A highly efficient second-step concentration technique for bacteriophages and enteric viruses using ammonium sulfate and Tween 80

1988 ◽  
Vol 34 (5) ◽  
pp. 651-655 ◽  
Author(s):  
Robert Armon ◽  
Max Arella ◽  
Pierre Payment
Keyword(s):  
Ammonium Sulfate ◽  
Tween 80 ◽  
Protective Properties ◽  
Concentration Technique ◽  
Potassium Tartrate ◽  
Coxsackievirus B ◽  
Virus Recovery ◽  
Nonionic Detergents ◽  
Floating Layer

Addition of Tween 80 to a 1.5% solution of beef extract was found to enhance the elution of bacteriophages adsorbed to electronegative filters. When reconcentration of the eluate was attempted by ammonium sulfate precipitation, a floating layer containing most of the viruses was formed. This floating layer can be obtained with several nonionic detergents including Tween 80 and under a salt saturation of 55% with ammonium sulfate, potassium tartrate, and sodium phosphate. Virus recovery ranged from 91 to 103% and was obtained with several bacteriophage strains. With poliovirus type 1, coxsackievirus B-4, and rotavirus SA-11 the recoveries were 100, 20, and 80%, respectively, but toxicity to cell culture was encountered: after removal of the detergent by a second floating layer method the recovery was 32% for poliovirus. Compared with organic flocculation, this method also had both improved recovery for bacteriophages and protective properties for samples frozen at −70 °C.

Second-step reconcentration of environmental samples by ammonium sulfate flocculation of beef extract

1987 ◽  
Vol 33 (6) ◽  
pp. 571-572 ◽  
Author(s):  
Pierre Payment ◽  
Michel Trudel
Keyword(s):  
Ammonium Sulfate ◽  
Environmental Samples ◽  
Enteric Viruses ◽  
Second Step ◽  
Poliovirus Type ◽  
Beef Extract ◽  
Coxsackievirus B ◽  
Acid Environment

Some enteric viruses are sensitive to the acid environment utilized during the concentration of viruses from water. The use of a neutral flocculant, neutralized ammonium sulfate at 50% saturation, permitted the recovery of 97% of the simian rotavirus SA-11, 87% of Coxsackievirus B-4, and 88% of poliovirus type 1. This method should permit a better recovery of enteric viruses from the environment.

scholarly journals Developing a vaccine for type 1 diabetes by targeting coxsackievirus B

2018 ◽  
Vol 17 (12) ◽  
pp. 1071-1083 ◽  
Author(s):  
Heikki Hyöty ◽  
Francisco Leon ◽  
Mikael Knip
Keyword(s):  
Type 1 Diabetes ◽  
Coxsackievirus B

Modified Procedure for Extraction of Poliovirus from Naturally-Infected Oysters Using Cat-Floc and Beef Extract

1980 ◽  
Vol 43 (2) ◽  
pp. 91-94 ◽  
Author(s):  
EDWARD F. LANDRY ◽  
JAMES M. VAUGHN ◽  
THOMAS J. VICALE
Keyword(s):  
Crassostrea Gigas ◽  
Extraction Procedure ◽  
Acid Precipitation ◽  
Poliovirus Type ◽  
Extraction Procedures ◽  
Precipitation Technique ◽  
Beef Extract ◽  
Virus Recovery

Methods for recovery of poliovirus type 1 (LSc2ab) from naturally-infected oysters (Crassostrea gigas) were examined. Extraction procedures analyzed included glycine-saline and polyelectrolyte (Cat-Floc) methods followed by concentration using modifications of an acid precipitation technique, Direct viral assay of shellfish homogenates, when compared to virus recovery following extraction, indicated that substantially fewer viruses were detected in initial homogenates. These data appeared to support the contention that input values based on homogenate assay were inappropriate in determining recovery efficiencies with naturally-infected shellfish. Since absolute efficiencies could not be determined, relative efficiencies using samples from pooled homogenates were used to determine the recovery efficiencies of various extraction procedures. Cat-Floc extraction followed by a beef extract-modified acid precipitation technique resulted in higher virus recoveries than a glycine-saline extraction procedure.

scholarly journals CHARACTERIZATION OF CELLULASE PREPARATION OF BACILLUS SP.G4 ISOLATED FROM THE TERMITES GUT

2018 ◽  
Vol 54 (4A) ◽  
pp. 115
Author(s):  
Đào Thị Thanh Xuân
Keyword(s):  
High Activity ◽  
Ammonium Sulfate ◽  
Rice Bran ◽  
Tween 80 ◽  
Bacillus Sp ◽  
High Concentration ◽  
Acetone Precipitation ◽  
Cmcase Activity ◽  
Purification Factor

The Bacillus sp. strain G4 isolated from termites gut can produce cellulase with high activity in rice bran medium after 72 h of incubation at 37 °C. The crude enzymewas collected by centrifugation at 6,000 rpmfor 15 minutes.  The precipitation of cellulasewith ammonium sulfate 60 – 90 % saturation and acetone at the concentration 60 % – 90 % was studied. Results showed that ammonium sulfate 90 % saturation gave cellulase with highest purification factor 8.87 but the yield was only 59.9 %, whereas acetone at 90 % concentration gave highest yield 83.57 % with purification factor 4.38. CMCase activity of cellulasepreparation obtained by acetone precipitation at 90 % was optimum at pH 7 and 60 °C. Furthermore, CMCase was stable at pH 6.0 - 7.0 and at temperature lower than 50 °C. The CMCase was activated by ion Ca2+ but inhibited by Co2+, Zn2+, Cu2+, Fe2+, and was not affected by Mg2+ and Ba2+. The CMCase was inhibited by high concentration of SDS while EDTA and Tween 80 played activated role.

Comparison of talc-Celite and polyelectrolyte 60 in virus recovery from sewage: development of technique and experiments with poliovirus (type 1, Sabin)-contaminated multilitre samples

1976 ◽  
Vol 22 (11) ◽  
pp. 1620-1627 ◽  
Author(s):  
Syed A. Sattar ◽  
J. C. N. Westwood
Keyword(s):  
Fetal Calf Serum ◽  
Calf Serum ◽  
Sample Pretreatment ◽  
Sewage Sample ◽  
Type I ◽  
Poliovirus Type ◽  
Ph Adjustment ◽  
Field Samples ◽  
Virus Recovery

For virus recovery from sewage, a mixture of talc and Celite was tested as a possible inexpensive substitute for polyelectrolyte 60(PE 60).After adjustment of pH to 6 and the addition of 45–60 plaque forming units (PFU)/ml of poliovirus type I(Sabin) to the sewage sample under test, 100 ml of it was passed through either a PE 60 (400 mg) or a talc (300 mg)–Celite (100 mg) layer; the layer-adsorbed virus was eluted with 10 ml of 10% fetal calf serum (FCS) in saline (pH 7.2).In these experiments, PE 60 layers recovered 73–80% (mean 76%) of the input virus. In comparison, virus recoveries with the talc–Celite layers were 65–70% (mean 68%).Passage of 5 litres of raw sewage (containing 50 to 1.26 × 105 PFU/100 ml of the poliovirus) through the talc (15 g)–Celite (5 g) layers and virus elution with 50 ml of 10% FCS in saline gave virus recoveries of 33–63% (mean 49%).Except for pH adjustment and prefiltration through two layers of gauze to remove large solids, no other sample pretreatment was found to be necessary. Application of this technique to recovery of indigenous viruses from field samples of raw sewage and effluents has been highly satisfactory.

scholarly journals A hexavalent Coxsackievirus B vaccine is highly immunogenic and has a strong protective capacity in mice and nonhuman primates

2020 ◽  
Vol 6 (19) ◽  
pp. eaaz2433 ◽  
Author(s):  
V. M. Stone ◽  
M. M. Hankaniemi ◽  
O. H. Laitinen ◽  
A. B. Sioofy-Khojine ◽  
A. Lin ◽  
...  
Keyword(s):  
Nonhuman Primates ◽  
Neutralizing Antibody ◽  
Proof Of Concept ◽  
Murine Models ◽  
Human Pathogens ◽  
Protective Capacity ◽  
Coxsackievirus B ◽  
Excellent Safety Profile ◽  
High Immunogenicity

Coxsackievirus B (CVB) enteroviruses are common human pathogens known to cause severe diseases including myocarditis, chronic dilated cardiomyopathy, and aseptic meningitis. CVBs are also hypothesized to be a causal factor in type 1 diabetes. Vaccines against CVBs are not currently available, and here we describe the generation and preclinical testing of a novel hexavalent vaccine targeting the six known CVB serotypes. We show that the vaccine has an excellent safety profile in murine models and nonhuman primates and that it induces strong neutralizing antibody responses to the six serotypes in both species without an adjuvant. We also demonstrate that the vaccine provides immunity against acute CVB infections in mice, including CVB infections known to cause virus-induced myocarditis. In addition, it blocks CVB-induced diabetes in a genetically permissive mouse model. Our preclinical proof-of-concept studies demonstrate the successful generation of a promising hexavalent CVB vaccine with high immunogenicity capable of preventing CVB-induced diseases.

Test Method for Determination of Virucidal Efficacy of Liquid Surface Disinfectants

1976 ◽  
Vol 59 (5) ◽  
pp. 1150-1155
Author(s):  
Charles R McDuff ◽  
John W Gaustad
Keyword(s):  
Collaborative Study ◽  
Test Procedure ◽  
Test Method ◽  
Virus Concentration ◽  
Ammonium Compound ◽  
Concentration Technique ◽  
Modified Method ◽  
Culture Cells

Abstract A test method was developed incorporating AOAC use-dilution stainless steel Penicylinders and H.Ep.-2 tissue culture cells for the evaluation of disinfectants as virucides. The subsequent collaborative study using Herpes simplex and poliovirus type 1 in evaluation against a quaternary ammonium compound, phenolic, and iodiphor disinfectants indicated that the basic test procedure is sound. However, an improved virus concentration technique and a modified method for drying virus-contaminated cylinders are recommended. In addition, other types of disinfectants and test viruses should be included in order to demonstrate that the procedure is versatile as well as reproducible.

scholarly journals A Coxsackievirus B vaccine protects against virus-induced diabetes in an experimental mouse model of type 1 diabetes

Diabetologia ◽  
2017 ◽  
Vol 61 (2) ◽  
pp. 476-481 ◽  
Author(s):  
Virginia M. Stone ◽  
Minna M. Hankaniemi ◽  
Emma Svedin ◽  
Amirbabak Sioofy-Khojine ◽  
Sami Oikarinen ◽  
...  
Keyword(s):  
Type 1 Diabetes ◽  
Mouse Model ◽  
Coxsackievirus B ◽  
Experimental Mouse Model ◽  
Experimental Mouse
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