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Biomedical subjects

S A Sattar

Publications and source records attributed to S A Sattar.

10 recordsLinked to original sources

Simian rotavirus SA-11 plaque formation in the presence of trypsin.

Incorporation of 5 micrograms of trpsin per ml of the overlay (Eagle minimal essential medium-0.7% Ionagar no. 2) was found to be necessary for plaque formation by simian rotavirus SA-11. Plaques of 3 to 4 mm in diameter were produced in MA-104 cells after 5 days of incubation at 37 degrees C. Plaque size was even larger (5 to 6 mm) in monolayers of African green monkey kidney cells. Addition of diethyl-aminotheyl-dextran, protamine sulfate, or 5-bromodeoxyuridine to the trypsin-containing overlay did not improve plaque formation by the virus. Incorporation of beef extract or yeast extract to a final concentration of 0.5% in the trypsin-containing overlay inhibited plaque formation. On the other hand, the presence of lactalbumin hydrolysate or peptone at a similar concentration in the overlay did not inhibit plaque formation. When methylcellulose was used instead of the agar as the solidifying agent in the overlay, no plaques were seen. SA-11 is a useful model for the study of human rotaviruses, and this relatively simple plaque assay system should further enhance its usefulness in this regard.

Animals

Second-step concentration of viruses in drinking and surface waters using polyethylene glycol hydroextraction.

In our laboratory, virus adsorbed to talc--Celite layers is eluted with 100 mL of 10% fetal calf serum (FCS) in normal saline (pH 9.0). A further 10-fold reduction in the volume of the eluate was necessary before its inoculation into cell cultures. A 100-mL volume of an experimentally contaminated sample was placed in a dialysis sac and hydroextracted overnight (4 degrees C) with polyethylene glycol (PEG) 6000. The viscous material remaining in the sac was resuspended in 10 mL of Earle's balanced salt solution. After membrane filtration (0.2 micron), the concentrate was plaque assayed in BS-C-1 cells. Using this technique, recoveries of five laboratory-adapted enteric viruses (polio 1, echo 6, coxsackie B5, coxackie A9, and reo 3) and four freshly isolated enteric virus strains (polio 1, echo 1, coxsackie B3, and reo) ranged from 87 to 97%. In comparative tests, PEG hydroextraction was simpler and superior to organic flocculation.

Enterovirus

Recovery of viruses from field samples of raw, digested, and lagoon-dried sludges.

In a 22-month study, viruses were detected in 84% (62/74) of raw, 53% (19/36) of anaerobically digested, and 39% (11/28) of lagoon-dried sludge samples. Lagoon sludge contained detectable viruses (reovirus and enterovirus groups) even after 8 months of retention. Because of such prolonged virus survival in sludge, care must be taken in its disposal or utilization.

Sewage

Viruses in sewage: effect of phosphate removal with calcium hydroxide (lime).

During calcium hydroxide (lime) treatment (pH 9.6 to 10.5) of wastewaters for phosphate removal there was also a two-log removal of added poliovirus (type I, Sabin) from effluents. A similar virus reduction was seen in the sludge generated in these experiments. However, in view of the limitations of techniques for virus recovery from sludge, only a small portion of the infectious virus present in lime sludge may have been detected. Storage of lime sludge at 28 degrees C for up to 48 h produced no appreciable reduction in the virus titre. Five sets of field samples of sewage, effluents, and sludge from a sewage treatment plant (Kemptville, Ont.) which utilizes lime for phosphate removal were also examined for indigenous viruses being BS-C-1 cells. All of the sample of lime sludge and 80% of the samples of both sewage and lime-treated effluent revealed virus; after chlorination only 20% of the lime-treated effluent samples were positive for virus. In contrast, in an earlier study with essentially the same experimental set up, 76% of the sample of chlorinated primary effluent were found to contain virus. Because of the easily detectable quantities of infectious virus in lime sludge and due to the lack of virus inactivation during storage of such sludge, caution must be exercised in its handling and disposal.

Antiviral Agents

Isolation of apparently wild strains of poliovirus type 1 from sewage in the Ottawa area.

In the first 4 months of 1974, 140 gauze pad samples of sewage collected in the Ottawa area were analysed by the BS-C-1 cell system for the presence of viruses pathogenic for humans. Viruses were isolated from 111 (79%) of the samples. Of the 72 (65%) isolates identified by serology and electron microscopic examination, 56 (78%) were reoviruses and 16 (22%), enteroviruses. The enterovirus isolates included one coxsackievirus B4, one vaccine strain of poliovirus type 3, nine vaccine strains of poliovirus type 1 and five strains of poliovirus type 1 that proved by serodifferentiation and temperature marker tests to be different from vaccine strains. The fact that these strains were present in the community sewage in readily detectable concentrations at a time when immunity against polioviruses is declining in such communities is a cause for concern.

Humans

Comparison of four eluents in the recovery of indigenous viruses from raw sludge.

The efficiency of 3% casein hydrolysate (CH), 3% lactalbumin hydrolysate (LH), 3% beef extract (BE), and 10% fetal calf serum (FCS) was compared for the recovery of viruses from raw sludge. CH and LH proved to be inefficient and were eliminated from the study after initial testing. In tests with 20 different samples of raw sludge, beef extract eluted virus in 15 (75%) and FCS revealed virus in 19 (95%) of the samples using BS-C-1 cells. That different eluents were not eluting different viruses from the same sample was shown by the serologic and electron-microscopic examination of 43% (18/42) of the isolates. The identified viruses included members of the entero- (coxsackie B, and polio) and reo-virus groups.

Animals

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.

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 X 10(5) 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.

Adsorption

Lactoglobulins (goat) in the immunofluorescence staining of parainfluenza virus type 2 in L-132 cells.

Instillation of parainfluenza virus type 2 in the mammary gland of the goat resulted in the appearance of specific and high titered antibodies in the milk. The globulin fraction of the milk was separated and conjugated with fluorescein isothiocyanate. These conjugated antibodies could be successfully used in the specific staining of the homologous virus in infected L-132 cells. Therefore, milk from such immunized animals represents a rich and convenient source of specific antibodies for use in immunofluorescence and other serological tests.

Animals