Bacterial Numbers on Penicylinders Used in Disinfectant Testing: Use of 24 Hour Adjusted Broth Cultures

1988 ◽  
Vol 71 (1) ◽  
pp. 9-11
Author(s):  
Eugene C Cole ◽  
William A Rutala

Abstract The current AOAC use-dilution methods of disinfectant efficacy testing require the use of 48-54 h unadjusted broth cultures of Salmonella choleraesuis, Staphylococcus aureus, and Pseudomonas aeruginosa for the inoculation of stainless steel penicylinders. The use of unadjusted broth cultures contributes to noncomparable numbers of organisms on penicylinders among the test strains due to relative efficacy of bacterial attachment to penicylinders and to bacterial numbers in broth. To achieve comparable numbers of cells on the penicylinders among the 3 test strains, the cell densities of S. aureus and P. aeruginosa in broth culture were visually adjusted. Growth studies were conducted using S. choleraesuis and P. aeruginosa to determine the numbers of cells in broth at timed intervals and the corresponding numbers of cells attaching to the penicylinders. Results showed that the use of the 24 h broth cultures for all 3 test strains, with adjustment of S. aureus and P. aeruginosa broths, contributes to more comparable numbers of organisms attached to the penicylinders used in disinfectant testing.

1988 ◽  
Vol 71 (5) ◽  
pp. 868-871
Author(s):  
Edith M Alfano ◽  
Eugene C Cole ◽  
William A Rut Ala

Abstract Stainless steel penicylinders inoculated separately with test bacteria {Salmonella choleraesuis, Pseudomonas aeruginosa, or Staphylococcus aureus) are used in the AOAC use-dilution method (UDM) for disinfectant efficacy testing. Numbers of bacteria remaining on penicylinders were quantitatively assessed to determine if cells are washed from the penicylinders after a 10 min exposure to phosphate buffer dilution water (PBDW). Inoculated penicylinders were also examined by scanning electron microscopy (SEM) to determine the presence of cells remaining attached to the penicylinders after a 10 min exposure to a quaternary ammonium disinfectant and separately to PBDW. The percentage of cells washed from inoculated penicylinders exposed to PBDW was 89.9 for Salmonella choleraesuis, 48.8 for Pseudomonas aeruginosa, and 38.8 for Staphylococcus aureus. Qualitative examination of penicylinders by scanning electron microscopy confirmed the attachment of S. aureus and P. aeruginosa cells to penicylinders exposed separately to PBDW and a quaternary ammonium disinfectant. Few S. choleraesuis cells were observed on penicylinders exposed to PBDW and no cells were observed after disinfectant exposure. The variability of the numbers of viable cells entering the recovery media among the 3 UDM test bacteria due to cell detachment could be a significant factor in the recognized variability of the use-dilution method.


2018 ◽  
Vol 21 (0) ◽  
Author(s):  
Flávia Cíntia de Oliveira ◽  
Tamara Rezende Marques ◽  
Gustavo Henrique Andrade Machado ◽  
Thaís Cristina Lima de Carvalho ◽  
Aline Aparecida Caetano ◽  
...  

Abstract The phenolic compounds from various extracts of jabuticaba skin powder (JSP) were characterized in this study, and the antibacterial activity assessed. The phenolic compounds were extracted from the JSP using four methods: a) acetone extraction - 1 g JSP: 10 mL 70% acetone, resting for 2 hours; b) aqueous extract - 1 g JSP: 15 mL water, under agitation; c) ethanolic extract - 1 g JSP: 15 mL acidified ethanol, under agitation; and d) methanolic extract - 1 g JSP: 50 mL 50% methanol, under reflux. The antibacterial activity was evaluated by the agar diffusion assay, using Escherichia coli ATCC 11229, Salmonella choleraesuis ATCC 6539, Pseudomonas aeruginosa ATCC 15442, Staphylococcus aureus ATCC 6538 and Listeria monocytogenes ATCC 19117. The ethanolic and methanolic extracts showed the highest levels of phenolic compounds, especially of cyanidin chloride, catechin and epicatechin. The extracts did not inhibit the growth of Escherichia coli and Salmonella choleraesuis, but inhibited 30% of the growth of Pseudomonas aeruginosa with an extract concentration of 250 µg mL-1. Against Staphylococcus aureus and Listeria monocytogenes the highest inhibitory effect observed was 41.8% for the ethanolic extract, followed by 36% inhibition by the methanolic extract, thus revealing the potential of these extracts as possible alternatives for use in the food and/or pharmaceutical industries.


Author(s):  
Luz Karime Medina-cÓrdoba ◽  
Ligia Lucia Valencia-mosquera ◽  
Gretty Paola Tarazona-diaz ◽  
Janeth Del Carmen Arias-palacios

Objective: To evaluate the efficacy of a disinfectant based on hydrogen peroxide.Methods: The method used to assess the efficacy of the disinfectant was the agar plate technique. With this procedure, it was possible to determine the percentage of inhibition of the high-level disinfectant of STERIS against four microorganisms, i.e., Pseudomonas aeruginosa ATCC 9027, Staphylococcus aureus (Beta-Hemolytic 227), Salmonella choleraesuis (Kuznedorf CMDM 074), and Bacillus subtilis (ATCC 6633). The effectiveness of five disinfectant concentrations (0.02%, 0.04%, 0.08%, 1%, and 2%) was determined and evaluated in three different times 5, 10, and 15 min, for vegetative strains and 3, 6, and 9 h for the sporulated strain.Results: According to the experimental test, the reduction of the microbial population was, on average, 100% for the disinfectant concentrations of 0.08%, 1%, and 2%.Conclusion: The results obtained demonstrated that the high-level disinfectant of STERIS based on hydrogen peroxide is 100% effective when the concentration recommended by the commercial house (2%) is used in the shortest time exposure to disinfectant. The minimum level of effectiveness was 0.08%; however, if lower concentrations are used, destruction of the microorganisms is not guaranteed.


1987 ◽  
Vol 70 (5) ◽  
pp. 903-906
Author(s):  
Eugene C Cole ◽  
William A Rutala ◽  
Johnny L Carson

Abstract Two passible deficiencies in the AOAC use-dilution method for registration of chemical disinfectants by the Environmental Protection Agency are examined: (7) the physical disparities among brands of penicylinders and (2) the variability of bacterial numbers on penicylinders depending upon test strain and penicylinder surface texture. Textual differences of 2 brands of stainless steel penicylinders, one brand of porcelain, and one brand of glass were assessed by scanning electron microscopy. A considerable variation in smoothness of both inner and outer surfaces of stainless steel and porcelain penicylinders was observed. Glass penicylinders were very smooth. Numbers of bacteria attached to a penicylinder were assessed by vortexing the penicylinders 30 s at No. 4 after using the AOAC method of bacterial inoculation and drying 40 min at 37°C. With this methodology, stainless steel carriers retained the 3 AOAC-recommended bacterial test strains differentially: ca 107 for Pseudomonas aeruginosa, 5 x 106 tat Staphylococcus aureus, and 106 for Salmonella choleraesuis; glass retained 10'-107 organisms of all 3 test strains; porcelain retained about that amount of S. aureus but 105-106 P. aeruginosa and 103-104 S. choleraesuis. These data suggest that disinfectants are not similarly challenged with the AOAC-recommended test bacteria and that an alternative method should be considered to ensure comparable numbers of bacteria on penicylinders


Author(s):  
Neda Fazeli ◽  
Akram Sadat Naeemi ◽  
Seyed Amir Hossein Jalali ◽  
Hojjatollah Zamani

Background: Staphylococcus aureus and Pseudomonas aeruginosa are important human bacterial pathogens, which are resistant to several antibiotics. One of the main causes of their resistance is the ability of biofilm formation. Objectives: The present study aimed to evaluate the antibacterial and antibiofilm activity of the extracts of Vibrio parahaemolyticus, V. alginolyticus, Pseudoalteromonas gelatinilytica, and Pseudoalteromonas piscicida isolated from sea anemone (Stichodactyla haddoni) against S. aureus and P. aeruginosa. Methods: Four isolated bacteria were identified using biochemical and molecular identification methods, and their extracts were obtained by mixing the cell-free supernatants from their old broth culture using ethyl acetate and methanol as the solvents. The agar well-diffusion and micro-dilution methods were also applied to determine the antibacterial activity, minimum bactericidal concentration (MBC), and minimum inhibitory concentration (MIC) of the extracts. The ability of the extracts to inhibit biofilm formation and disrupt the preformed biofilm of the pathogens was attained through crystal violet staining in 96-well microtiter plates. To determine the nature of the extracts, they were exposed to protease enzyme, and the antibiofilm activity was compared with the untreated extracts. Results: The extracts of the four isolated bacteria inhibited bacterial growth and biofilm formation and disrupted the preformed biofilm of S. aureus (MIC = BIC = 600 µg/mL) and P. aeruginosa (MIC = BIC = 300 µg/mL). In addition, the active compounds of the extracts with antibiofilm activities were mainly proteases. Conclusions: According to the results, V. parahaemolyticus, V. alginolyticus, P. gelatinilytica, and P. piscicida had antibacterial and antibiofilm potential against S. aureus and P. aeruginosa, and their extract could also be further analyzed as an alternative to antibiotics.


2021 ◽  
Vol 10 (9) ◽  
pp. e22610918051
Author(s):  
Soraya Castro Trindade ◽  
Jurandi Nery Pereira-Filho ◽  
Laerte Oliveira Barreto Neto ◽  
Thais Brito de Oliveira Moura ◽  
Antonio Pedro Fróes de Farias ◽  
...  

As espécies do gênero Lippia (Verbenaceae) possuem diversas moléculas bioativas, oriundas do seu metabolismo secundário, que apresentam atividade antimicrobiana, analgésica, antifúngica e imunomoduladora. Este estudo teve como objetivo avaliar a atividade antimicrobiana in vitro do extrato metanólico das folhas de L. alnifolia, L. origanoides, L. insignis e L. thymoides frente a microrganismos de importância clínica.  Foi realizada uma triagem fitoquímica dos extratos das plantas e a avaliação da atividade antimicrobiana pelo método de difusão em disco, além da determinação da concentração inibitória mínima (MIC) frente às bactérias Bacillus subtilis, Escherichia coli, Micrococcus luteus, Pseudomonas aeruginosa, Staphylococcus aureus e Salmonella choleraesuis, e os fungos Candida albicans e Candida glabrata. A presença de saponinas, terpenos, esteroides, cumarinas, ácidos fenólicos e flavonoides foi observada na análise fitoquímica. Os extratos metanólicos das folhas de L. alnifolia, L. origanoides e L. insignis apresentaram atividade antimicrobiana contra todas os microrganismos testados. O extrato metanólico da L. thymoides apresentou atividade antimicrobiana frente aos microrganismos M. luteus, P. aeruginosa, S. aureus, C. albicans e C. glabrata. A maioria dos extratos apresentou atividade bacteriostática e todos os extratos apresentaram atividade fungistática para C. albicans e fungicida para C. glabrata. A utilização dos extratos de Lippia pode ser uma alternativa viável no tratamento de doenças infecciosas de origem bacteriana e fúngica.


2009 ◽  
Vol 75 (11) ◽  
pp. 3586-3592 ◽  
Author(s):  
Phat L. Tran ◽  
Adrienne A. Hammond ◽  
Thomas Mosley ◽  
Janette Cortez ◽  
Tracy Gray ◽  
...  

ABSTRACT Among the most difficult bacterial infections encountered in treating patients are wound infections, which may occur in burn victims, patients with traumatic wounds, necrotic lesions in people with diabetes, and patients with surgical wounds. Within a wound, infecting bacteria frequently develop biofilms. Many current wound dressings are impregnated with antimicrobial agents, such as silver or antibiotics. Diffusion of the agent(s) from the dressing may damage or destroy nearby healthy tissue as well as compromise the effectiveness of the dressing. In contrast, the antimicrobial agent selenium can be covalently attached to the surfaces of a dressing, prolonging its effectiveness. We examined the effectiveness of an organoselenium coating on cellulose discs in inhibiting Pseudomonas aeruginosa and Staphylococcus aureus biofilm formation. Colony biofilm assays revealed that cellulose discs coated with organoselenium completely inhibited P. aeruginosa and S. aureus biofilm formation. Scanning electron microscopy of the cellulose discs confirmed these results. Additionally, the coating on the cellulose discs was stable and effective after a week of incubation in phosphate-buffered saline. These results demonstrate that 0.2% selenium in a coating on cellulose discs effectively inhibits bacterial attachment and biofilm formation and that, unlike other antimicrobial agents, longer periods of exposure to an aqueous environment do not compromise the effectiveness of the coating.


1972 ◽  
Vol 55 (1) ◽  
pp. 219-223
Author(s):  
John H S Chen ◽  
L F Ortenzio

Abstract A simple turbidimetric method has been developed for determining the degree of phenol resistance of cultures used for disinfectant testing. Net transmission values are used to assess t h e degree of resistance; these are based on t h e measurement of the total amount of light scattered by the bacterial growth i n the AOAC broth medium without interfering turbidity of the dissolved ingredients. A decrease in the net transmission value corresponds to a decrease in phenol resistance; the converse is also true . The transmission values obtained in a 10 month study of a total of 77 weekly r and om samples of AOAC nutrient broth media blanks showed no substantial differences among the broth media tested by the turbidimetric method. Net transmission values were obtained for 71 broth cultures of Salmonella choleraesuis, 74 of Staphylococcus aureus, and 78 of Pseudomonas aeruginosa having the same required resistance against phenol. These values, which fell within a wider range than would be expected, are given. The effect of the number of 10-carrier-soakings on the net transmission value and equivalent phenol resistance of the cultures was determined for 60 broth cultures of the 3 test organisms named above. Soaking wet cylinder carriers in a given volume of test cultures used for the AOAC use-dilution test directly reduced the net transmission value and equivalently decreased the phenol resistance of the cultures. The turbidimetric method has proven experimentally satisfactory i n several hundred determinations and in selecting test cultures of uniform quality for disinfectant testing.


Author(s):  
Yi Yi Yong ◽  
Michelle Wei Kim Ong ◽  
Gary Dykes ◽  
Wee Sim Choo

Abstract Staphylococcus aureus and Pseudomonas aeruginosa are bacteria that cause biofilm-associated infections. The aim of this study was to determine the activity of combined betacyanin fractions from Amaranthus dubius (red spinach) and Hylocereus polyrhizus (red pitahaya) against biofilms formed by co-culture of S. aureus and P. aeruginosa on different polymer surfaces. Various formulations containing different concentrations of the betacyanin fractions were investigated for biofilm inhibiting activity on polystyrene surfaces using the crystal violet assay and scanning electron microscopy. A combination of each betacyanin fraction (0.625 mg mL−1) reduced biofilm formation of five S. aureus strains and four P. aeruginosa strains from OD values of 1.24–3.84 and 1.25–3.52 to 0.81–2.63 and 0.80–1.71, respectively. These combined fractions also significantly inhibited dual-species biofilms by 2.30 and reduced 1.0–1.3 log CFU cm−2 bacterial attachment on polymer surfaces such as polyvinyl chloride, polyethylene, polypropylene and silicone rubber. This study demonstrated an increase in biofilm inhibiting activity against biofilms formed by two species using combined fractions than that of using single fractions. Betacyanins found in different plants could collectively be used to potentially decrease the risk of biofilm-associated infections caused by these bacteria on hydrophobic polymers.


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