Detec����o de Enterococcus resistentes a vancomicina em cria����es comerciais de ovinos e caprinos das regi��es centro-leste e nordeste do Estado de S��o Paulo

As exig��ncias das condi����es higi��nico-sanit��rias na produ����o de animais de interesse zoot��cnico v��m aumentando progressivamente dada �� necessidade de aliar-se produtividade a produtos de alta qualidade para atender a mercados consumidores cada vez mais exigentes. Nesse sentido, a utiliza����o de antimicrobianos, tanto na profilaxia como na terap��utica, permanece como estrat��gia de controle para v��rios microrganismos patog��nicos, de import��ncia n��o apenas para a produ����o animal como tamb��m para a sa��de humana, ainda que restri����es ao uso indiscriminado desses produtos t��m se intensificado. N��o obstante, o uso excessivo desses produtos est�� associado �� sele����o de microrganismos resistentes nas ��reas de produ����o. Por outro lado, investiga����es sobre circula����o de cepas resistentes em rebanhos animais, at�� ent��o restritas a popula����es humanas, ainda permanecem limitadas no Brasil. Bact��rias do g��nero Enterococcus, integrantes usuais da microbiota gastrointestinal animal e humana, s��o indicadoras ambientais de contamina����o fecal e tem-se tornado objeto de preocupa����o em sa��de p��blica e veterin��ria dada a ocorr��ncia de cepas resistentes �� vancomicina (VRE). O presente trabalho teve como objetivo isolar, quantificar e caracterizar VRE presentes em amostras fecais de ovinos oriundos de pequenas propriedades das regi��es centro-leste e nordeste do estado de S��o Paulo. Para tanto, 132 amostras fecais foram coletadas diretamente do reto dos animais ou do piso das instala����es. As amostras foram semeadas em ��gar m-Enterococcus e subcultivadas em ��gar Bile Esculina acrescido de 6 µg/mL de vancomicina (ABEV), para confirma����o de Enterococcus spp e detec����o de cepas resistentes. Procedeu-se igualmente a observa����o da morfologia, caracter��sticas tintoriais, bioqu��micas e moleculares. O n��mero m��ximo de Enterococcus spp. encontrado foi de 2,6 �� 105 e 1,70 �� 105 UFC/g de fezes do ambiente e dos animais, respectivamente. Na caracteriza����o bioqu��mica esp��cies mais prevalentes foram: Enterococcus faecalis e Vagococcus fluvialis. No ABEV, houve crescimento de col��nias VRE em 33 das 84 amostras de ovinos-caprinos e em 21 das 48 amostras ambientais, representando, respectivamente 46,7% e 29,3% das amostras analisadas. A an��lise por multiplex PCR das 54 cepas VRE obtidas indicaram que 23 (43%), 22 (41%), 2 (3,5%) e 2 (3,5%) foram positivas, respectivamente, para os genes vanC2/C3, vanC1, vanA e vanB, sendo que para 5,3% dos isolados nenhum produto foi amplificado, sugerindo a poss��vel ocorr��ncia de genes dos demais grupos van conhecidos entre os isolados. Os resultados obtidos indicam, de forma in��dita no pa��s, a circula����o de VRE em propriedades produtoras de ovinos e caprinos, sem ocorr��ncia de manifesta����es cl��nicas aparentes nos animais, por��m com poss��veis riscos �� sa��de dos produtores e profissionais envolvidos, bem como a eventuais consumidores.

Superf��cies do ambiente hospitalar como poss��veis reservat��rios de bact��rias resistentes: uma revis��o

O principal objetivo deste estudo �� identificar, na literatura, artigos sobre a ocorr��ncia de contamina����o das superf��cies inanimadas e uma poss��vel dissemina����o de bact��rias resistentes no ambiente hospitalar. Realizou-se um levantamento bibliogr��fico de artigos publicados nas bases de dados LILACS, MEDLINE, Science Direct, SCOPUS e ISI Web of Knowledge, entre 2000 e 2008. Foram selecionados e analisados vinte e um artigos. Nos estudos analisados, real��ou-se a presen��a de bact��rias em monitores, grades de cama, mesas, torneiras, telefones, teclados de computador e outros objetos. Houve predomin��ncia de Staphylococcus aureus resistente �� meticilina, Clostridium difficile, Acine-to-bacter baumannii e Enterococcus resistentes �� vancomicina, sendo fator preditivo a ocupa����o pr��via por pacientes colonizados por tais microrganismos. Verificou-se semelhan��a entre as cepas isoladas de pacientes colonizados e/ou infectados e as cepas do ambiente por tipifica����o molecular. Essas evid��ncias refor��am a necessidade de conhecimento e controle de fontes de pat��genos no ambiente hospitalar.

Epidemiologia molecular, fatores de risco e progn��stico da infec����o por Enterococcus spp resistentes �� vancomicina em situa����o de introdu����o recente: um estudo em dois hospitais p��blicos de Bauru-SP

P��s-gradua����o em Doen��as Tropicais - FMB

Incid��ncia de Enterococcus resistente �� vancomicina em hospital universit��rio no Brasil

OBJETIVO: O enterococo resistente �� vancomicina �� atualmente um dos principais microorganismos implicados em infec����es nosocomiais. Assim, realizou-se estudo com o objetivo de avaliar sua epidemiologia em um hospital terci��rio de ensino. M��TODOS: Trata-se de um estudo epidemiol��gico retrospectivo, realizado de 2000 a 2002, que analisou amostras de culturas cl��nicas positivas para enterococo resistente �� vancomicina (VRE) em um hospital universit��rio com 660 leitos. Procurou-se definir sua incid��ncia e os principais s��tios e unidades de isolamento. Foi verificada a signific��ncia entre as vari��veis nos tr��s anos de estudo, sendo considerado como significante p<0,05. RESULTADOS: Houve aumento progressivo na resist��ncia �� vancomicina nas culturas cl��nicas positivas para Enterococcus spp. nos tr��s anos de estudo. Em 2000, 9,5% das amostras eram resistentes �� vancomicina, com aumento para 14,7% em 2001 e 15,8% em 2002. As unidades com maior n��mero de isolados foram respectivamente: pronto-socorro (19,5%) e UTI geral (15%); os s��tios mais isolados foram: urina (36%) e sangue (20%). CONCLUS��ES: Com o aumento progressivo na incid��ncia de resist��ncia �� vancomicina e da taxa de VRE, concluiu-se ser necess��rias medidas de controle mais efetivas para deter a dissemina����o do VRE.

Estudo da actividade de lisinas codificadas por bacteri��fagos que infectam Enterococcus sp

Disserta����o apresentada para a obten����o do Grau de Mestre em Gen��tica Molecular e Biomedicina, pela Universidade Nova de Lisboa, Faculdade de Ci��ncias e Tecnologia

Resist��ncia antimicrobiana associada em isolados cl��nicos de Enterococcus spp

INTRODU����O: O aumento da preval��ncia de isolados de enterococos em hospitais, particularmente Enterococcus resistente �� vancomicina (VRE), �� importante por causa da limitada terapia antimicrobiana efetiva para o tratamento de infec����es enteroc��cicas. M��TODOS: O presente trabalho apresentou uma investiga����o retrospectiva de dados de suscetibilidade in vitro quantitativa para uma variedade de antimicrobianos frente aos isolados de Enterococcus spp. e avalia����o da associa����o de resist��ncia entre os agentes antimicrobianos apontados como escolha para o tratamento de infec����es causadas por VRE, atrav��s do c��lculo do risco relativo. RESULTADOS: Dos 156 isolados de enterococos, 40 (25,6%) foram resistentes a tr��s ou mais antimicrobianos, incluindo 7,7% (n = 12/156) resistentes �� vancomicina. A associa����o de resist��ncia elevada foi mais pronunciada entre os isolados de VREs com antimicrobianos alternativos e prim��rios para o tratamento de infec����es causadas por estes pat��genos, incluindo ampicilina (100%, RR = 7,2), estreptomicina (90,9%, RR = 4,9), rifampicina (91,7%, RR = 3,1) e linezolida (50%, RR = 11,5), apesar da alta taxa de suscetibilidade a esta droga (94,9%). CONCLUS��ES: A resist��ncia associada significativa aos antimicrobianos de primeira escolha e alternativos, usados no tratamento de infec����es graves por cepas com o fen��tipo VRE e que requerem um regime terap��utico combinado, evidencia alternativas terap��uticas ainda mais limitadas na institui����o analisada.

Resist��ncia antimicrobiana em Enterococcus faecalis e Enterococcus faecium isolados de carca��as de frango

O objetivo deste trabalho foi realizar o isolamento e analisar o perfil de resist��ncia antimicrobiana de Enterococcus de carca��as de frango resfriadas e congeladas comercializadas no Distrito Federal, detectando genes de resist��ncia antimicrobiana e identificando as esp��cies Enterococcus faecalis e Enterococcus faecium por rea����o polimerase em cadeia. Foram analisadas 100 carca��as de frangos, das quais foram isoladas 50 cepas de Enterococcus spp., sendo 42% de E. faecalis e 2% de E. faecium. O teste de susceptibilidade antimicrobiana demonstrou que todas as cepas isoladas apresentaram resist��ncia a pelo menos um antimicrobiano, dos quais 90,47% das cepas de E. faecalis, 100% das cepas de E. Faecium e 82,14% dos Enterococcus spp. apresentaram resist��ncia �� Tetraciclina; 80,95% das cepas de E. faecalis e 35,71% das cepas de Enterococcus spp. foram resistentes �� Eritromicina; 39,28% dos Enterococcus spp. e 23,80% dos E. faecalis �� Ciprofloxacina e 28,57% dos E. faecalis apresentaram resist��ncia ao Cloranfenicol. Foram detectados os genes de resist��ncia antimicrobiana erm(B), vanC-1, aph(3')-llla, ant(6)-la, vanB, vanA, aac(6')-le-aph(2'')-la, erm(A) e tet(M) - este ��ltimo mais frequente. Estes resultados sugerem s��rios problemas para a Sa��de P��blica, uma vez que esses microrganismos podem possuir a capacidade de transmitir genes de resist��ncia antimicrobiana para outros microrganismos presentes na microbiota intestinal de humanos e animais, podendo inviabilizar o uso destas drogas para tratamentos cl��nicos.

Preval��ncia e suscetibilidade aos antibi��ticos de enterococcus spp. isoladas da cavidade bucal humana

Coordena����o de Aperfei��oamento de Pessoal de N��vel Superior (CAPES)

Preval��ncia, Virul��ncia e sensibilidade ��s terapias antimicrobianas das cepas de enterecoccus faecalis e enterococcus faecium isoladas de infec����es endod��nticas

P��s-gradua����o em Biopatologia Bucal - ICT

Evaluation Of Photodynamic Therapy Using A Diode Laser And Different Photosensitizers Against Enterococcus Faecalis.

Photodynamic therapy (PDT) has been proven to be effective in disinfecting root canals. The aim of this present study was to evaluate the effects of PDT on the viability of Enterococcus faecalis using methylene blue (MB) and malachite green (MG) as photosensitizers. Solutions containing E. faecalis (ATCC 29212) were prepared and harvested by centrifugation to obtain cell suspensions, which were mixed with MB and MG. Samples were individually irradiated by the diode laser at a distance of 1mm for 30, 60, or 120 seconds. Colonyforming units (CFU) were determined for each treatment. PDT for 60 and 120 seconds with MG reduced E. faecalis viability significantly. Similar results were obtained when MB was used as photosensitizer. PDT using MB and MG have antibacterial effect against E. faecalis, showing potential to be used as an adjunctive antimicrobial procedure in endodontic therapy.

Behavior Of Listeria Monocytogenes In A Multi-species Biofilm With Enterococcus Faecalis And Enterococcus Faecium And Control Through Sanitation Procedures.

The formation of mono-species biofilm (Listeria monocytogenes) and multi-species biofilms (Enterococcus faecium, Enterococcus faecalis, and L. monocytogenes) was evaluated. In addition, the effectiveness of sanitation procedures for the control of the multi-species biofilm also was evaluated. The biofilms were grown on stainless steel coupons at various incubation temperatures (7, 25 and 39��C) and contact times (0, 1, 2, 4, 6 and 8days). In all tests, at 7��C, the microbial counts were below 0.4 log CFU/cm(2) and not characteristic of biofilms. In mono-species biofilm, the counts of L. monocytogenes after 8days of contact were 4.1 and 2.8 log CFU/cm(2) at 25 and 39��C, respectively. In the multi-species biofilms, Enterococcus spp. were present at counts of 8 log CFU/cm(2) at 25 and 39��C after 8days of contact. However, the L. monocytogenes in multi-species biofilms was significantly affected by the presence of Enterococcus spp. and by temperature. At 25��C, the growth of L. monocytogenes biofilms was favored in multi-species cultures, with counts above 6 log CFU/cm(2) after 8days of contact. In contrast, at 39��C, a negative effect was observed for L. monocytogenes biofilm growth in mixed cultures, with a significant reduction in counts over time and values below 0.4 log CFU/cm(2) starting at day 4. Anionic tensioactive cleaning complemented with another procedure (acid cleaning, disinfection or acid cleaning+disinfection) eliminated the multi-species biofilms under all conditions tested (counts of all micro-organisms<0.4 log CFU/cm(2)). Peracetic acid was the most effective disinfectant, eliminating the multi-species biofilms under all tested conditions (counts of the all microorganisms <0.4 log CFU/cm(2)). In contrast, biguanide was the least effective disinfectant, failing to eliminate biofilms under all the test conditions.

Biofilms Of Enterococcus Faecalis And Enterococcus Faecium Isolated From The Processing Of Ricotta And The Control Of These Pathogens Through Cleaning And Sanitization Procedures.

The biofilm formation of Enterococcus faecalis and Enterococcus faecium isolated from the processing of ricotta on stainless steel coupons was evaluated, and the effect of cleaning and sanitization procedures in the control of these biofilms was determined. The formation of biofilms was observed while varying the incubation temperature (7, 25 and 39��C) and time (0, 1, 2, 4, 6 and 8days). At 7��C, the counts of E. faecalis and E. faecium were below 2log10CFU/cm(2). For the temperatures of 25 and 39��C, after 1day, the counts of E. faecalis and E. faecium were 5.75 and 6.07log10CFU/cm(2), respectively, which is characteristic of biofilm formation. The tested sanitation procedures a) acid-anionic tensioactive cleaning, b) anionic tensioactive cleaning+sanitizer and c) acid-anionic tensioactive cleaning+sanitizer were effective in removing the biofilms, reducing the counts to levels below 0.4log10CFU/cm(2). The sanitizer biguanide was the least effective, and peracetic acid was the most effective. These studies revealed the ability of enterococci to form biofilms and the importance of the cleaning step and the type of sanitizer used in sanitation processes for the effective removal of biofilms.

Ocorr��ncia de fungos filamentosos termo-resistentes em polpa de tomate envasada assepticamente

This work aimed at determining the occurrence of heat resistant molds during the aseptic processing of tomato pulp (8�� BRIX). During tomato harvest, 9 lots were sampled (3 at the beginning, 3 at the apex and 3 at the end of harvest) and other 5 lots were sampled between harvest. For each lot, the enumeration of heat resistant molds was carried out in samples collected during the aseptic process. The mean count of heat resistant molds was relatively low, ranging from &lt;1 to 8CFU/100mL of sample. The higher counts were observed in the raw material and the pre-wash and transportation water. Fifty strains of heat resistant molds detected in the enumeration procedure were isolated, codified and stocked. One-month-old spores of each isolate were submitted to different heat shocks to select the most heat resistant mold. The most heat resistant isolated strain (survived 100�� C/25 minutes) was identified as Neosartorya fischeri.

Influence of the length of remaining root canal filling and post space preparation on the coronal leakage of Enterococcus faecalis

This study evaluated the sealing ability of different lengths of remaining root canal filling and post space preparation against coronal leakage of Enterococcus faecalis. Forty-one roots of maxillary incisors were biomechanically prepared, maintaining standardized canal diameter at the middle and coronal thirds. The roots were autoclaved and all subsequent steps were undertaken in a laminar flow chamber. The canals of 33 roots were obturated with AH Plus sealer and gutta-percha. The root canal fillings were reduced to 3 predetermined lengths (n=11): G1=6 mm, G2=4 mm and G3=2 mm. The remaining roots served as positive and negative controls. Bacterial leakage test apparatuses were fabricated with the roots attached to Eppendorf tubes keeping 2 mm of apex submerged in BHI in glass flasks. The specimens received an E. faecalis inoculum of 1 x 107 cfu/mL every 3 days and were observed for bacterial leakage daily during 60 days. Data were submitted to ANOVA, Tukey's test and Fisher's test. At 60 days, G1 (6 mm) and G2 (4 mm) presented statistically similar results (p>0.05) (54.4% of specimens with bacterial leakage) and both groups differed significantly (p<0.01) from G3 (2 mm), which presented 100% of specimens with E. faecalis leakage. It may be concluded that the shortest endodontic obturation remnant leaked considerably more than the other lengths, although none of the tested conditions avoids coronal leakage of E. faecalis.

Photodynamic therapy for root canals infected with Enterococcus faecalis

Objective: The aim of this study was to investigate the effects of photodynamic therapy (PDT) on endodontic pathogens by evaluating the decrease in numbers of Enterococcus faecalis colonies in the canals of extracted human teeth. Background Data: Failure in endodontics is usually related to inadequate cleaning and disinfection of the root canal system. This is due to the establishment of microorganisms in areas where the instruments and chemical agents used during root canal preparation cannot eliminate them. PDT is a complementary therapeutic method that could be used to eliminate these remaining bacteria. PDT is a process in which radiation acts on a dye that is applied to the target organism, resulting in bacterial death. Materials and Methods: Forty-six uniradicular teeth had their canals contaminated with bacteria and were incubated for 48 h at 35 degrees C. After that, the teeth were divided into a control group (CG) and a test group (TG). The 23 CG teeth did not undergo any intervention, whereas in the TG the teeth received a solution of 0.0125% toluidine blue for 5 min followed by irradiation using a 50-mW diode laser (Ga-Al-As) at a wavelength of 660 nm. Bacterial samples were taken before and after irradiation. In each of the samples, the number of colony-forming units (CFU) was counted. Results: The mean decrease in CFU was 99.9% in the TG, whereas in the CG an increase of 2.6% was observed. Conclusion: PDT was effective as a bactericidal agent in Enterococcus faecalis-contaminated root canals.