação das espécies de estafilococos coagulase negativa nas instituições de saúde e na comunidade.

Coagulase-negative staphylococci are frequently associated with nosocomial infections, and healthcare professionals can be reservoirs and spread them in hospitals and in the community. The aim of this study was to identify species of coagulase-negative staphylococci isolated from the saliva of nursing professionals, determine the resistance profile and detect the mecA gene. One hundred coagulase-negative staphylococci were selected: 41 were identified as Staphylococcus epidermidis, 25 as Staphylococcus saprophyticus, 18 as Staphylococcus haemolyticus, eight as Staphylococcus cohnii, four as Staphylococcus lugdunenses, three as Staphylococcus capitis and one as Staphylococcus simulans. Of these, 32 percent presented oxacillin resistance, 84.4 percent mupirocin resistance and 32 percent cefoxitin resistance, and all were vancomycin sensitive. Among the oxacillin-resistant coagulase-negative staphylococci, 93.7 percent developed in oxacillin agar (6µg/ml) and the mecA gene was detected in 75 percent. The results indicate that higher investments should be directed towards identifying coagulase-negative staphylococcus species in healthcare institutions and in the community.

Extensively drug-resistant tuberculosis: a case report and literature review

Extensively drug-resistant tuberculosis (XDR-TB) is an emerging health problem that threatens tuberculosis (TB) control worldwide, since suitable treatment for this disease has not yet been found. We report a case of secondary pulmonary XDR-TB in a 54-year-old, HIV-negative male from Goiânia, Brazil. The patient had long-standing pulmonary tuberculosis (nine years) with extensive bilateral lung damage and had been treated with multiple antituberculosis drugs (self-administered) before XDR-TB diagnosis. The strain of Mycobacterium tuberculosis was resistant to R- rifampicin, H-isoniazid, E-ethambutol, Eto-ethionamide, Ofx-ofloxacin, and Am-amikacin. This patient died with multiple organ failure due to sepsis secondary to bacterial pneumonia.

Multidrug-resistant tuberculosis: case reports study in a central state of Brazil

Multidrug-resistant tuberculosis (MDR-TB) is an emerging and worrisome health problem that threatens tuberculosis (TB) control worldwide. The clinical management of MDR-TB is a complex issue associated with the use of multiple drugs for a long period, usually accompanied by side effects and high costs. The objective of this work was to relate cases of MDR-TB occurring in Goiás, a central state of Brazil. We related five cases of MDR-TB, three women and two men. All were pulmonary cases. Three were in their second treatment and two in their first treatment. Surgical pulmonary resection was performed in one case. One death occurred. Lack of adherence, gastric intolerance to anti-TB drugs and poor clinical management were the main aspects related to the emergent resistance. A revision of the main clinical aspects of this disease was performed.

Métodos aplicados à epidemiologia molecular do Mycobacterium tuberculosis / Methods employed in the molecular epidemiology of Mycobacterium tuberculosis

Estima-se que um terço da população mundial esteja infectada com Mycobacterium tuberculosis, resultando em dois milhões de mortes anualmente. No mundo, são registrados mais de oito milhões de novos casos de tuberculose por ano e o Brasil é um dos principais países nos quais ocorrem tais registros. Os fatores determinantes para o controle da tuberculose são: a detecção rápida, a terapia adequada e os meios de se evitar futuras transmissões. A caracterização de linhagens por tipagem molecular é um instrumento útil nessas investigações epidemiológicas. Existem vários métodos de tipagem molecular para a caracterização destas linhagens. O IS6110-RFLP é a técnica padronizada mundialmente. Algumas técnicas, como spoligotyping, VNTRs-MIRUs, entre outras, têm representado alternativas para análises de isolados de M.tuberculosis geneticamente relacionados, especialmente em casos em que a análise de IS6110-RFLP não é aplicável. Nesta revisão, foram consultadas publicações relevantes sobre o assunto, em periódicos especializados, a fim de se obter maior compreensão sobre as técnicas de genotipagem do Mycobacterium sp e suas respectivas vantagens e limitações.