Bacillus cereus e Geobacillus stearothermophilus em leite ultra alta temperatura (UAT) / Bacillus cereus and Geobacillus steartothermophilusin ultra high temperature (UHT) milk

O objetivo deste trabalho foi avaliar a presença de B. cereus e G. stearothermophilus em 100 amostras de leite UAT (integral e desnatado). O isolamento dos esporos e das células vegetativas seguiu metodologias oficiais, com pequenas modificações. B. cereus foi isolada de 7% amostras de leite UHT, de 6 diferentes marcas. As contagens máximas de células vegetativas e esporos de B. cer eus foram de 3,54 Log UFC/mL e 3,93 Log esporos/mL, respectivamente. A presença dos genes codificadores de enterotoxina não hemolítica (NHE) foi observada em 33% dos isolados e da hemolisina (HBL ) em 100% dos isolados. O gene hblA foi encontrado em 91,6 % dos isolados, porém nenhum isolado apresentou os 3 genes do complexo HBL. G. stearothermophilus foi identificada em 22,8% (34/149) dos isolados de esporo altamente resistente ao calor (HRRS), representando 18% das amostras de leite UAT e as contagens de esporos variaram de < 1Log a 3,40 Log esporos/mL.

Enzymatic saccharification and fermentation of cellulosic date palm wastes to glucose and lactic acid

Abstract The bioconversion of cellulosic wastes into high-value bio-products by saccharification and fermentation processes is an important step that can reduce the environmental pollution caused by agricultural wastes. In this study, enzymatic saccharification of treated and untreated date palm cellulosic wastes by the cellulases from Geobacillus stearothermophilus was optimized. The alkaline pre-treatment of the date palm wastes was found to be effective in increasing the saccharification percentage. The maximum rate of saccharification was found at a substrate concentration of 4% and enzyme concentration of 30 FPU/g of substrate. The optimum pH and temperature for the bioconversions were 5.0 and 50 °C, respectively, after 24 h of incubation, with a yield of 31.56 mg/mL of glucose at a saccharification degree of 71.03%. The saccharification was increased to 94.88% by removal of the hydrolysate after 24 h by using a two-step hydrolysis. Significant lactic acid production (27.8 mg/mL) was obtained by separate saccharification and fermentation after 72 h of incubation. The results indicate that production of fermentable sugar and lactic acid is feasible and may reduce environmental pollution by using date palm wastes as a cheap substrate.

Metronomic chemotherapy in the neoadjuvant setting: results of two parallel feasibility trials (TraQme and TAME) in patients with HER2+ and HER2− locally advanced breast cancer

Neoadjuvant chemotherapy has practical and theoretical advantages over adjuvant chemotherapy strategy in breast cancer (BC) management. Moreover, metronomic delivery has a more favorable toxicity profile. The present study examined the feasibility of neoadjuvant metronomic chemotherapy in two cohorts [HER2+ (TraQme) and HER2− (TAME)] of locally advanced BC. Twenty patients were prospectively enrolled (TraQme, n=9; TAME, n=11). Both cohorts received weekly paclitaxel at 100 mg/m2 during 8 weeks followed by weekly doxorubicin at 24 mg/m2 for 9 weeks in combination with oral cyclophosphamide at 100 mg/day (fixed dose). The HER2+ cohort received weekly trastuzumab. The study was interrupted because of safety issues. Thirty-six percent of patients in the TAME cohort and all patients from the TraQme cohort had stage III BC. Of note, 33% from the TraQme cohort and 66% from the TAME cohort displayed hormone receptor positivity in tumor tissue. The pathological complete response rates were 55% and 18% among patients enrolled in the TraQme and TAME cohorts, respectively. Patients in the TraQme cohort had more advanced BC stages at diagnosis, higher-grade pathological classification, and more tumors lacking hormone receptor expression, compared to the TAME cohort. The toxicity profile was also different. Two patients in the TraQme cohort developed pneumonitis, and in the TAME cohort we observed more hematological toxicity and hand-foot syndrome. The neoadjuvant metronomic chemotherapy regimen evaluated in this trial was highly effective in achieving a tumor response, especially in the HER2+ cohort. Pneumonitis was a serious, unexpected adverse event observed in this group. Further larger and randomized trials are warranted to evaluate the association between metronomic chemotherapy and trastuzumab treatment.

Thermostable, alkaline and detergent-tolerant lipase from a newly isolated thermophilic Bacillus stearothermophilus.

Lipases are the enzymes of choice for laundry detergent industries, owing to their triglyceride removing ability from the soiled fabric, which eventually reduces the usage of phosphate-based chemical cleansers in the detergent formulation. In this study, a novel thermo-alkaline lipase-producing strain identified as Bacillus stearothermophilus was isolated from the soil samples of olive oil mill. Enhanced lipase production was observed at 55°C, pH 11 and after 48 h of incubation. Among the substrates tested, xylose (a carbon source), peptone (a nitrogen source) and olive oil at a concentration of 1% were suitable substrates for enhancing lipase production. MgSO4 and Tween-80 were suitable substrates for maximizing lipase production. The enzyme was purified to homogeneity by a single CM-Sephadex column chromatography and revealed molecular mass of 67 kDa. The enzyme (BL1) was active over a wide range of pH from 9.0 to 13.0, with an optimum at pH 11.0, exhibited maximal activity at 55°C and retained more than 70% of its activity after incubation at 70°C or pH 13 for 0.5 h or 24 h, respectively. The enzyme hydrolyzed both short and long-chain triacylglycerols at comparable rates. BL1 was studied in a preliminary evaluation for use in detergent formulation solutions. This novel lipase showed extreme stability towards non-ionic and anionic surfactants after pre-incubation for 1 h at 40°C, and good stability towards oxidizing agents. Additionally, the enzyme showed excellent stability and compatibility with various commercial detergents, suggesting its potential as an additive in detergent formulations.

A study on the incidence, microbiological analysis and investigations on the source of infection of postoperative infectious endophthalmitis in a tertiary care ophthalmic hospital: An 8-year study.

Background: The objective of the study was the determination of the incidence of culture-proven postoperative endophthalmitis and probable sources of infection. Materials and Methods: It was a prospective study on the microbiology, incidence and probable sources of infection in patients with postoperative infectious endophthalmitis carried out in a tertiary care eye hospital. Consecutive patients diagnosed with postoperative infectious endophthalmitis during the years 2000-2007 were investigated for the causative infective agent and possible sources of infection. The surgical data and microbiological data including the investigations performed to trace the source were recorded in a specific formatted form and were gathered and compiled for analysis. Results: Data of analysis showed that 98 (0.042%) out of 2,31,259 patients who underwent intra-ocular surgery developed infectious endophthalmitis. Among these, 70 (0.053%) occurred after cataract, 10 (0.5%) after penetrating keratoplasty (PK) and 18 (0.018%) following other types of intra-ocular surgeries. The predominant infectious agents isolated were bacteria (89.7%), with equal proportions of Gram-positive and Gram-negative bacteria. Polymicrobial infection was noted in four and fungi in seven patients. Occurrence of postoperative endophthalmitis was sporadic and not related to any specific part of period in a year. Sources of infection were donor corneal rim in six post-PK patients and phaco probe in one who had postphacoemulsification endophthalmitis Conclusions: Overall incidence of postoperative endophthalmitis over an 8-year period was quite low. The sources of infection could be established in six post-PK endophthalmitis patients and in a postcataract surgery.

Esterilização em Ortodontia: eficácia do esterilizador com esferas de vidro / Sterilization in Orthodontics: effectiveness of glass bead sterilizers

OBJETIVO: avaliar a efetividade do esterilizador com esferas de vidro Steri® 350, quanto ao controle de infecção das partes ativas dos alicates ortodônticos. METODOLOGIA: foram utilizados nove alicates ortodônticos, previamente esterilizados em autoclave à temperatura de 121ºC, durante 20 minutos. Posteriormente, as partes ativas dos alicates foram contaminadas com Bacillus stearothermophilus. Logo após, as pontas dos alicates foram colocadas no Steri® 350, durante os períodos de 3, 5, 10, 15, 20, 30 e 40 segundos, em temperatura de 255ºC, para avaliar a eficácia da esterilização. RESULTADOS E CONCLUSÕES: o esterilizador com esferas de vidro mostrou-se eficaz no controle do crescimento de Bacillus stearothermophilus nas partes ativas dos alicates ortodônticos, a partir de 10 segundos de exposição à temperatura de 255ºC.

AIM: The aim of this research was to evaluate the efficiency of Steri® 350glass bead sterilizer, on the infection control of the active part of the orthodontic pliers. METHODS: Nine orthodontic pliers were sterilized in the steam autoclave at 121ºC in 20 minutes and subsequently the active part of each plier was dipped in a culture for Bacillus stearothermophilus. Later, the active part of each plier was putted into the Steri® 350 in the following periods: 3, 5, 10, 15, 20, 30, and 40 seconds at 255ºC to evaluate the efficiency of this method in the infection control. RESULTS AND CONCLUSIONS: This sterilization method has been effective to control the presence of Bacillus stearothermophilus on the active part of orthodontic pliers at 255ºC in 10 seconds.

Screening of phytate hydrolysing marine bacteria isolated from Baluchistan

A total of 100 Marine bacterial strains were isolated and screened for phytate-degrading ability. The phytate degrading ability of the isolates was first qualitatively evaluated by the formation of haloes [clear zones] around the colonies growing on solid medium containing 10mM of sodium phytate and then quantitatively evaluated by measuring amount of free phosphate released as a result of sodium phytate hydrolysis by bacterial strains using high performance liquid chromatography and by spectrophotometric method. Thirteen percent of the isolated strains showed sodium phytate-degrading ability in solid medium and three percent of the strains showed sodium phytate degrading ability in liquid medium. The strains, which showed hydrolysis of sodium phytate both in solid and liquid media, were identified as E. coli, Bacillus steriothermophilus, and Pseudomonas aeruginosa. E.coli released 3.5mM phosphate and Bacillus steriothermophilus released 2.4mM free phosphate where as Pseudomonas aeruginosa released 1.4mM free phosphate

Geobacillus stearothermophilus LV cadA gene mediates resistance to cadmium, lead and zinc in zntA mutants of Salmonella entérica serovar Typhimurium

Salmonella entérica serovar Typhimurium cells expressing the cadA gene of Geobacillus stearothermophilus LV exhibit a hypersensitive phenotype to cadmium chloride. Deletion of the ORF STM3576 from the Salmonella genome resulted in cadmium, lead and zinc sensitivity, confirming that this ORF is a homologue of the zntA gene. The observed sensitivity was reverted upon expression of the G. stearothermophilus LV cadA gene. These results indicate that the cadA gene product is involved in Cd, Pb and Zn resistance as a classical P-type ATPase and strongly suggest that the observed hypersensitive phenotype to these metals can be related to the function of the host ·zntA gene product.

Antimicrobial activity of two brazilian commercial propolis extracts

The antimicrobial activity of propolis extracts is well documented, but little is known about the antimicrobial properties of commercial products containing propolis, since these vary according to the geographical region in which the propolis is obtained. This study evaluated the antimicrobial activity of two samples of commercial propolis on 26 species of microorganisms obtained from ATCC and some wild strains: Gram-positive cocci and bacilli, and Gram-negative rods and yeasts. The tested products were two samples of Brazilian commercial propolis from Apis Flora™: 11.0% ethanolic extract of propolis (EEP) and Propomax™ 11.0% extract of propolis without alcohol (EP). Antimicrobial activity was determined by the agar diffusion technique, well method. MIC was determined for Staphylococcus sp. and Streptococcus mutans using the method of broth dilution with the propolis extract in serial concentrations. EEP and EP showed antimicrobial activity against all tested bacteria and yeasts, having a more pronounced action against Gram-positive bacteria and Candida albicans ATCC 10231, and a less evident activity against Gram-negative and Candida albicans FT2010. For S. mutans, the EEP MIC ranged from 8.8 to 4.4 mL of propolis, and the EP MIC, from 4.4 to <1.1 mL. For Staphylococcus sp., the MIC of both extracts was <1.1.

In Vivo and In Vitro evaluation of the efficacy of a peracetic acid-based disinfectant for decontamination of acrylic resins

O objetivo deste trabalho foi avaliar a eficácia antimicrobiológica de um desinfetante à base de ácido peracético na descontaminação de resinas acrílicas termicamente ativada, quimicamente ativada e polimerizada em forno de microondas. Placas de resina foram contaminadas in vivo por meio do uso intraoral por 10 voluntários durante 7 noites e corpos-de-prova de resina foram contaminados in vitro por meio do contato com microrganismos conhecidos: Bacillus subtilis e Bacillus stearothermophilus. Os espécimes contaminados foram imersos em desinfetante à base de ácido peracético a 0,2% (Sterilife®; Lifemed) durante 5 ou 10 min e então colocados no meio de cultura BHI. Após incubação a 37°C for 48 h, o crescimento bacteriano foi avaliado por meio análise da turvação do meio de cultura. Todos os espécimes imersos em ácido peracético por 5 ou 10 min não apresentaram turvação do meio de cultura, enquanto os espécimes contaminados e colocados diretamente no meio de cultura (grupo controle) apresentaram turvação. Concluiu-se que a imersão em ácido peracético por pelo menos 5 min foi eficaz na desinfecção de resinas acrílicas termicamente ativada, quimicamente ativada e polimerizada em forno de microondas contaminadas tanto com saliva humana quanto com Bacillus subtilis ou Bacillus stearothermophilus.

Evaluation of a Bacillus stearothermophilus tube test as a screening tool for anticoccidial residues in poultry

A Bacillus stearothermophilus var. calidolactis C953 tube test was evaluated for its ability in detecting the residue of selected anticoccidial drugs in poultry, specically sulfamethazine, furazolidone, and amprolium. Various concentrations of each drug were injected into chicken liver and kidney tissues and these tissues were tested to determine the drug detection limits for each drug. The detection limit was defined as the drug concentration at which 95% of the test results were interpreted as positive. The limits of detection in liver tissue were 0.35 microgram/ml for furazolidone, 0.70 microgram/ml for sulfamethazine and 7.80 microgram/ ml for amprolium. In kidney tissues, they were 0.30 microgram/ml for furazolidone, 0.54 microgram/ml for sulfamethazine, and 7.6 microgram/ml for amprolium. It was concluded that this tube test could be used to screen for the residue of these three drugs in poultry.

Viable spore counts in biological controls pre-sterilization

El objetivo de este trabajo fue evaluar el recuento total de esporos viables en tiras inoculadas estandarizadas antes de ser sometidas a ciclo de esterilización. Se utilizaron muestras de "Bacterial Spore Sterilization Strip" (R. Biological Lab) dentro de la fecha de vencimiento, que fueron divididas en: grupo A (B subtilis) y en grupo B (B stearothermophylus). Se testearon 24 tiras de cada grupo. Las tiras, en grupos de tres, fueros trituradas y colocadas en agua destilada refrigerada estéril. Luego se homogeneizaron las suspensiones en vortex durante 5 minutos y se transfirieron 10 ml de cada grupo a dos frascos estériles. Las muestras se calentaron en baño de maría a 95ºC (grupo A y 80ºC (grupo B) durante 15 minutos y se las enfrió bruscamente en un baño de hielo entre 0º a 4ºC durante 15 minutos. Se realizaron diluciones sucesivas hasta obtener una alícuta final que estuviera entre 30 y 300 UFC (unidades formadoras de colonias) que fue colocada en placas de Petri que contenían el medio de cultivo (agar extracto de soja caseína fundido, adaptado para esporos y enfriado a 45-50ºC. Se realizaron las incubaciones a 55ºC y 37ºC. El número de esporos presentes a las 48 horas fue mayor que el número presente a las 24 horas, si bien estos resultados no fueron homogéneos en todos los grupos. Los datos fueron analizados estadísticamente. El controlar el recuento de esporos viables pre-esterilización evitaría falsos resultados. La cantidad de esporos viables no debe ser menor al 50 por ciento ni mayor al 300 por ciento del número de esporos de la cepa específica en el control biológico. Este procedimiento es importante para garantizar la eficacia del control biológico

Procesamiento a distancia de controles biológicos: parte I / Distance processing of biological controls: part I

Los controles biológicos son indicadores de eficacia del proceso de esterilización y avalan la destrucción de toda forma de vida microbiana y sus formas de resistencia. El objetivo de este estudio fue evaluar controles biológicos en ciclos de esterilizacion. Grupo I (n:64) método aleatorio de control biológico (tierra con esporos de Bacillus sp. y Clostridium sp.), Grupo II (n:64), portador inoculado con esporos B. stearothermophilus 1.6 x 10 4 esporoso y B. subtilis var. niger 2.3 x 10 6 esporos por tira) y Grupo III (n:64) alambres de acero inoxidable. La totalidad de las muestras (n:192) fueron acondicionadas dentro de envoltorios de papel de uso médico conteniendo cada una, por duplicado, los grupos I, II y III. La mitad (n:96) se esterilizó por estufa (E) y las restantes (n:96) por autoclave (A). Posteriormente se sembraron las muestras en los siguientes tiempos: inmediato a la esterilización, a los 30, 60 y 100 días y se evaluaron los resultados. En los grupos experimentales no se encontraron diferencias estadísticamente significativas por la prueba del Chi2 (p=0,09) entre los grupos I y II en los tiempos analizados. Los resultados sugieren que ambos controles son efectivos y el procesamiento puede extenderse hasta los tres meses posesterilización.

Validação do processamento térmico de um produto protéico vegetal enlatado / Thermal processing validation of a canned vegetal protein product

A validação de processos nas indústrias alimentícia e farmacêutica é uma das principais ferramentas da garantia de qualidade, tornando os produtos seguros, eficazes e confiáveis. Realizou-se a validação física e biológica da esterilização terminal de um produto protéico constituído da proteína texturizada de soja, proteína de trigo (glúten), gordura vegetal hidrogenada e condimentos, envasado em latas metálicas de 850 gramas. A validação física do autoclave foi executada em três ciclos na câmara com 12 sensores de temperatura. Confirmou-se para os pontos geometricamente distribuídos no interior do autoclave que a diferença máxima de temperatura entre um ponto e outro foi de 1,0°C em relação à temperatura média da câmara (121°C)...

Overexpression and characterization of dimeric and tetrameric forms of recombinant serine hydroxymethyltransferase from Bacillus stearothermophilus.

Serine hydroxymethyltransferase (SHMT), a pyridoxal-5' -phosphate (PLP) dependent enzyme catalyzes the interconversion of L-Ser and Gly using tetrahydrofolate as a substrate. The gene encoding for SHMT was amplified by PCR from genomic DNA of Bacillus stearothermophilus and the PCR product was cloned and overexpressed in Escherichia coli. The purified recombinant enzyme was isolated as a mixture of dimer (90%) and tetramer (10%). This is the first report demonstrating the existence of SHMT as a dimer and tetramer in the same organism. The specific activities at 37 C of the dimeric and tetrameric forms were 6 7 U/mg and 4 1 U/mg, respectively. The purified dimer was extremely thermostable with a T(m) of 85 degrees C in the presence of PLP and L-Ser. The temperature optimum of the dimer was 80 degrees C with a specific activity of 32 4 U/mg at this temperature. The enzyme catalyzed tetrahydrofolate-independent reactions at a slower rate compared to the tetrahydrofolate-dependent retro-aldol cleavage of L-Ser. The interaction with substrates and their analogues indicated that the orientation of PLP ring of B. stearothermophilus SHMT was probably different from sheep liver cytosolic recombinant SHMT (scSHMT).

Estudo sobre a expressão dos genes de alfa amilase de Bacillus Stearothermophilus e de Anospheles merus em células de bactéria, levedura e inseto / Studies on the gene expression of Bacillus stearothermophilus and Anopheles merus alpha amylases in bacteria, yeast, insect cells

O gene A1 da alfa amilase foi isolado de uma biblioteca genômica em lambda EMBL3, feita com DNA do díptero primitivo Anopheles merus (Díptera, Nematocera, Culicoidea). Ele foi parcialmente seqüênciado, caracterizado pelo padrão da clivagem das enzimas restritivas e clonado no plasmídeo pIBI24 por Pernasetti (1991). No presente trabalho relata-se sua expressão em: bactéria (Eschirichia coli AD494), células de inseto (Spodoptera frugiperda) e em levedura (Pichia pastoris), tendo-se observado que somente nas células de Spodoptera frugiperda a enzima é expressa com atividade. A expressão extracelular do gene A1 foi ensaiada em E. coli AD494 usando o vetor pRSETc...

Endosporos termófilos del ambiente azucarero de Jujuy / Thermophilic endospores in the environment of a sugar mill in Jujuy

Twenty six samples from green and scorched sugarcane stems and leaves, sugarmill air dust and raw sugar were analyzed. Thirty nine thermophilic bacilli strains were isolated. Physiological and morphological examinations were carried out according to Bergey's Manual. The strains were identified as B. licheniformis (66.7), B. coagulans (17.9), B. stearothermophilus (10.3) y B. subtilis (5.1).

Verificación biológica de los ciclos de esterilización / Biologic verification of the sterilization cycles

En 55 meses se realizaron 2,920 pruebas con IB en 91 consultorios dentales de la República Mexicana. El 71.4 por ciento (n=2,084) de los ciclos de esterilización fue en vapor de agua a presión, el 9.4 por ciento (n=274) vapor químico a presión, y el 19.2 por ciento (n=562) en calor seco. El 67.6 por ciento (n=1974) de las pruebas se realizó en forma semanal, el 16.9 por ciento (n=493) quincenal y el 15.5 por ciento (n=453) mensual. Se detectaron fallas en 7.6 por ciento (n=223) de todos los ciclos de esterilización. El 7.5 por ciento (n=156) en vapor de agua a presión, el 7.7 por ciento (n=21) en vapor químico a presión y el 8.2 por ciento (n=46) de los ciclos de calor seco. Todos los métodos de esterilización empleados fallaron con frecuencias similares (X = 0.307, P mayor 0.8)

Análisis de algunos indicadores biológicos disponibles comercialmente en México / Analysis of some biological markers available in Mexico

El objetivo de esta investigación fue comparar, mediante las pruebas descriptas en la farmacopea de los Estados Unidos Mexicanos, diversos indicadores biológicos (IB) contra las cepas de Bacillus subtilis 9372 y B. stearothermophilus 7953 del catálogo de la ATCC. Se observó que las cepas ATCC y los IB provenientes de fabricantes extranjeros cumplen con las características morfológicas, bioquímicas y de cultivo estipuladas por la FEUM. De hecho, los fabricantes extranjeros establecen que sus productos están elaborados con las cepas ATCC. Por el contrario, algunos IB de fabricación nacional no son elaborados con las cepas ATCC. El aislado de B. subtilis, para verificar la esterilización por calor seco, se comporta como la cepa ATCC-9372, mientras que los indicadores para vapor a presión, supuestamente B. stearothermophilus, se comportan como B. subtilis sin pigmento. El fabricante de IB deberá documentar la veracidad de sus afirmaciones y proporcionar información que permita al usuario la selección de productos adecuados. Las instrucciones de uso deberán ser precisas