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

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.

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Comparison of Three Methods for the Concentration of Poliovirus from Moroccan Shellfish

Journal of Food Protection ◽

10.4315/0362-028x-57.11.996 ◽

1994 ◽

Vol 57 (11) ◽

pp. 996-1000 ◽

Cited By ~ 1

Author(s):

NOURREDINE BOUCHRITI ◽

SAGAR M. GOYAL ◽

ABDELHAK El MARRAKCHI ◽

MOHAMMED JELLAL

Keyword(s):

Mytilus Edulis ◽

Crassostrea Gigas ◽

Acid Precipitation ◽

Precipitation Method ◽

Ruditapes Decussatus ◽

Beef Extract ◽

Dry Milk ◽

Virus Recovery

Three methods were evaluated for the concentration of poliovirus from artificially contaminated oysters (Crassostrea gigas), mussels (Mytilus edulis) and carpet-shell clams (Ruditapes decussatus) grown in Morocco. The methods tested were: an adsorption-elution-precipitation method, a beef extract elution acid-precipitation method, and a non-fat dry milk elution acid-precipitation method. For all shellfish species tested, the adsorption-elution-precipitation method yielded the lowest average virus recovery (27%), whereas the two elution-precipitation methods yielded average virus recoveries of 42% each. The beef extract elution acid-precipitation method yielded the highest virus recovery with clams (53%), whereas non-fat dry milk elution acid-precipitation was advantageous for mussels providing average virus recovery of 47%. For oysters, none of the tested methods gave satisfactory virus recovery. These results point towards the need for the development of better method(s) for the concentration of viruses from Moroccan oysters, while for mussels and clams, the elution-acid precipitation methods may be satisfactory.

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Development of a Method for Recovery of Rotavirus from the Surface of Vegetables

Journal of Food Protection ◽

10.4315/0362-028x-48.3.261 ◽

1985 ◽

Vol 48 (3) ◽

pp. 261-264 ◽

Cited By ~ 10

Author(s):

AMIN S. BADAWY ◽

CHARLES P. GERBA ◽

LEE M. KELLEY

Keyword(s):

Disease Outbreaks ◽

Leafy Vegetables ◽

Contaminated Water ◽

Waterborne Disease ◽

Poliovirus Type ◽

Beef Extract ◽

Human Rotavirus ◽

Virus Recovery

Rotaviruses cause waterborne disease outbreaks of gastroenteritis from sewage contaminated water, but methods have not been available to assess the potential for transmission of rotavirus by uncooked foods. A method was developed for recovery of rotavirus from the surface of vegetables. The simian rotavirus SA-11, used as a model for the human rotavirus, was adsorbed onto lettuce and the effects of various eluents tested for its recovery. The maximum recovery of rotavirus occurred with a solution of 3% beef extract at pH 8.0 after 5 min of exposure. Longer exposure times did not enhance virus recovery. Recovery of rotavirus and poliovirus type 1 (LSC) averaged 80 and 65%, respectively. Recovery of rotavirus from non-leafy vegetables was lower, averaging 44% for celery, carrot and radish. This method should prove useful for assessing the occurrence and survival of rotavirus on uncooked foods.

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Processing Pork Samples for Virus Detection: A Preliminary Study Of Methods

Journal of Food Protection ◽

10.4315/0362-028x-54.1.28 ◽

1991 ◽

Vol 54 (1) ◽

pp. 28-31 ◽

Cited By ~ 1

Author(s):

MING YI DENG

Keyword(s):

Extraction Procedure ◽

Virus Detection ◽

Two Phase ◽

Poliovirus Type ◽

The Mean ◽

Preliminary Study ◽

Virus Recovery ◽

Pork Samples ◽

Model Virus

Extraction and concentration procedures for recovering a model virus from pork samples were investigated. A 20-g sample, experimentally contaminated with attenuated poliovirus type 1, was suspended in 100 ml of glycine-NaOH buffer at pH 8.8 and homogenized in an ice bath. The pork solids in the homogenate were flocculated by centrifugation and filtration. Each of the sample extracts was concentrated by either hydroextraction (HE), ultrafiltration (UF), or the polymer two-phase separation (PTPS), and assayed for viruses by the plaque technique in monolayers of HeLa cells. The experiments indicated that the extraction procedure was effective, and that the mean virus recovery from the samples when concentrated by the HE, the UF, and the PTPS was 53.0, 68.0, and 65.1%, respectively. No plaque-forming units were detected in concentrates prepared from uninoculated pork samples. The extraction and concentration procedures developed in the present study are expected to be applicable to the processing of samples for detecting viruses in pork and merit further work.

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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

Canadian Journal of Microbiology ◽

10.1139/m76-238 ◽

1976 ◽

Vol 22 (11) ◽

pp. 1620-1627 ◽

Cited By ~ 12

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.

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Evaluation de l'efficacité de la technique d'adsorption-élution du poliovirus 1 sur filtres en fibres de verre: application à l'analyse virologique de 100 ml à 1000 ℓ d'eau

Canadian Journal of Microbiology ◽

10.1139/m78-225 ◽

1978 ◽

Vol 24 (11) ◽

pp. 1413-1416 ◽

Cited By ~ 5

Author(s):

Pierre Payment ◽

Michel Trudel ◽

Vytautas Pavilanis

Keyword(s):

Sewage Water ◽

Aluminium Chloride ◽

Elution Volume ◽

Fiber Glass ◽

Poliovirus Type ◽

Beef Extract ◽

Different Types ◽

Overall Efficiency

The efficiency of the adsorption-elution technique using fiber glass filters to concentrate viruses from water was evaluated to detect poliovirus type 1 in drinking, river, and sewage water. At pH 3.5 and with 5 × 10−4 M aluminium chloride more than 99% were adsorbed to a 0.25-μm filter. Beef extract (3%), pH 9, eluted 85–95% of the adsorbed viruses and organic flocculation at pH 3.5 permitted to reconcentrate the viruses in 1/20of the elution volume with a 50–72% efficiency. The overall efficiency of the technique for 100 ml to 1000 ℓ of the different types of water using 102 to 106 PFU was 38 to 58%.

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Recovery of Poliovirus Type 1 from Experimentally Contaminated Shellfish: Evaluation of Different Methods

Water Science & Technology ◽

10.2166/wst.1993.0319 ◽

1993 ◽

Vol 27 (3-4) ◽

pp. 45-48 ◽

Cited By ~ 5

Author(s):

L. Croci ◽

D. De Medici ◽

M. Divizia ◽

R. Gabrieli ◽

L. Toti ◽

...

Keyword(s):

Statistical Analysis ◽

Natural Environment ◽

Artificial Seawater ◽

Viral Gastroenteritis ◽

Poliovirus Type ◽

Tangential Flow ◽

Virus Recovery ◽

Concentration Methods ◽

Elution Step

Outbreaks of viral gastroenteritis caused by ingestion of contaminated shellfish are well documented. Several methods have been described for virus recovery from shellfish, but often they are complicated and time-consuming. In this study we have compared several viral extraction-concentration methods from mussels experimentally contaminated in conditions close to those met in natural environment. Mvtilus galloprovincialis were contaminated by 2 h immersion in artificial seawater seeded with Poliovirus type 1. After the contamination step, the mussel bodies with intervalve water were homogenized by Ultraturax blender using four different elution buffers: PBS pH9.5; 0.05M glycine pH9.5; 0.3% and 3% beef extract pH9.5. The ratio between eluants and homogenates, depending on the used method, was 1:2 or 1:10. After the elution step, virus was concentrated comparing the following techniques: precipitation at acid pH, precipitation with PEG6000, ultracentrifugation, ultrafiltration with Amicon apparatus and tangential flow Minitan system. Statistical analysis was performed using ANOVA program and the results were evaluated.

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Survie de virus entériques adsorbés sur microfibre de verre au cours d'un transport postal

Canadian Journal of Microbiology ◽

10.1139/m81-037 ◽

1981 ◽

Vol 27 (2) ◽

pp. 246-248 ◽

Cited By ~ 2

Author(s):

J. -C. Joret ◽

J. -C. Block

Keyword(s):

Control System ◽

Enteric Viruses ◽

Qualitative Studies ◽

Waste Waters ◽

Solution Ph ◽

Poliovirus Type ◽

Extract Solution ◽

Beef Extract

The survival of enteric viruses (poliovirus type 1, Mahoney strain, and indigenous viruses of waste waters) has been studied after adsorption of the viruses (pH 3.5) on glass microfiber filters.After postal transport, the presence of the viruses was checked on the filters being soaked in a 3% beef extract solution (pH 7.5) either frozen or without protection against heat. Viruses were recovered at a rate of 59 to 65%.For qualitative studies, postal shipment of samples adsorbed on fiberglass may allow extension of a control system for enteroviruses in water. [Journal translation]

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

Canadian Journal of Microbiology ◽

10.1139/m87-098 ◽

1987 ◽

Vol 33 (6) ◽

pp. 571-572 ◽

Cited By ~ 5

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.

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