ESBL in positive hemoculture of a Southern-Brazil Teaching Hospital's Intensive Care Units.

Bloodstream infections are among the most serious and frequent infections, and the people most exposed are patients in the Intensive Care Unit (ICU). ESBL (extended-spectrum beta-lactate) are resistant bacteria to penicillins, cephalosporins and monobactams. It´s necessary to know how often and which microorganisms are involved, checking their susceptibility. This study was carried out at the University Hospital. Data collection was performed in the Adult and Newborn ICUs, with assessment of microorganisms and their resistance profile. During six-month period, 156 samples were studied, and 42 were positive with microorganism isolation. Isolated species include Staphylococcus aureus, Staphylococcus epidermidis and Klebsiella pneumoniae. Many resistant to carbapenem.

Resistance profile and biofilm production of Enterococcus spp., Staphylococcus sp., and Streptococcus spp. from dairy farms in southern Brazil.

Milk is a high nutritional value food that helps in human development and growth. However, it can also harbor microorganisms. Therefore, the objective of this study was to isolate, identify and evaluate the resistance profile and pathogenicity factors of gram-positive cocci isolated from liners in milking rooms in the south of Rio Grande do Sul, Brazil. Biochemical and molecular tests were performed for the identification. The following were isolated: Enterococcus faecalis (10), Enterococcus faecium (4), Staphylococcus intermedius (1), Streptococcus uberis (1), and Streptococcus dysgalactiae (1). The susceptibility of isolated microorganisms to eight antibiotics was evaluated according to CLSI, and the genus that proved to be resistant to most of those was Enterococcus. In addition, all 17 isolates were able to form biofilm, which remained viable after the use of neutral, alkaline and alkaline-chlorinated detergent. The only product that was effective against biofilm of all microorganisms was chlorhexidine 2%. The results obtained highlight the importance of pre- and post-dipping tests on dairy properties, in which chlorhexidine is one of the disinfectants used. As observed, products indicated for cleaning and descaling pipes were not effective on biofilms of the different species tested.

Sensitivity to antifungals by Candida spp samples isolated from cases of chronic atrophic candidiasis (CAC).

The treatment of choice for chronic atrophic candidiasis (CAC), also known as denture stomatitis, is topical antifungal therapy. This study aimed to isolate, identify, and assess the antifungal susceptibility of Candida species from mucosal sites in denture wearers with a diagnosis of CAC and determine the prevalence of associated variables. The sample consisted of 44 patients wearing complete or partial dentures who had a clinical diagnosis of CAC. Using sterile cotton swabs, specimens were collected from the oral mucosa of all patients and grown at 30ºC for 48 h in CHROMagar Candida, as a means of isolating and screening the species. The complementary identification of the species was performed using the VITEK 2 automated system (BioMérieux), as well as the determination of their susceptibility to antifungal agents. Data were analyzed using the chi-square test. STATA 13.1 was used for statistical analysis (α = 5%). Of 44 patients with CAC, 33 (75%) had lesions classified as Newton type II. Yeasts were isolated in 38 cases. The most prevalent species was Candida albicans. None of the isolates were resistant to the antifungals tested. Our findings suggest that current indications for antifungal agents are appropriate. Also, antifungal susceptibility testing and proper fungal identification can help dentists to determine the optimal course of treatment for CAC.

Biochemical identification techniques and antibiotic susceptibility profile of lipolytic ambiental bacteria from effluents.

Different methodologies have been developed throughout the years to identify environmental microorganisms to improve bioremediation techniques, determine susceptibility profiles of bacteria in contaminated environments, and reduce the impact of microorganisms in ecosystems. Two methods of bacterial biochemical identification are compared and the susceptibility profile of bacteria, isolated from residential and industrial wastewater, is determined. Twenty-four bacteria were retrieved from the bacteria bank of the Environmental Microbiology Laboratory at the Institute of Biology (IB) of the Universidade Federal de Pelotas, Pelotas, Brazil. Bacteria were identified by conventional biochemical tests and by the VITEK ®2 automated system. Further, the susceptibility profile to antibiotics was also determined by the automated system. Six species of bacteria (Raoutella planticola, K. pneumoniae ssp. pneumoniae , Serratia marcescens, Raoutella sp., E. cloacae and Klebsiella oxytoca) were identified by conventional biochemical tests, while three species of bacteria (K. pneumoniae ssp. pneumoniae, S. marcescens and K. oxytoca ) were identified by VITEK®2 automated system. VITEK ®2 indicated agreement in 19 (79.17%) isolates and difference in five (20.83%) isolates when compared to results from conventional biochemical tests. Further, antibiotic susceptibility profile results showed that all isolates (100%) were resistant to at least one out of the 18 antibiotics tested by VITEK®2. Thus, no multi-resistant bacteria that may be used in effluent treatment systems or in bioremediation processes have been reported. Results indicate VITEK ® 2 automated system as a potential methodology in the determination of susceptibility profile and identification of environmental bacteria.

Chemical composition of essential oils from the apiaceae family, cytotoxicity, and their antifungal activity in vitro against candida species from oral cavity.

The aims of this research were: evaluate the chemical composition and the cytotoxicity of the Cuminum cyminum (cumin), Anethum graveolens (dill), Pimpinella anisum (anise) and Foeniculum vulgare (fennel) essential oils, as well as their antifungal activity in vitro against ten Candida spp. isolates. The chemical composition of the oils was analyzed by means of gas chromatography coupled with mass spectrometry (GC/MS). The cytotoxicity assays were performed, using the cell proliferation reagent WST-1 in L929 mouse fibroblasts (20x103 well-1). The determinate the Minimum Inhibitory Concentration (MIC), was performed through the Broth Microdilution technique (CLSI). The chemical main components were the cuminaldehyde (32.66%) for cumin, carvone (34.89%) for the dill, trans-anethole (94.01%) for the anise and anethole (79.62%) for the fennel. Anise and fennel did not were cytotoxic in all the tested concentrations, however the cumin oil was cytotoxic in the concentration of 20 mg.mL-1 and the dill in the concentrations of 20 and 8 mg.mL-1. All yeasts were susceptible against the evaluated essential oils. Cumin presented the lowest MIC against yeasts. We concluded that all the essential oils presented inhibitory action against Candida spp., and C . cyminum, P. anisum and F. vulgare were not cytotoxic in the same minimum inhibitory concentrations for the fungi.

ESBL in positive hemoculture of a Southern-Brazil Teaching Hospital's Intensive Care Units / ESBL em hemoculturas positivas das Unidades de Terapia Intensiva de um Hospital Universitário do sul do Brasil

Bloodstream infections are among the most serious and frequent infections, and the people most exposed are patients in the Intensive Care Unit (ICU). ESBL (extended-spectrum beta-lactate) are resistant bacteria to penicillins, cephalosporins and monobactams. It´s necessary to know how often and which microorganisms are involved, checking their susceptibility. This study was carried out at the University Hospital. Data collection was performed in the Adult and Newborn ICUs, with assessment of microorganisms and their resistance profile. During six-month period, 156 samples were studied, and 42 were positive with microorganism isolation. Isolated species include Staphylococcus aureus, Staphylococcus epidermidis and Klebsiella pneumoniae. Many resistant to carbapenem.

ESBL in Positive Hemoculture of a Southern-Brazil Teaching Hospital's Intensive Care Units As infecções da corrente sanguínea estão entre as infecções mais graves e frequentes, e os indivíduos mais expostos são os pacientes da Unidade de Terapia Intensiva (UTI). As ESBL (Beta-Lactamase de Espectro Estendido) são bactérias resistentes a penicilinas, cefalosporinas e monobactâmicos. Se faz necessário saber com que frequência e quais microrganismos estão envolvidos, verificando sua suscetibilidade. Este estudo foi realizado no Hospital Universitário. A coleta de dados foi realizada nas UTIs Adulto e Neonatal, com avaliação dos microrganismos e seu perfil de resistência. Durante o período de seis meses, foram estudadas 156 amostras, sendo 42 positivas com isolamento dos microrganismos. As espécies isoladas incluem Staphylococcus aureus, Staphylococcus epidermidis e Klebsiella pneumoniae. Muitos resistentes aos carbapenêmicos.

Evaluation of the frequency of Candida spp. in hospitalized and non-hospitalized subjects.

The aim of this study was to evaluate the frequency of Candida species between a non-hospitalized and a hospitalized population. For this purpose, samples of saliva were sampled through sterile swabs, moistened in peptone water and rubbed in the oral cavity of 140 individuals, from which, 70 were hospitalized patients from the Medical Clinic of a Teaching Hospital and the other 70 were non-hospitalized subjects. All saliva samples were plated in Sabouraud Dextrose agar added with Chloramphenicol and incubated at 36 °C for 48 hours. The morphology identification was performed through macroscopic and microscopic characterization, the CHROMagar Candida medium and the VITEK® system Yeast Biochemical Card (bio Mérieux SA, France). The results showed a colonization of Candida spp. in 85.7% the hospitalized individuals, where the species found were C. albicans (60%), C. tropicalis (23.4%), C. krusei (3.3%) and Candida spp. (13.3%). In the non-hospitalized individuals the colonization by Candida spp was 47.1%, and the species found were: C. albicans (45.5%), C.krusei (9.1%), C. guilliermondii (9.1% %), C. tropicalis (3.0%), C. famata (3.0%) and Candida spp. (30.3%). In spite of their presence in oral cavity in both groups, Candida spp. was more frequently isolated in hospitalized individuals, who were 6.73 times more likely to have this fungus in the oral cavity and were 3.88 times more likely to have Candida albicans.

Identification and antimicrobial suceptibility profile of bacteria causing bovine mastitis from dairy farms in Pelotas, Rio Grande do Sul.

Mastitis is an inflammatory process of the udder tissue caused mainly by the bacteria Staphylococcus aureus. The indiscriminate use of antibiotics fosters conditions that favor the selection of resistant microorganisms, suppressing at the same time susceptible forms, causing a serious problem in dairy cattle. Given the importance in performing an antibiogram to select the most adequate antimicrobial therapy, the aim of this study was to identify bacteria isolated from cow's milk with mastitis, in dairy farms situated in the city of Pelotas, Rio Grande do Sul, and to determinate the susceptibility profile of these isolates against the antibiotics used to treat this illness. A total of 30 isolates of Staphylococcus spp., were selected from milk samples from the udder quarters with subclinical mastitis whose species were identified through the Vitek system. The susceptibility profile was performed by the disk diffusion assay, against: ampicillin, amoxicillin, bacitracin, cephalexin, ceftiofur, enrofloxacin, gentamicin, neomycin, norfloxacin, penicillin G, tetracycline and trimethoprim. In the antibiogram, 100.0% of the isolates were resistant to trimethoprim and 96.7% to tetracycline and neomycin, three strains of Staphylococcus spp., (10.0%) presented resistance to the 12 antibiotics tested and 24 (80.0%) to at least eight. These results showed the difficulty in treating mastitis, due to the pathogens' resistance.

Antifungal susceptibility profile of diferent yeasts isolates from wild animals, cow's milk with subclinical mastitis and hospital environment.

Yeast infections have acquired great importance due to increasing frequency in immunocompromised patients or patients undergoing invasive diagnostic and therapeutic techniques, and also because of its high morbidity and mortality. At the same time, it has been seen an increase in the emergence of new pathogenic species difficult to diagnose and treat. The aim of this study was to determine the in vitro susceptibility of 89 yeasts from different sources against the antifungals amphotericin B, voriconazole, fluconazole and flucytosine, using the VITEK® 2 Compact system. The antifungal susceptibility was performed automatically by the Vitek® 2 Compact system. The origin of the yeasts was: Group 1 - microbiota of wild animals (W) (26/89), 2 - cow's milk with subclinical mastitis (M) (27/89) and 3 - hospital enviorment (H) (36/89). Of the 89 yeasts submitted to the Vitek® 2 test, 25 (20.9%) were resistant to fluconazole, 11 (12.36%) to amphotericin B, 3 (3.37%) to voriconazole, and no sample was resistant to flucytosine. Regarding the minimum inhibitory concentration (MIC), fluconazole showed an MIC between 1 and 64 mg/mL for the three groups, voriconazole had an MIC between 0.12 and 8 mg/mL, amphotericin B had an MIC between 0.25 and 4 mg/mL for group H and group W respectively, between 0.25 and 16 mg/mL for group M and flucytosine had an MIC equal to 1µg/mL for all groups. The yeasts isolated from the H group showed the highest resistance to fluconazole 12/89 (13.49%), followed by group W (7.87%) and group M (5.62%). The more resistant group to voriconazole was followed by the M and H groups, the W group showed no resistance to this antifungal. Group H was the least resistant (2.25%) to amphotericin.

Airborne fungi in an intensive care unit.

The presence of airborne fungi in Intensive Care Unit (ICUs) is associated with increased nosocomial infections. The aim of this study was the isolation and identification of airborne fungi presented in an ICU from the University Hospital of Pelotas - RS, with the attempt to know the place's environmental microbiota. 40 Petri plates with Sabouraud Dextrose Agar were exposed to an environment of an ICU, where samples were collected in strategic places during morning and afternoon periods for ten days. Seven fungi genera were identified: Penicillium spp. (15.18%), genus with the higher frequency, followed by Aspergillus spp., Cladosporium spp., Fusarium spp., Paecelomyces spp., Curvularia spp., Alternaria spp., Zygomycetes and sterile mycelium. The most predominant fungi genus were Aspergillus spp. (13.92%) in the morning and Cladosporium spp. (13.92%) in the afternoon. Due to their involvement in different diseases, the identified fungi genera can be classified as potential pathogens of inpatients. These results reinforce the need of monitoring the environmental microorganisms with high frequency and efficiently in health institutions.

In vitro susceptibility of nematophagous fungi to antiparasitic drugs: interactions and implications for biological control.

The fast anthelmintic resistance development has shown a limited efficiency in the control of animal's endoparasitosis and has promoted research using alternative control methods. The use of chemicals in animal anthelmintic treatment, in association with nematophagous fungi used for biological control, is a strategy that has proven to be effective in reducing the nematode population density in farm animals. This study aims to verify the in vitro susceptibility of the nematophagous fungi Arthrobotrys oligospora, Duddingtonia flagrans and Paecilomyces lilacinus against the antiparasitic drugs albendazole, thiabendazole, ivermectin, levamisole and closantel by using the Minimum Inhibitory Concentration (MIC). MICs ranged between 4.0 and 0.031 µg/mL for albendazole, thiabendazole and ivermectin, between 0.937 and 0.117 µg/mL for levamisole, and between 0.625 and 0.034 µg/mL for closantel. The results showed that all antiparasitic drugs had an in vitro inhibitory effect on nematophagous fungi, which could compromise their action as agents of biological control. D. flagrans was the most susceptible species to all drugs.

Antiparasitic drugs: in vitro tests against nematophagous fungi.

The use of biological agents has been intensified in recent years against eggs and larvae of gastrointestinal nematodes as an alternative control method in pasture plant health management, with the concomitant use with antiparasitic drugs still occurring. The aim of this study was to test the in vitro activity of the following antiparasitic drugs: Ivermectin and albendazole against the following nematophagous fungi: Paecilomyces fumosoroseus, Paecilomyces lilacinus and Paecilomyces variotii. The agar diffusion test was performed using an initial concentration of 0.0016g/mL of each drug, after solidification of the culture medium containing the drug concentration each nematophagous fungi was inoculated. The results showed that in a concentration of 80µg/mL, the fungal growth decreased, however, with the concentration of 160µg/mL, there was no fungal growth in both drugs, compared to the control, which indicates an inhibition in the development of the nematophagous fungi studied when they come in contact with ivermectin and albendazole.

Sensitivity to antifungals by Candida spp samples isolated from cases of chronic atrophic candidiasis (CAC) / Sensibilidade a antifúngicos por amostras de Candida spp isoladas de casos de candidíase atrófica crônica (CAC)

The treatment of choice for chronic atrophic candidiasis (CAC), also known as denture stomatitis, is topical antifungal therapy. This study aimed to isolate, identify, and assess the antifungal susceptibility of Candida species from mucosal sites in denture wearers with a diagnosis of CAC and determine the prevalence of associated variables. The sample consisted of 44 patients wearing complete or partial dentures who had a clinical diagnosis of CAC. Using sterile cotton swabs, specimens were collected from the oral mucosa of all patients and grown at 30ºC for 48 h in CHROMagar Candida, as a means of isolating and screening the species. The complementary identification of the species was performed using the VITEK 2 automated system (BioMérieux), as well as the determination of their susceptibility to antifungal agents. Data were analyzed using the chi-square test. STATA 13.1 was used for statistical analysis ( = 5%). Of 44 patients with CAC, 33 (75%) had lesions classified as Newton type II. Yeasts were isolated in 38 cases. The most prevalent species was Candida albicans. None of the isolates were resistant to the antifungals tested. Our findings suggest that current indications for antifungal agents are appropriate. Also, antifungal susceptibility testing and proper fungal identification can help dentists to determine the optimal course of treatment for CAC.(AU)

O tratamento de escolha para candidíase atrófica crônica (CAC), também conhecida como estomatite protética, é a terapia antifúngica tópica. Este estudo teve como objetivo isolar, identificar e avaliar a susceptibilidade antifúngica de espécies de Candida de locais mucosos em portadores de prótese com diagnóstico de CAC e determinar a prevalência de variáveis associadas. A amostra consistiu em 44 pacientes portadores de próteses completas ou parciais que tiveram um diagnóstico clínico de CAC. Usando swab estéril, foram coletados espécimes da mucosa oral de todos os pacientes e cultivados a 30ºC durante 48 h em CHROMagar Candida, como forma de isolamento e triagem das espécies. A identificação complementar das espécies foi realizada no sistema automatizado VITEK 2 (BioMérieux), bem como a determinação da susceptibilidade delas a agentes antifúngicos. Os dados foram analisados usando o teste do qui-quadrado. O STATA 13.1 foi utilizado para análise estatística ( = 5%). Dos 44 pacientes com CAC, 33 (75%) apresentaram lesões classificadas como Newton tipo II. As leveduras foram isoladas em 38 casos. A espécie mais prevalente foi Candida albicans. Nenhum dos isolados foi resistente aos antifúngicos testados. Nossas descobertas sugerem que as indicações atuais para os agentes antifúngicos são apropriadas. Além disso, testes de susceptibilidade antifúngicos e identificação fúngica adequada podem ajudar os dentistas a determinar o curso ótimo de tratamento para CAC.(AU)

Sensitivity to antifungals by Candida spp samples isolated from cases of chronic atrophic candidiasis (CAC) / Sensibilidade a antifúngicos por amostras de Candida spp isoladas de casos de candidíase atrófica crônica (CAC)

Abstract The treatment of choice for chronic atrophic candidiasis (CAC), also known as denture stomatitis, is topical antifungal therapy. This study aimed to isolate, identify, and assess the antifungal susceptibility of Candida species from mucosal sites in denture wearers with a diagnosis of CAC and determine the prevalence of associated variables. The sample consisted of 44 patients wearing complete or partial dentures who had a clinical diagnosis of CAC. Using sterile cotton swabs, specimens were collected from the oral mucosa of all patients and grown at 30ºC for 48 h in CHROMagar Candida, as a means of isolating and screening the species. The complementary identification of the species was performed using the VITEK 2 automated system (BioMérieux), as well as the determination of their susceptibility to antifungal agents. Data were analyzed using the chi-square test. STATA 13.1 was used for statistical analysis (α = 5%). Of 44 patients with CAC, 33 (75%) had lesions classified as Newton type II. Yeasts were isolated in 38 cases. The most prevalent species was Candida albicans. None of the isolates were resistant to the antifungals tested. Our findings suggest that current indications for antifungal agents are appropriate. Also, antifungal susceptibility testing and proper fungal identification can help dentists to determine the optimal course of treatment for CAC.

Resumo O tratamento de escolha para candidíase atrófica crônica (CAC), também conhecida como estomatite protética, é a terapia antifúngica tópica. Este estudo teve como objetivo isolar, identificar e avaliar a susceptibilidade antifúngica de espécies de Candida de locais mucosos em portadores de prótese com diagnóstico de CAC e determinar a prevalência de variáveis associadas. A amostra consistiu em 44 pacientes portadores de próteses completas ou parciais que tiveram um diagnóstico clínico de CAC. Usando swab estéril, foram coletados espécimes da mucosa oral de todos os pacientes e cultivados a 30ºC durante 48 h em CHROMagar Candida, como forma de isolamento e triagem das espécies. A identificação complementar das espécies foi realizada no sistema automatizado VITEK 2 (BioMérieux), bem como a determinação da susceptibilidade delas a agentes antifúngicos. Os dados foram analisados usando o teste do qui-quadrado. O STATA 13.1 foi utilizado para análise estatística (α = 5%). Dos 44 pacientes com CAC, 33 (75%) apresentaram lesões classificadas como Newton tipo II. As leveduras foram isoladas em 38 casos. A espécie mais prevalente foi Candida albicans. Nenhum dos isolados foi resistente aos antifúngicos testados. Nossas descobertas sugerem que as indicações atuais para os agentes antifúngicos são apropriadas. Além disso, testes de susceptibilidade antifúngicos e identificação fúngica adequada podem ajudar os dentistas a determinar o curso ótimo de tratamento para CAC.

Sensitivity to antifungals by Candida spp samples isolated from cases of chronic atrophic candidiasis (CAC)

Abstract The treatment of choice for chronic atrophic candidiasis (CAC), also known as denture stomatitis, is topical antifungal therapy. This study aimed to isolate, identify, and assess the antifungal susceptibility of Candida species from mucosal sites in denture wearers with a diagnosis of CAC and determine the prevalence of associated variables. The sample consisted of 44 patients wearing complete or partial dentures who had a clinical diagnosis of CAC. Using sterile cotton swabs, specimens were collected from the oral mucosa of all patients and grown at 30ºC for 48 h in CHROMagar Candida, as a means of isolating and screening the species. The complementary identification of the species was performed using the VITEK 2 automated system (BioMérieux), as well as the determination of their susceptibility to antifungal agents. Data were analyzed using the chi-square test. STATA 13.1 was used for statistical analysis ( = 5%). Of 44 patients with CAC, 33 (75%) had lesions classified as Newton type II. Yeasts were isolated in 38 cases. The most prevalent species was Candida albicans. None of the isolates were resistant to the antifungals tested. Our findings suggest that current indications for antifungal agents are appropriate. Also, antifungal susceptibility testing and proper fungal identification can help dentists to determine the optimal course of treatment for CAC.

Resumo O tratamento de escolha para candidíase atrófica crônica (CAC), também conhecida como estomatite protética, é a terapia antifúngica tópica. Este estudo teve como objetivo isolar, identificar e avaliar a susceptibilidade antifúngica de espécies de Candida de locais mucosos em portadores de prótese com diagnóstico de CAC e determinar a prevalência de variáveis associadas. A amostra consistiu em 44 pacientes portadores de próteses completas ou parciais que tiveram um diagnóstico clínico de CAC. Usando swab estéril, foram coletados espécimes da mucosa oral de todos os pacientes e cultivados a 30ºC durante 48 h em CHROMagar Candida, como forma de isolamento e triagem das espécies. A identificação complementar das espécies foi realizada no sistema automatizado VITEK 2 (BioMérieux), bem como a determinação da susceptibilidade delas a agentes antifúngicos. Os dados foram analisados usando o teste do qui-quadrado. O STATA 13.1 foi utilizado para análise estatística ( = 5%). Dos 44 pacientes com CAC, 33 (75%) apresentaram lesões classificadas como Newton tipo II. As leveduras foram isoladas em 38 casos. A espécie mais prevalente foi Candida albicans. Nenhum dos isolados foi resistente aos antifúngicos testados. Nossas descobertas sugerem que as indicações atuais para os agentes antifúngicos são apropriadas. Além disso, testes de susceptibilidade antifúngicos e identificação fúngica adequada podem ajudar os dentistas a determinar o curso ótimo de tratamento para CAC.

Chemical composition of essential oils from the apiaceae family, cytotoxicity, and their antifungal activity in vitro against candida species from oral cavity / Composição química de óleos essenciais da família apiaceae, citotoxicidade e sua atividade antifúngica in vitro contra espécies de candida da cavidade oral

The aims of this research were: evaluate the chemical composition and the cytotoxicity of the Cuminum cyminum (cumin), Anethum graveolens (dill), Pimpinella anisum (anise) and Foeniculum vulgare (fennel) essential oils, as well as their antifungal activity in vitro against ten Candida spp. isolates. The chemical composition of the oils was analyzed by means of gas chromatography coupled with mass spectrometry (GC/MS). The cytotoxicity assays were performed, using the cell proliferation reagent WST-1 in L929 mouse fibroblasts (20x103 well-1). The determinate the Minimum Inhibitory Concentration (MIC), was performed through the Broth Microdilution technique (CLSI). The chemical main components were the cuminaldehyde (32.66%) for cumin, carvone (34.89%) for the dill, trans-anethole (94.01%) for the anise and anethole (79.62%) for the fennel. Anise and fennel did not were cytotoxic in all the tested concentrations, however the cumin oil was cytotoxic in the concentration of 20 mg.mL-1 and the dill in the concentrations of 20 and 8 mg.mL-1. All yeasts were susceptible against the evaluated essential oils. Cumin presented the lowest MIC against yeasts. We concluded that all the essential oils presented inhibitory action against Candida spp., and C . cyminum, P. anisum and F. vulgare were not cytotoxic in the same minimum inhibitory concentrations for the fungi.(AU)

Os objetivos desta pesquisa foram: avaliar a composição química e a citotoxicidade dos óleos essenciais de Cuminum cyminum (cominho), Anethum graveolens (endro), Pimpinella anisum (erva-doce) e Foeniculum vulgare (funcho), bem como sua atividade antifúngica in vitro contra dez isolados de Candida spp.. A composição química dos óleos foi analisada por meio de cromatografia gasosa acoplada à espectrometria de massa (GC / MS). Os ensaios de citotoxicidade foram realizados, utilizando o reagente de proliferação celular WST-1 em fibroblastos de ratinho L929 (20x103 poço-1). A determinação da Concentração Inibitória Mínima (MIC) foi realizada através da técnica de microdiluição em caldo (CLSI). Os principais componentes químicos foram o cuminaldeído (32.66%) para cominho, carvona (34.89%) para o endro, trans-anetol (94.01%) para erva-doce e anetol (79.62%) para a funcho. O endro e a erva-doce não foram citotóxicos em todas as concentrações testadas, no entanto, o óleo de cominho foi citotóxico na concentração de 20 mg.mL-1 e o endro nas concentrações de 20 e 8 mg.mL-1. Todas as leveduras foram suscetíveis aos óleos essenciais avaliados. O cominho apresentou a menor CIM contra as leveduras. Concluímos que todos os óleos essenciais apresentaram ação inibidora contra Candida spp., e C. cyminum, P. anisum e F. vulgare não foram citotóxicos nas mesmas concentrações inibitórias mínimas para os fungos.(AU)

Biochemical identification techniques and antibiotic susceptibility profile of lipolytic ambiental bacteria from effluents / Técnicas de identificação bioquímica e perfil de suscetibilidade antibiótica de bactérias ambientais lipolíticas de efluentes

Different methodologies have been developed throughout the years to identify environmental microorganisms to improve bioremediation techniques, determine susceptibility profiles of bacteria in contaminated environments, and reduce the impact of microorganisms in ecosystems. Two methods of bacterial biochemical identification are compared and the susceptibility profile of bacteria, isolated from residential and industrial wastewater, is determined. Twenty-four bacteria were retrieved from the bacteria bank of the Environmental Microbiology Laboratory at the Institute of Biology (IB) of the Universidade Federal de Pelotas, Pelotas, Brazil. Bacteria were identified by conventional biochemical tests and by the VITEK ®2 automated system. Further, the susceptibility profile to antibiotics was also determined by the automated system. Six species of bacteria (Raoutella planticola, K. pneumoniae ssp. pneumoniae , Serratia marcescens, Raoutella sp., E. cloacae and Klebsiella oxytoca) were identified by conventional biochemical tests, while three species of bacteria (K. pneumoniae ssp. pneumoniae, S. marcescens and K. oxytoca ) were identified by VITEK®2 automated system. VITEK ®2 indicated agreement in 19 (79.17%) isolates and difference in five (20.83%) isolates when compared to results from conventional biochemical tests. Further, antibiotic susceptibility profile results showed that all isolates (100%) were resistant to at least one out of the 18 antibiotics tested by VITEK®2. Thus, no multi-resistant bacteria that may be used in effluent treatment systems or in bioremediation processes have been reported. Results indicate VITEK ® 2 automated system as a potential methodology in the determination of susceptibility profile and identification of environmental bacteria.(AU)

Diferentes metodologias foram desenvolvidas ao longo dos anos para identificar microrganismos ambientais para melhorar as técnicas de biorremediação, determinar perfis de suscetibilidade de bactérias em ambientes contaminados e reduzir o impacto de microrganismos nos ecossistemas. Dois métodos de identificação bioquímica bacteriana são comparados e o perfil de susceptibilidade de bactérias, isoladas de efluentes residenciais e industriais, é determinado. Vinte e quatro bactérias foram coletadas do banco de bactérias do Laboratório de Microbiologia Ambiental do Instituto de Biologia (IB) da Universidade Federal de Pelotas, Pelotas, Brasil. As bactérias foram identificadas por testes bioquímicos convencionais e pelo sistema automatizado VITEK®2. Além disso, o perfil de suscetibilidade aos antibióticos também foi determinado pelo sistema automatizado. Seis espécies de bactérias (Raoutella planticola , K. pneumoniae ssp. pneumoniae, Serratia marcescens, Raoutella sp., E. cloacae e Klebsiella oxytoca) foram identificadas por testes bioquímicos convencionais, enquanto três espécies de bactérias (K. pneumoniae ssp. pneumoniae, S. marcescens e K. oxytoca) foram identificados pelo sistema automatizado VITEK®2. VITEK®2 indicou concordância em 19 (79,17%) isolados e diferença em cinco (20,83%) isolados quando comparados aos resultados de testes bioquímicos convencionais. Além disso, os resultados do perfil de suscetibilidade aos antibióticos mostraram que todos os isolados (100%) foram resistentes a pelo menos um dos 18 antibióticos testados pelo VITEK®2. Assim, não foram relatadas bactérias multirresistentes que possam ser usadas em sistemas de tratamento de efluentes ou em processos de biorremediação. Os resultados indicam que o sistema automatizado VITEK ® 2 é uma metodologia potencial na determinação do perfil de suscetibilidade e identificação de bactérias ambientais.(AU)

Chemical composition of essential oils from the apiaceae family, cytotoxicity, and their antifungal activity in vitro against candida species from oral cavity / Composição química de óleos essenciais da família apiaceae, citotoxicidade e sua atividade antifúngica in vitro contra espécies de candida da cavidade oral

Abstract The aims of this research were: evaluate the chemical composition and the cytotoxicity of the Cuminum cyminum (cumin), Anethum graveolens (dill), Pimpinella anisum (anise) and Foeniculum vulgare (fennel) essential oils, as well as their antifungal activity in vitro against ten Candida spp. isolates. The chemical composition of the oils was analyzed by means of gas chromatography coupled with mass spectrometry (GC/MS). The cytotoxicity assays were performed, using the cell proliferation reagent WST-1 in L929 mouse fibroblasts (20x103 well-1). The determinate the Minimum Inhibitory Concentration (MIC), was performed through the Broth Microdilution technique (CLSI). The chemical main components were the cuminaldehyde (32.66%) for cumin, carvone (34.89%) for the dill, trans-anethole (94.01%) for the anise and anethole (79.62%) for the fennel. Anise and fennel did not were cytotoxic in all the tested concentrations, however the cumin oil was cytotoxic in the concentration of 20 mg.mL-1 and the dill in the concentrations of 20 and 8 mg.mL-1. All yeasts were susceptible against the evaluated essential oils. Cumin presented the lowest MIC against yeasts. We concluded that all the essential oils presented inhibitory action against Candida spp., and C . cyminum, P. anisum and F. vulgare were not cytotoxic in the same minimum inhibitory concentrations for the fungi.

Resumo Os objetivos desta pesquisa foram: avaliar a composição química e a citotoxicidade dos óleos essenciais de Cuminum cyminum (cominho), Anethum graveolens (endro), Pimpinella anisum (erva-doce) e Foeniculum vulgare (funcho), bem como sua atividade antifúngica in vitro contra dez isolados de Candida spp.. A composição química dos óleos foi analisada por meio de cromatografia gasosa acoplada à espectrometria de massa (GC / MS). Os ensaios de citotoxicidade foram realizados, utilizando o reagente de proliferação celular WST-1 em fibroblastos de ratinho L929 (20x103 poço-1). A determinação da Concentração Inibitória Mínima (MIC) foi realizada através da técnica de microdiluição em caldo (CLSI). Os principais componentes químicos foram o cuminaldeído (32.66%) para cominho, carvona (34.89%) para o endro, trans-anetol (94.01%) para erva-doce e anetol (79.62%) para a funcho. O endro e a erva-doce não foram citotóxicos em todas as concentrações testadas, no entanto, o óleo de cominho foi citotóxico na concentração de 20 mg.mL-1 e o endro nas concentrações de 20 e 8 mg.mL-1. Todas as leveduras foram suscetíveis aos óleos essenciais avaliados. O cominho apresentou a menor CIM contra as leveduras. Concluímos que todos os óleos essenciais apresentaram ação inibidora contra Candida spp., e C. cyminum, P. anisum e F. vulgare não foram citotóxicos nas mesmas concentrações inibitórias mínimas para os fungos.

Biochemical identification techniques and antibiotic susceptibility profile of lipolytic ambiental bacteria from effluents / Técnicas de identificação bioquímica e perfil de suscetibilidade antibiótica de bactérias ambientais lipolíticas de efluentes

Abstract Different methodologies have been developed throughout the years to identify environmental microorganisms to improve bioremediation techniques, determine susceptibility profiles of bacteria in contaminated environments, and reduce the impact of microorganisms in ecosystems. Two methods of bacterial biochemical identification are compared and the susceptibility profile of bacteria, isolated from residential and industrial wastewater, is determined. Twenty-four bacteria were retrieved from the bacteria bank of the Environmental Microbiology Laboratory at the Institute of Biology (IB) of the Universidade Federal de Pelotas, Pelotas, Brazil. Bacteria were identified by conventional biochemical tests and by the VITEK ®2 automated system. Further, the susceptibility profile to antibiotics was also determined by the automated system. Six species of bacteria (Raoutella planticola, K. pneumoniae ssp. pneumoniae , Serratia marcescens, Raoutella sp., E. cloacae and Klebsiella oxytoca) were identified by conventional biochemical tests, while three species of bacteria (K. pneumoniae ssp. pneumoniae, S. marcescens and K. oxytoca ) were identified by VITEK®2 automated system. VITEK ®2 indicated agreement in 19 (79.17%) isolates and difference in five (20.83%) isolates when compared to results from conventional biochemical tests. Further, antibiotic susceptibility profile results showed that all isolates (100%) were resistant to at least one out of the 18 antibiotics tested by VITEK®2. Thus, no multi-resistant bacteria that may be used in effluent treatment systems or in bioremediation processes have been reported. Results indicate VITEK ® 2 automated system as a potential methodology in the determination of susceptibility profile and identification of environmental bacteria.

Resumo Diferentes metodologias foram desenvolvidas ao longo dos anos para identificar microrganismos ambientais para melhorar as técnicas de biorremediação, determinar perfis de suscetibilidade de bactérias em ambientes contaminados e reduzir o impacto de microrganismos nos ecossistemas. Dois métodos de identificação bioquímica bacteriana são comparados e o perfil de susceptibilidade de bactérias, isoladas de efluentes residenciais e industriais, é determinado. Vinte e quatro bactérias foram coletadas do banco de bactérias do Laboratório de Microbiologia Ambiental do Instituto de Biologia (IB) da Universidade Federal de Pelotas, Pelotas, Brasil. As bactérias foram identificadas por testes bioquímicos convencionais e pelo sistema automatizado VITEK®2. Além disso, o perfil de suscetibilidade aos antibióticos também foi determinado pelo sistema automatizado. Seis espécies de bactérias (Raoutella planticola , K. pneumoniae ssp. pneumoniae, Serratia marcescens, Raoutella sp., E. cloacae e Klebsiella oxytoca) foram identificadas por testes bioquímicos convencionais, enquanto três espécies de bactérias (K. pneumoniae ssp. pneumoniae, S. marcescens e K. oxytoca) foram identificados pelo sistema automatizado VITEK®2. VITEK®2 indicou concordância em 19 (79,17%) isolados e diferença em cinco (20,83%) isolados quando comparados aos resultados de testes bioquímicos convencionais. Além disso, os resultados do perfil de suscetibilidade aos antibióticos mostraram que todos os isolados (100%) foram resistentes a pelo menos um dos 18 antibióticos testados pelo VITEK®2. Assim, não foram relatadas bactérias multirresistentes que possam ser usadas em sistemas de tratamento de efluentes ou em processos de biorremediação. Os resultados indicam que o sistema automatizado VITEK ® 2 é uma metodologia potencial na determinação do perfil de suscetibilidade e identificação de bactérias ambientais.

Identification and antimicrobial suceptibility profile of bacteria causing bovine mastitis from dairy farms in Pelotas, Rio Grande do Sul / Identificação e perfil de suscetibilidade antimicrobiana de bactérias causadoras de mastite bovina em propriedades leiteiras de Pelotas, Rio Grande do Sul

Mastitis is an inflammatory process of the udder tissue caused mainly by the bacteria Staphylococcus aureus. The indiscriminate use of antibiotics fosters conditions that favor the selection of resistant microorganisms, suppressing at the same time susceptible forms, causing a serious problem in dairy cattle. Given the importance in performing an antibiogram to select the most adequate antimicrobial therapy, the aim of this study was to identify bacteria isolated from cows milk with mastitis, in dairy farms situated in the city of Pelotas, Rio Grande do Sul, and to determinate the susceptibility profile of these isolates against the antibiotics used to treat this illness. A total of 30 isolates of Staphylococcus spp., were selected from milk samples from the udder quarters with subclinical mastitis whose species were identified through the Vitek system. The susceptibility profile was performed by the disk diffusion assay, against: ampicillin, amoxicillin, bacitracin, cephalexin, ceftiofur, enrofloxacin, gentamicin, neomycin, norfloxacin, penicillin G, tetracycline and trimethoprim. In the antibiogram, 100.0% of the isolates were resistant to trimethoprim and 96.7% to tetracycline and neomycin, three strains of Staphylococcus spp., (10.0%) presented resistance to the 12 antibiotics tested and 24 (80.0%) to at least eight. These results showed the difficulty in treating mastitis, due to the pathogens resistance.(AU)

A mastite se constitui no processo inflamatório da glândula mamária causada principalmente por bactérias Staphylococcus aureus. O uso indiscriminado dos antibióticos promove condições que favorecem a seleção de micro-organismos resistentes e, ao mesmo tempo, suprime formas suscetíveis, causando um grave problema para a bovinocultura leiteira. Tendo em vista a importância da realização do antibiograma para a seleção da terapia antimicrobiana mais adequada, o objetivo deste estudo foi identificar bactérias isoladas de leite de vaca com mastite, oriundas de propriedades leiteiras localizadas na cidade de Pelotas, RS, bem como determinar o perfil de suscetibilidade desses isolados frente a antibióticos usados para o tratamento desta doença. Foram selecionados 30 isolados de Staphylococcus spp. de amostras de leite provenientes de quartos mamários com mastite subclínica, cujas espécies foram identificadas através do sistema Vitek. O perfil de suscetibilidade foi realizado pela técnica de difusão em disco, frente a: ampicilina, amoxicilina, bacitracina, cefalexina, ceftiofur, enrofloxacina, gentamicina, neomicina, norfloxacina, penicilina G, tetraciclina e trimetoprima. No antibiograma, 100,0% dos isolados foram resistentes a trimetoprima e 96,7% a tetraciclina e a neomicina, três cepas (10,0%) foram resistentes aos 12 antibióticos testados e 24 (80,0%) a pelo menos oito. Esses resultados demonstram a dificuldade encontrada no tratamento da mastite devido à resistência dos agentes patológicos.(AU)