Occurrence and antimicrobial resistance profile of bacteria isolated from bronchoalveolar lavage of patients admitted to hospitals in Fortaleza in the period from January 1996 to December 2001

Justificativa â A pneumonia hospitalar (PH) à causa de morbidade e mortalidade elevadas em pacientes hospitalizados. A terapia antimicrobiana empÃrica adequada e precoce pode salvar a vida de mais da metade dos pacientes com PH e deve ser baseada em padrÃes locais de sensibilidade a antimicrobianos. Praticamente todo tratamento de PH Ã, inicialmente, empÃrico. O objetivo deste trabalho à contribuir com o conhecimento do padrÃo regional de resistÃncia de microrganismos nesse contexto. Metodologia â Foram estudados 588 resultados de culturas de lavado bronco-alveolar (LBA) de pacientes internados em Fortaleza, processados na rotina de um laboratÃrio privado, no perÃodo de janeiro de 1996 a dezembro de 2001. Como resultado de pesquisa aos prontuÃrios mÃdicos desses pacientes, dois grupos foram criados: Grupo 1, com n=147, composto por pacientes com pneumonia hospitalar (PH) notificada segundo os critÃrios do Center for Disease Control and Prevention (CDC); Grupo 2, com n=382, pacientes com pneumonia nÃo-hospitalar (PNH). Utilizou-se a tÃcnica quantitativa de cultivo descrita nos trabalhos de Kahn e Jones (1987), Salata et al. (1987) e Carvalho (1997). IdentificaÃÃo e antibiogramas foram realizados no equipamento VITEK BioMerrieux, exceto para Streptococcus pneumoniae e Stenotrophomonas maltophilia cujos antibiogramas foram realizados pelo mÃtodo Kirby-Bauer e E-test respectivamente. Resultados â No Grupo 1, os cinco microrganismos mais freqÃentes foram Pseudomonas aeruginosa [56 casos (38,1%)], Staphylococcus aureus [24 casos (16,3%)], Klebsiella pneumoniae [12 casos (8,2%)], Acinetobacter spp [12 casos (8,2%)] e Serratia marcescens [10 casos (6,8%)]. No Grupo 2, encontraram-se, mais freqÃentemente, Pseudomonas aeruginosa [113 casos (29,6%)], Staphylococcus aureus [89 casos (23,3%)], Klebsiella pneumoniae [32 casos (8,4%)] e Acinetobacter spp [31 casos (8,1%)]. NÃo foi observada diferenÃa significativa entre os dois grupos para a etiologia. O mesmo ocorreu com o perfil de resistÃncia dos organismos, exceto para o Grupo 1 com S. aureus/oxacilina (p=0,027) e P. aeruginosa/piperacilina/tazobactam (p=0,003). No perfil de resistÃncia do conjunto total de amostras (n=751), destaca-se a de P. aeruginosa ao imipenem, de 40,8%; de Acinetobacter spp ao imipenem, de 10,0%; de Acinetobacter spp a Ampicilina/Sulbactam, de 44,3%; e de S. aureus a oxacilina, de 67,3%. O intervalo de tempo entre a data de internaÃÃo e a realizaÃÃo da cultura foi maior do que 7 dias em 60,18% dos casos. ConclusÃo - no trato respiratÃrio, o problema da multiresistÃncia bacteriana à evidente e preocupante com alta prevalÃncia de bacilos gram-negativos multiresistentes, marcadamente P. aeruginosa e Acinetobacter spp., assim como elevada resistÃncia a oxacilina nas amostras de Staphylococcus aureus. O Grupo 2 nÃo possui caracterÃsticas de pneumonia comunitÃria (PC), provavelmente, porque o tempo entre a internaÃÃo e a realizaÃÃo da cultura foi longo o suficiente para permitir a colonizaÃÃo do trato respiratÃrio superior pela microflora do ambiente hospitalar. à possÃvel que o Grupo 2 seja constituÃdo por pacientes com pneumonia comunitÃria severa refratÃria à antibioticoterapia que necessita internaÃÃo, ou que tiveram vÃrias internaÃÃes anteriores, com conseqÃente colonizaÃÃo por microflora hospitalar. InvestigaÃÃes subseqÃentes devem conferir atenÃÃo especial a esse aspecto. Pode ser Ãtil, neste contexto, o emprego de tÃcnicas de Biologia Molecular para melhor caracterizaÃÃo dos microrganismos isoladosHospital acquired pneumonia (HAP) is associated with high morbidity and mortality in hospitalized patients. Early, appropriate, and adequate empiric antibiotic therapy can save lives of more than half of all HAP patients and must be based on local data. This study will provide local patterns of antibiotic resistance. Practically all primary therapy of HAP is empiric and information from surveillance program of a given hospital is very important. We studied 588 Bronchoalveolar lavage cultures results from hospitalized patients performed in a private lab during a period of 6 years from 1996 to 2001. As a result of searching patientâs records, two groups were assigned: Group 1, n=147, patients with HAP notified by Nosocomial Infection Commission according to Center for Disease Control and Prevention-CDC; Group 2, n=382, patients with No-Nosocomial Pneumonia. Bacteriologic cultures were done quantitatively with a threshold of >= 105 according to Kahn and Jones (1987), Salataet al. (1987) and Carvalho (1997). Identification and susceptibility tests were performed on VITEK BioMerrieux except for Streptococcus pneumoniae and Stenotrophomonas maltophilia. In patients from Group 1, the five most frequent agents were: P. aeruginosa 56 cases (38,1%), S.aureus 24 (16,3%), K. pneumonia 12 (8,2%), Acinetobacter spp 12 (8,2%) and S. marcescens 10 (6,8%). Group 2 shows: P. aeruginosa 113 (29,6%), Staphylococcus aureus 89 (23,3%), Klebsiella pneumoniae 32 (8,4%), Acinetobacter spp 31 (8,1%) and Candida spp 20 (5,2%). There was no significant difference between resistance profile of isolates when distributed in two groups except S. aureus/Oxacilina (p=0,027), P.aeruginosa/Piperacilina/Tazobactam (p=0,003). The resistance profile in total (n=751) was: P. aeruginosa/Imipenem 40,8%, Acinetobacter spp/Imipenem 10,0%, Acinetobacter spp/AmpicilinaSulbactam 44,3% e S. aureus/Oxacilina 67,3%. The time between admission date and culture request was more than 7days in 60,18% in both groups. Conclusion: a) drug-resistance in lower respiratory tract infection(LRTI) is a serious concern mainly with high prevalence of multi-R gram-negative like P. aeruginosa and Acinetobacter with high resistance for Imipenem and other β- IactÃmic and S. aureus with high resistance to Oxacilina. There was no significant difference between the two groups. Group 2 did not show characteristics of Community-Acquired Pneumoniae (CAP) maybe because of large intervals of time between admission and request of culture, or this kind of patient had either severe CAP with no response to prior multiple antimicrobial therapy or previous hospitalizations or even had recent hospitalization and consequent colonization. Forwards molecular studies should be performed on isolates to provide better characterization of lower respiratory tract pathogens

Simvastatin inhibits planktonic cells and biofilms of Candida and Cryptococcus species

The antifungal activity of some statins against different fungal species has been reported. Thus, at the first moment, the in vitro antifungal activity of simvastatin, atorvastatin and pravastatin was tested against Candida spp. and Cryptococcus spp. Then, in a second approach, considering that the best results were obtained for simvastatin, this drug was evaluated in combination with antifungal drugs against planktonic growth and tested against biofilms of Candida spp. and Cryptococcus spp. Drug susceptibility testing was performed using the microdilution broth method, as described by the Clinical and Laboratory Standards Institute. The interaction between simvastatin and antifungals against planktonic cells was analyzed by calculating the fractional inhibitory concentration index. Regarding biofilm susceptibility, simvastatin was tested against growing biofilm and mature biofilm of one strain of each tested yeast species. Simvastatin showed inhibitory effect against Candida spp. and Cryptococcus spp. with minimum inhibitory concentration values ranging from 15.6 to 1000 mg L−1 and from 62.5 to 1000 mg L−1, respectively. The combination of simvastatin with itraconazole and fluconazole showed synergism against Candida spp. and Cryptococcus spp., while the combination of simvastatin with amphotericin B was synergistic only against Cryptococcus spp. Concerning the biofilm assays, simvastatin was able to inhibit both growing biofilm and mature biofilm of Candida spp. and Cryptococcus spp. The present study showed that simvastatin inhibits planktonic cells and biofilms of Candida and Cryptococcus species. Keywords: Candida, Cryptococcus, Simvastatin, Antifungal activity, Biofil

Easy Storage Strategies for Sporothrix spp. Strains

The present study evaluated the maintenance of Sporothrix spp. (6 Sporothrix brasiliensis; 6 S. schenckii; 5 S. mexicana, and 3 S. globosa) in saline at 4 degrees C, and in 10% glycerol plus either 10% lactose or 10% sucrose, at -20 degrees C and -80 degrees C. Viability was assessed after 3, 6, and 9 months of storage, through the recovery of strains on potato dextrose agar and analysis of macro- and micromorphological features. Conidium quantification was performed before and after storage, at 3, 6 and 9 months. 100% viability was observed, regardless of storage conditions or time period. Storage at 4 degrees C and at -20 degrees C did not alter the number of conidia, but lower conidium counts were observed at -80 degrees C. This study shows that the combination of glycerol with lactose or sucrose is effective to maintain Sporothrix spp. at freezing temperatures.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Univ Fed Ceara, Grad Program Med Microbiol, Specialized Med Mycol Ctr, Fortaleza, Ceara, BrazilUniv Fed Ceara, Sch Med, Christus Coll, UNICHRISTUS, Fortaleza, Ceara, BrazilUniv Fed Ceara, Dept Stat & Appl Math, Fortaleza, Ceara, BrazilUniv Fed Ceara, Grad Program Med Sci, Fortaleza, Ceara, BrazilUniv Estadual Ceara, Grad Program Vet Sci, Fortaleza, Ceara, BrazilUniversidade Federal de São Paulo, Dept Microbiol Immunol & Parasitol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Microbiol Immunol & Parasitol, São Paulo, BrazilCNPq: PROTAX 562296/2010-7CNPq: 304779/2011-3CNPq: 504189/2012-3Web of Scienc

Macrobrachium amazonicum: an alternative for microbiological monitoring of aquatic environments in Brazil

This study aimed to evaluate the role of the Amazon River prawn, Macrobrachium amazonicum, as carrier of Candida spp., by analyzing the correlation between Candida spp. from these prawns and their environment (surface water and sediment), through M13-PCR fingerprinting and RAPD-PCR. For this purpose, 27 strains of Candida spp. were evaluated. These strains were recovered from the gastrointestinal tract of adult M. amazonicum (7/27) from Catú Lake, Ceará State, Brazil and from the aquatic environment (surface water and sediment) of this lake (20/27). Molecular comparison between the strains from prawns and the aquatic environment was conducted by M13-PCR fingerprinting and RAPD-PCR, utilizing the primers M13 and OPQ16, respectively. The molecular analysis revealed similarities between the band patterns of eight Candida isolates with the primer M13 and 11 isolates with the primer OPQ16, indicating that the same strains are present in the digestive tract of M. amazonicum and in the aquatic environment where these prawns inhabit. Therefore, these prawns can be used as sentinels for environmental monitoring through the recovery of Candida spp. from the aquatic environment in their gastrointestinal trac

Terpinen-4-ol, tyrosol, and beta-lapachone as potential antifungals against dimorphic fungi

This study aimed to evaluate the in vitro antifungal activity of terpinen-4-ol, tyrosol, and beta-lapachone against strains of Coccidioides posadasii in filamentous phase (n=22) and Histoplasma capsulatum in both filamentous (n=40) and yeast phases (n=13), using the broth dilution methods as described by the Clinical and Laboratory Standards Institute, to determine the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of these compounds. The mechanisms of action of these compounds were also investigated by analyzing their effect on cell membrane permeability and ergosterol synthesis. The MIC and MFCf these compounds against C. posadasii, mycelial H. capsulatum, and yeast-like H. capsulatum, were in the following ranges: 350-5720 mu g/mL, 20-2860 mu g/mL, and 40-1420 mu g/mL, respectively for terpinen-4-ol250-4000 mu g/mL, 30-2000 mu g/mL, and 10-1000 mu g/mL, respectively, for tyrosoland 0.48-7.8 mu g/mL, 0.25-16 mu g/mL, and 0.125-4 mu g/mL, respectively for p-lapachone. These compounds showed a decrease in MIC when the samples were subjected to osmotic stress, suggesting that the compounds acted on the fungal membrane. All the compounds were able to reduce the ergosterol content of the fungal strains. Finally, tyrosol was able to cause a leakage of intracellular molecules. (C) 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Univ Fed Ceara, Postgrad Program Med Microbiol, Specialized Med Mycol Ctr, Fortaleza, Ceara, BrazilUniv Fed Ceara, Postgrad Program Med Sci, Fortaleza, Ceara, BrazilUniv Fed Sao Paulo, Dept Microbiol Immunol & Parasitol, Cellular Biol Div, Sao Paulo, SP, BrazilUniv Fed Ceara, Dept Stat & Appl Math, Fortaleza, Ceara, BrazilChristus Coll Unichristus, Sch Med, Fortaleza, Ceara, BrazilUniv Estadual Ceara, Postgrad Program Vet Sci, Fortaleza, Ceara, BrazilUniversidade Federal de São Paulo (UNIFESP), Department of Microbiology, Immunology and Parasitology, Cellular Biology Division, São Paulo, SP, BrazilCNPq: 303396/2014-8CNPq: 552161/2011-0CAPES: AE1-0052-000630100/11Web of Scienc