Control of Listeria monocytogenes growth in soft cheeses by bacteriophage P100

The purpose of this study was to determine the effect of bacteriophage P100 on strains of Listeria monocytogenes in artificially inoculated soft cheeses. A mix of L. monocytogenes 1/2a and Scott A was inoculated in Minas Frescal and Coalho cheeses (approximately 10(5) cfu/g) with the bacteriophage added thereafter (8.3 x 10(7) PFU/g). Samples were analyzed immediately, and then stored at 10 ºC for seven days. At time zero, 30 min post-infection, the bacteriophage P100 reduced L. monocytogenes counts by 2.3 log units in Minas Frescal cheese and by 2.1 log units in Coalho cheese, compared to controls without bacteriophage. However, in samples stored under refrigeration for seven days, the bacteriophage P100 was only weakly antilisterial, with the lowest decimal reduction (DR) for the cheeses: 1.0 log unit for Minas Frescal and 0.8 log units for Coalho cheese. The treatment produced a statistically significant decrease in the counts of viable cells (p < 0.05) and in all assays performed, we observed an increase of approximately one log cycle in the number of viable cells of L. monocytogenes in the samples under refrigeration for seven days. Moreover, a smaller effect of phages was observed. These results, along with other published data, indicate that the effectiveness of the phage treatment depends on the initial concentration of L. monocytogenes, and that a high concentration of phages per unit area is required to ensure sustained inactivation of target pathogens on food surfaces.

Isolation of bacteriophages to multi-drug resistant Enterococci obtained from diabetic foot: A novel antimicrobial agent waiting in the shelf.

Introduction: While foot infections in persons with diabetes are initially treated empirically, therapy directed at known causative organisms may improve the outcome. Many studies have reported on the bacteriology of diabetic foot infections (DFIs), but the results have varied and have often been contradictory. The purpose of the research work is to call attention to a frightening twist in the antibiotic-resistant Enterococci problem in diabetic foot that has not received adequate attention from the medical fraternity and also the pharmaceutical pipeline for new antibiotics is drying up. Materials and Methods: Adult diabetic patients admitted for lower extremity infections from July 2008 to December 2009 in the medical wards and intensive care unit of medical teaching hospitals were included in the study. The extent of the lower extremity infection on admission was assessed based on Wagner's classification from grades I to V. Specimens were collected from the lesions upon admission prior to the initiation of antibiotic therapy or within the first 48 h of admission. Results: During the 18-month prospective study, 32 strains of Enterococcus spp. (26 Enterococcus faecalis and 06 E. faecium) were recovered. Antibiotic sensitivity testing was done by Kirby-Bauer's disk diffusion method. Isolates were screened for high-level aminoglycoside resistance (HLAR). A total of 65.6% of Enterococcus species showed HLAR. Multidrug resistance and concomitant resistance of HLAR strains to other antibiotics were quite high. None of the Enterococcus species was resistant to vancomycin. Conclusion: Multidrug-resistant Enterococci are a real problem and continuous surveillance is necessary. Today, resistance has rendered most of the original antibiotics obsolete for many infections, mandating the development of alternative anti-infection modalities. One of such alternatives stemming up from an old idea is the bacteriophage therapy. In the present study, we could able to demonstrate the viable phages against MDR E. faecalis.

Estudo da proteína de choque térmico GRP788 para o desenvolvimento de um sistema de receptor-ligante para o câncer de próstata / The use of the heat-shock protein GRP78 for the development of a receptor-ligant ssystem in prostate cancer

Terapêuticas mais eficazes são necessárias para o câncer de próstata avançado. A Proteína regulada pela glicose-78 (GRP78) foi recentemente descrita como sendo um possível marcador molecular para a doença. Neste estudo, avaliou-se a GRP78 no desenvolvimento de um sistema de receptor-ligante para o câncer de próstata. Inicialmente, foram criados dois fagos com afinidade à GRP78 e foi demonstrado que ambos se ligam especificamente à proteína in vitro e em células de câncer de próstata. Em seguida, mostrou-se que ambos os fagos com afinidade à GRP78 rastrearam xeno-tumores prostáticos in vivo, e finalmente mostrou-se que os fagos ligam-se especificamente à GRP78 em metástases ósseas de câncer de próstata humano /Improved therapies are needed for advanced prostate cancer. The Glucose-regulated protein-78 (GRP78) was recently described as molecular marker for prostate cancer. We hipothesized that GRP78 could be used in the development of a receptor-ligand system. We initially cloned two GRP78-targeting plasmids (WIFPWIQL and WDLAWMFRLPVG) into a fUSE5-based phage. We showed that both phage bound specifically to GRP78 in vitro and to intact prostate cancer cells, and was internalized by those cells. GRP78-binding phage showed a strong homing in vivo to human prostate cancer xenografts. Finally, we showed that both phage bound specifically to GRP78 expressed in human prostate cancer bone metastases...

Estabilidad de fagos de Lactococcus lactis frente al hipoclorito de sodio y durante el almacenamiento / Stability of Lactococcus lactis phages treated with sodium hypochlorite and during storage

Survival of lytic bacteriophages active against Lactococcus lactis ssp. lactis and ssp. cremoris was determined after treatment with sodium hypochlorite and during storage at 4 degrees C. Three phages were isolated from dairy plants in Argentina (ARG) and the other phages were isolated in the United States of America (US). All of them represent phages that infected cheese manufacture industries and belong to different morphological or serological groups. These phages showed higher survival in M17 broth, buffered with sodium glycerophosphate, than in trypteine soy broth (TSB). Phage populations did not decrease significantly during 14 weeks in M17 broth, whereas in TSB the titers of phage suspensions began to decline around 9 days. In addition, the effect of sodium hypochlorite was more marked in broth than in milk. A higher surviving fraction was obtained in milk, even when tenfold higher concentrations of chlorine were used. The effect of hypochlorite on phages of the same serological group was quite similar and independent of phage morphology. However, phage 137-1, which belongs to other serological group, showed lower resistance to sodium hypochlorite. Comparing the hypochlorite inactivation for ARG and US phages, it was observed that they have their own inactivation values, independently of their origin and morphological group. Long periods of time and high concentrations of chlorine were necessary to reduce the surviving fraction in milk. This indicates that hypochlorite concentrations and times of contact can be critical for the efficiency of the operative sanitization processes.

Electron microscopic study of phages and aberrant structures produced by induction of prophage kappa in Vibrio cholerae el tor cells.

Prophage kappa in V. cholerae el tor strain SLH22(J) could be induced spontaneously or by treatment with nitrofurantoin, though the efficiency of induction was very low (not more than 0.8%). V. cholerae el tor cells were found to release many different aberrant structures of the temperate phage, kappa. These aberrant structures were characterized by density gradient centrifugation and electron microscopy.