Yeast isolation and identification in water used in a Brazilian hemodialysis unit by classic microbiological techniques and Raman spectroscopy.

The use of poorly treated water in hemodialysis centers may lead to fungal contamination, which poses a serious threat to immunologically debilitated hemodialysis patients. This study aimed to isolate and identify yeast species in the water of a Brazilian hemodialysis center by using classic microbiological techniques and Raman spectroscopy. For 12 months, a total of 288 water samples were collected from different points of the hemodialysis treatment distribution center. One hundred and forty-six yeast species were isolated and identified in the samples that tested positive for the presence of yeasts such as Candida parapsilosis (100 isolates, or 68.50%), C. guilliermondii (17 isolates, or 11.65%), Rhodotorula mucilaginosa (23 isolates, or 15.75%), R. glutinis (three isolates, or 2.05%), and Trichosporon inkin (three isolates, or 2.05%). Yeast susceptibility to the antifungal fluconazole was also assayed. Only two C. guilliermondii isolates were resistant to fluconazole: the minimal inhibitory concentrations were higher than 64 µg/mL. The different yeast species present in the water of a Brazilian hemodialysis center call for more effective water disinfection procedures in this unit. Raman spectroscopy is an excellent tool to identify yeast species and is potentially applicable in routine water monitoring in hemodialysis units.

Isolation and identification of environmental mycobacteria in the waters of a hemodialysis center.

The use of poorly treated water during hemodialysis may lead to contamination with nontuberculous mycobacteria (NTM). This study aimed to isolate and identify NTM species in the water of a Brazilian hemodialysis center. We collected 210 samples of water from the hydric system of the unit (post-osmosis system, hemodialysis rooms, reuse system, and hemodialysis equipment) and from the municipal supply network; we isolated the NTM by a classic microbiological technique and identified them by the PCR restriction enzyme pattern of the hsp65 gene (PRA). Fifty-one (24.3 %) of the collected samples tested positive for NTM; both the municipal supply network (2 samples, 3.2 %) and the hydric system of the hemodialysis center (49 samples, 96.1 %) contained NTM. We isolated and identified potentially pathogenic bacteria such as Mycobacterium lentiflavum (59.0 %) and M. kansasii (5.0 %), as well as rarely pathogenic bacteria like M. gordonae (24.0 %), M. gastri (8.0 %), and M. szulgai (4.0 %). The ability of NTM to cause diseases is well documented in the literature. Therefore, the identification of NTM in the water of a Brazilian hemodialysis center calls for more effective water disinfection procedures in this unit.

Anticandidal efficacy of cinnamon oil against planktonic and biofilm cultures of Candida parapsilosis and Candida orthopsilosis.

Candida parapsilosis is yeast capable of forming biofilms on medical devices. Novel approaches for the prevention and eradication of the biofilms are desired. This study investigated the anticandidal activity of sixteen essential oils on planktonic and biofilm cultures of C. parapsilosis complex. We used molecular tools, enumeration of colony-forming units, the colourimetric MTT assay, scanning electron microscopy (SEM) and a chequerboard assay coupled with software analyses to evaluate the growth kinetics, architecture, inhibition and reduction in biofilms formed from environmental isolates of the Candida parapsilosis complex; further, we also evaluated whether essential oils would interact synergistically with amphotericin B to increase their anticandidal activities. Of the environmental C. parapsilosis isolates examined, C. parapsilosis and C. orthopsilosis were identified. Biofilm growth on polystyrene substrates peaked within 48 h, after which growth remained relatively stable up to 72 h, when it began to decline. Details of the architectural analysis assessed by SEM showed that C. parapsilosis complex formed less complex biofilms compared with C. albicans biofilms. The most active essential oil was cinnamon oil (CO), which showed anticandidal activity against C. orthopsilosis and C. parapsilosis in both suspension (minimum inhibitory concentration-MIC-250 and 500 µg/ml) and biofilm (minimum biofilm reduction concentration-MBRC-1,000 and 2,000 µg/ml) cultures. CO also inhibited biofilm formation (MBIC) at concentrations above 250 µg/ml for both species tested. However, synergism with amphotericin B was not observed. Thus, CO is a natural anticandidal agent that can be effectively utilised for the control of the yeasts tested.

Determination of the antibacterial activity of crude extracts and compounds isolated from Hortia oreadica (Rutaceae) against oral pathogens / Determinação da atividade antibacteriana de extratos brutos e substâncias isoladas de Hortia oreadica (Rutaceae) frente à patógenos bucais

Extracts from Hortia oreadica afforded four dihydrocinnamic acid derivatives, isolated from the n-hexane extract, as well as limonoid guyanin and the furoquinoline alkaloid dictamnine, both isolated from the dichloromethane extract. The extracts and the isolated compounds were tested against some oral pathogens, so as to investigate their antibacterial activity. The results showed that the n-hexane extract and the compound dictamnine are the most active against the selected microorganisms.

Extratos brutos de Hortia oreadica, forneceram quatro derivados do ácido diidrocinâmico, que foram isolados do extrato n-hexânico, bem como as substâncias guianina e dictamina, isoladas do extrato em diclorometano. Os extratos brutos e as substâncias isoladas foram avaliados frente a alguns patógenos bucais com o objetivo de investigar a atividade antibacteriana. Os resultados demonstraram que o extrato bruto n-hexânico e a substância dictamina foram os mais ativos frente ao conjunto de microrganismos avaliados.

Microbiological contamination of a hemodialysis center water distribution system.

The microbiological monitoring of the water used for hemodialysis is extremely important, especially because of the debilitated immune system of patients suffering from chronic renal insufficiency. To investigate the occurrence and species diversity of bacteria in waters, water samples were collected monthly from a hemodialysis center in upstate São Paulo and tap water samples at the terminal sites of the distribution system was sampled repeatedly (22 times) at each of five points in the distribution system; a further 36 samples were taken from cannulae in 19 hemodialysis machines that were ready for the next patient, four samples from the reuse system and 13 from the water storage system. To identify bacteria, samples were filtered through 0.22 microm-pore membranes; for mycobacteria, 0.45 microm pores were used. Conventional microbiological and molecular methods were used in the analysis. Bacteria were isolated from the distribution system (128 isolates), kidney machine water (43) and reuse system (3). Among these isolates, 32 were Gram-positive rods, 120 Gram-negative rods, 20 Gram-positive cocci and 11 mycobacteria. We propose the continual monitoring of the water supplies in hemodialysis centers and the adoption of effective prophylactic measures that minimize the exposure of these immunodeficient patients to contaminated sources of water.

Microbiological contamination of a hemodialysis center water distribution system / Contaminação microbiológica no sistema de distribuição de água de um centro de hemodiálise

The microbiological monitoring of the water used for hemodialysis is extremely important, especially because of the debilitated immune system of patients suffering from chronic renal insufficiency. To investigate the occurrence and species diversity of bacteria in waters, water samples were collected monthly from a hemodialysis center in upstate São Paulo and tap water samples at the terminal sites of the distribution system was sampled repeatedly (22 times) at each of five points in the distribution system; a further 36 samples were taken from cannulae in 19 hemodialysis machines that were ready for the next patient, four samples from the reuse system and 13 from the water storage system. To identify bacteria, samples were filtered through 0.22 µm-pore membranes; for mycobacteria, 0.45 µm pores were used. Conventional microbiological and molecular methods were used in the analysis. Bacteria were isolated from the distribution system (128 isolates), kidney machine water (43) and reuse system (3). Among these isolates, 32 were Gram-positive rods, 120 Gram-negative rods, 20 Gram-positive cocci and 11 mycobacteria. We propose the continual monitoring of the water supplies in hemodialysis centers and the adoption of effective prophylactic measures that minimize the exposure of these immunodeficient patients to contaminated sources of water.

O monitoramento microbiológico da água utilizada no procedimento de hemodiálise é de extrema importância, principalmente devido à imunodebilidade dos pacientes com insuficiência renal crônica. Nosso objetivo foi verificar qualitativa e quantitativamente a presença de bactérias na água de um centro de hemodiálise do interior do Estado de São Paulo. Foram realizadas 22 coletas de cada um dos cinco pontos do sistema de distribuição; 36 amostras de 19 máquinas de hemodiálise, prontas para utilização; quatro amostras do sistema de reuso e 13 amostras do sistema de armazenamento de água, empregando-se a técnica da membrana filtrante com poros de 0,22 µm para bactérias e de 0,45 µm para micobactérias. A identificação foi realizada através de métodos microbiológicos convencionais e de biologia molecular. Isolados bacterianos foram obtidos de sistema de distribuição (128), águas das máquinas (43) e sistema de reuso (3). Entre os isolados 32 foram de bacilos Gram-positivos, 120 bacilos Gram-negativos, 20 Cocos Gram-positivos e 11 micobactérias. Neste estudo, sugerimos que suprimentos de água para o Centro de Hemodiálise devam ser monitorados, adotando-se medidas profiláticas eficazes que minimizem a exposição destes pacientes imunodeficientes a fontes aquáticas ambientais contaminadas.

Determination of the antibacterial activity of crude extracts and compounds isolated from Hortia oreadica (Rutaceae) against oral pathogens.

Extracts from Hortia oreadica afforded four dihydrocinnamic acid derivatives, isolated from the n-hexane extract, as well as limonoid guyanin and the furoquinoline alkaloid dictamnine, both isolated from the dichloromethane extract. The extracts and the isolated compounds were tested against some oral pathogens, so as to investigate their antibacterial activity. The results showed that the n-hexane extract and the compound dictamnine are the most active against the selected microorganisms.

[Isolation of filamentous fungi from water used in a hemodialysis unit]. / Isolamento de fungos filamentosos em água utilizada em uma unidade de hemodiálise.

Despite the relative frequency of opportunistic fungal infections among hemodialysis patients, the reservoirs for these microorganisms in the environment remain unknown, although some recent studies have made correlations with the water supply as their source. The objective of the present study was to monitor the mycological quality of the water system of a hemodialysis unit in the interior of the State of São Paulo, Brazil, over the period from April to July 2006. Fifteen samples of 1000 ml were collected from seven water distribution points using the membrane filtration technique (0.45 microm). A total of 116 filamentous fungus specimens were isolated, including 47 Trichoderma sp (40.5%), 29 Cladosporium sp (25%), 16 Aspergillus sp (13.8%) and 11 Fusarium sp (9.5%). The results suggest that the water supply for hemodialysis units should also be monitored for mycological contamination, and that effective prophylactic measures should be adopted for minimizing the exposure of these immunodeficient patients to contaminated water sources in the environment.

Isolamento de fungos filamentosos em água utilizada em uma unidade de hemodiálise / Isolation of filamentous fungi from water used in a hemodialysis unit

A despeito da relativa freqüência de infecções fúngicas oportunísticas em pacientes sob hemodiálise, os reservatórios ambientais destes permanecem desconhecidos, embora alguns estudos recentes tenham correlacionado o suprimento de água como fonte desses microrganismos. O objetivo deste trabalho foi monitorar a qualidade micológica do sistema hídrico de uma Unidade de Hemodiálise, do interior do Estado de São Paulo, Brasil, no período entre abril e julho de 2006. Foram coletadas amostras (15), de 1000mL em 7 pontos de distribuição de água empregando-se técnica da membrana filtrante (0,45æm). Foram isolados 116 fungos filamentosos, dos quais 47 (40,5 por cento) Trichoderma sp, 29 (25 por cento) Cladosporium sp, 16 (13,8 por cento) Aspergillus sp e 11 (9,5 por cento) Fusarium sp. Mediante os resultados, sugerimos que suprimentos de água para Unidades de Hemodiálise devam ser monitorados também quanto ao aspecto micológico, adotando-se medidas profiláticas eficazes que minimizem a exposição destes pacientes imunodeficientes a fontes aquáticas ambientais contaminadas.

Despite the relative frequency of opportunistic fungal infections among hemodialysis patients, the reservoirs for these microorganisms in the environment remain unknown, although some recent studies have made correlations with the water supply as their source. The objective of the present study was to monitor the mycological quality of the water system of a hemodialysis unit in the interior of the State of São Paulo, Brazil, over the period from April to July 2006. Fifteen samples of 1000 ml were collected from seven water distribution points using the membrane filtration technique (0.45 æm). A total of 116 filamentous fungus specimens were isolated, including 47 Trichoderma sp (40.5 percent), 29 Cladosporium sp (25 percent), 16 Aspergillus sp (13.8 percent) and 11 Fusarium sp (9.5 percent). The results suggest that the water supply for hemodialysis units should also be monitored for mycological contamination, and that effective prophylactic measures should be adopted for minimizing the exposure of these immunodeficient patients to contaminated water sources in the environment.