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PubMed1981

Urea-lysine method for recovery of enteroviruses from sludge.

Enteroviruses added to sludge and indigenous viruses present in sludge were recovered by treating the sludge flocs with a 4 M urea solution buffered at pH 9 with 0.5 M lysine. Eluted viruses were absorbed to aluminum hydroxide flocs and collected by centrifugation. The flocs were solubilized with 0.1 M ethylenediaminetetraacetic acid-3% beef extract at pH 9. After dialysis to remove the ethylenediaminetraacetic acid, viruses were further concentrated by organic flocculation. Approximately 40% of poliovirus and coxsackievirus B-3 added to 500 to 1,000 ml of sludge could be recovered in final sample volumes of less than 10 ml. Polioviruses, echoviruses, and coxsackieviruses were recovered from different samples of wastewater sludge.

Centrifugation↗

PubMed1982

Comparison of methods for recovering indigenous viruses from raw wastewater sludge.

Five general methods for recovering indigenous viruses from raw wastewater sludge were compared. Each method included elution, concentration, and disinfection steps. The elution method, found to consistently yield the greatest viral recovery, was a two-phase technique that involved blending sludge with Freon. Other methods, including two being tested as American Society for Testing Materials tentative standard methods, were less effective. Viral recoveries were generally greater (sometimes much greater) if samples were concentrated by high-speed centrifugation rather than by organic flocculation with 3% beef extract. Three cell lines were used to measure viral recoveries by the plaque assay. The efficiency of recovery was greatest on BGM cells, followed by RD and MA-104 cells.

Cell Line↗

PubMed1982

Recovery of viruses from vegetable surfaces.

The efficiency of a system developed for the recovery of viruses contaminating large quantities of vegetables was investigated in the laboratory and tested in the field. Viruses seeded onto a number of leafy vegetables in the laboratory were eluted with a phosphate-buffered saline solution (pH 9.0). The eluate was clarified by glass wool filtration, and any viruses present were concentrated by adsorption to a Filterite pleated cartridge filter, eluted with 3% beef extract (pH 9.0), and further concentrated by organic flocculation. At least 24 liters of vegetable eluate could be concentrated to 70 to 80 ml, equivalent to a greater than 99.5% reduction in volume. With this system, poliovirus was recovered with a mean efficiency of 58% for all vegetables tested. Adenovirus was recovered from lettuce with a slightly lower mean efficiency (55%). Poliovirus was recovered from large quantities of cabbage for up to 5 days in the field after spray irrigation of relatively low levels of virus, even when heavy rain fell before sampling.

Adenoviridae↗

PubMed1983

Method for recovery of enteric viruses from estuarine sediments with chaotropic agents.

An evaluation was made of the ability of chaotropes, low-molecular-weight ionic compounds which enhance the solubilization of hydrophobic compounds in water, to improve the recovery of enteric viruses from highly organic estuarine sediments. Chaotropic agents alone were poor eluents of polioviruses from sediment but were effective when combined with 3% beef extract. Chaotropes of lower potency, NaNO3, NaCl, and KCl, were more efficient eluents than the stronger chaotropes, guanidium hydrochloride or sodium trichloroacetate. The most effective eluent was 2 M NaNO3 in 3% beef extract at pH 5.5, which eluted 71% of sediment-associated polioviruses. Efficient concentration of the sodium nitrate-beef extract eluate by organic flocculation required the addition of the antichaotrope (NH4)2SO4 to a 2 M concentration and Cat-Floc T (Calgon, Pittsburgh, Pa.) a cationic polyelectrolyte, to a 0.01% concentration. Dialysis of the final concentrate was necessary to reduce salts to nontoxic levels before assay in cell cultures. Trials with highly organic estuarine sediment seeded with high or low numbers of poliovirus 1, echovirus 1, or rotavirus SA-11 demonstrated the superiority of this method over two other methods currently in use.

Enterovirus↗

PubMed1987

Methods for recovering poliovirus and rotavirus from oysters.

Polioviruses and rotaviruses are potential indicators of sewage pollution of water and shellfish. Several methods for detecting these viruses in oysters were assessed. Elution-precipitation involving Catfloc for clarification and skim milk for subsequent flocculation resulted in the recovery of an average of 79% of poliovirus type 1 and 37% of rotavirus SA-11 from oyster homogenates inoculated with low numbers of these viruses. Adsorption-elution-precipitation did not improve the recovery of poliovirus and was detrimental to the recovery of rotavirus. Ultrafiltration or ultracentrifugation resulted in improved recovery of rotavirus but also in higher toxicity of oyster extracts to cell cultures. We recommend the use of the described elution-precipitation method for detecting viral pollutants in sample of oysters.

Animals↗

PubMed1962

Tetanus toxin production in the absence of protein.

A modification of the Mueller medium for tetanus toxin production is presented, on which an adapted strain produces high yields of toxin (70 to 90 flocculating units per ml) in the absence of animal protein extracts. The range of iron concentration in the medium was established within which the toxigenic activity of the culture can be made to vary, and the mathematical nature of the variation is presented. Darkening of cultures during incubation indicates departure from optimal conditions. Heat input to the medium during autoclaving, originally undertaken solely for purposes of sterilization, is an important physicochemical factor in the toxigenicity of the culture.

Animals↗

PubMed1972

Identification of two Saccharomyces cerevisiae cell wall mannan chemotypes.

We have obtained evidence for two structurally and antigenically different Saccharomyces cerevisiae cell wall mannans. One, which occurs widely and is found in S. cerevisiae strain 238C, is already known to be a neutral mannan which yields mannose, mannobiose, mannotriose, and mannotetraose on acetolysis of the (1 --> 6)-linked backbone. The other, which was found in S. cerevisiae brewer's strains, is a phosphomannan with a structure very similar to that of Kloeckera brevis mannan. S. cerevisiae (brewer's yeast strain) was agglutinated by antiserum prepared against Kloeckera brevis cells. The mannan, isolated from a proteolytic digest of the cell wall of the former, did not react with S. cerevisiae 238C antiserum, whereas it cross-reacted strongly with K. brevis antiserum. Controlled acetolysis cleaved the (1 --> 6)-linkages in the polysaccharide backbone and released mannose, mannobiose, mannotriose, and mannotriose phosphate. Mild acid treatment of the phosphomannan hydrolyzed the phosphodiester linkage, yielding phosphomonoester mannan and mannose. The resulting phosphomonoester mannan reacted with antiserum prepared against K. brevis possessing monoester phosphate groups on the cell surface. alpha-d-Mannose-1-phosphate completely inhibited the precipitin reaction between brewer's yeast mannan and the homologous antiserum. Flocculent and nonflocculent strains of this yeast were shown to have similar structural and immunological properties.

Agglutination Tests↗

PubMed1983

Presumptive identification of Vibrio species with H antiserum.

Species in the genus Vibrio exhibit flagellar (H) antigens unique to the species. Thus, species-specific H antiserum could be a valuable reagent with which to screen serologically large numbers of Vibrio isolates. Antisera against V. cholerae, V. fluvialis, V. anguillarum, V. metschnikovii, V. parahaemolyticus, V. alginolyticus, and V. vulnificus H antigens was produced in rabbits by repeated injections of Formalin-preserved whole cells. Anti-O activity and anti-H activity against common H antigens was absorbed from each antiserum, V. fluvialis was shown to possess an H antigen unique to the species and also to share minor H antigens with V. cholerae, V. metschnikovii, and V. anguillarum. V. vulnificus also exhibits a species-unique H antigen. A comprehensive serological screening system based on species-specific H antiserum was developed to identify pathogenic Vibrio species one step beyond primary isolation. Vibrio species were correctly identified with accuracies ranging from 93 to 100%. Some isolates were either nonmotile or poorly so and thus did not flocculate in H antiserum.

Antibodies, Viral↗

PubMed1975

Comparison of the counter-immunoelectrophoresis technique with the Reiter protein and three other serological tests as a first line test for syphilis.

The counter-immunoelectrophoresis technique with the Reiter protein (RP-CIE) was compared with two complement fixation tests (Kolmer and RPCF) and a flocculation test (VDRL) in sensitivity and specificity. Of the 1,927 consecutive attendants of a venereal disease clinic whose serum samples were used, 250 were considered to be syphilitic. The number of true-positive and false-positive reactions were: 121 and 4 (VDRL), 124 and 2 (Kolmer), 179 and 41 (RPCF), 166 and 16 (RP-CIE). The VDRL and the RPCF combined were more sensitive and less specific than the VDRL combined with the Kolmer. If the RPCF was replaced by the RP-CIE the sensitivity remained the same but the specificity was higher. The RP-CIE scored more positives than the RPCF in untreated cases of primary syphilis. The results argue for substitution of the RP-CIE for the Kolmer in the combination with the VDRL in the serodiagnosis for syphilis. Moreover, the RP-CIE presents the technical advantages of simplicity, speed of performance, and of not being hampered by the anticomplementary nature of the serum sample.

Antigens, Bacterial↗

PubMed1977

The effect of hyperimmunization with Neisseria gonorrhoeae on the presence of gonococcal antibody in serum, tissues, and secretions of the rabbit.

Antibody responses in sera, tissues, and secretions of the urogenital tract and lower respiratory tract of rabbits hyperimmunized with Neisseria gonorrhoeae were examined. Antibody was detected by passive hemagglutination, whole-cell agglutination, bentonite flocculation, and in some cases immunodiffusion-in-gel. Immunization of rabbits either intravenously or intramuscularly resulted in the presence of gonococcal antibodies in the sera, spleens, and tissue of the urogenital tract (vagina, cervix, uterus, and fallopian tubes). Gonococcal antibody was also found in secretions bathing the mucosa of the urogenital tract and lower respiratory tract. Antibodies were not detected in sera, tissues, and secretions of non-immunized rabbits. The spleen was shown to synthesize gonococcal antibody in vitro in response to hyperimmunization. Tissues of the urogenital tract did not appear to synthesize gonococcal antibody thus suggesting and antibodies present in secretions of the urogenital tract were derived mainly from serum.

Agglutination Tests↗

PubMed1979

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↗

PubMed2005

Influence of carbon source and surface hydrophobicity on the aggregation of the yeast Kluyveromyces bulgaricus.

Aggregation of the yeast Kluyveromyces bulgaricus is mediated by the galactose-specific lectin KbCWL1. This lectin contains hydrophobic amino acids and its activity is calcium dependent. A specific fluorescent probe, 1-anilinonaphthalene-8-sulfonic acid in the free acid form (ANS; Sigma Chemical Co., St. Louis, Missouri), was used to study the hydrophobic areas on the cellular surface of K. bulgaricus. Changes in surface hydrophobicity during the growth and aggregation of yeast cells were studied. Surface hydrophobicity increased during growth and depended on the amount of yeast cells in the culture medium. During growth, the size of the hydrophobic areas on the cell surface was measured using ANS and was found to increase with the percentage of flocculating yeasts. Our results strongly suggest that the hydrophobic areas of the cell walls of yeast cells are involved in the aggregation of K. bulgaricus.

Anilino Naphthalenesulfonates↗

PubMed1975

The effect of barium on blood in the gastrointestinal tract.

The effect of various commercial barium sulfate preparations on blood coagulation, flocculation, and hemolysis was studied. There was considerable variation not only among different barium products but also among different concentrations of the same product. There is no good experimental evidence that barium enemas do stop bleeding. It is probably best to avoid the use of any enema in attempting to control major colonic bleeding; but if an enema is given for this purpose, a tap-water enema is better, as it contains no anticoagulants and is more effective in producing clot formation than most barium suspensions.

Barium Sulfate↗

PubMed1977

Observations on the behaviour of barium sulphate suspensions in gastric secretion.

A number of barium sulphate suspensions showed varying patterns of stability when provoked with gastric secretion. The stability of the suspensions was affected by the pH, the mucin content and the volume of secretion used. These studies indicate that flocculation of the suspension in the presence of gastric residue decreases as the amount of undiluted barium sulphate in the mixture is increased.

Adsorption↗

PubMed1994

Production of a bioflocculant by mixed culture.

Colony groups that form large amounts of slime externally were obtained from activated sludge among phthalate-assimilating microbes. That slime, which R-3 mixed microbes produced externally, had a high level of flocculation activity. R-3 mixed strains, one of such colony groups, efficiently produced a bioflocculant (APR-3) in liquid cultures of production medium, especially that containing starch and glucose (1:1) as carbon sources. Identification of this group of bioflocculant-producing microbes (R-3 mixed strains) showed that it was comprised of four strains belonging to the genera Oerskovia, Acinetobacter, Agrobacterium, and Enterobacter. The bioflocculant (APR-3) was purified electrophoretically to homogeneity by ethanol and CPC precipitation. Its molecular mass is at least 2 x 10(6) Da. APR-3 is an acidic polysaccharide made of glucose, galactose, succinic acid, and pyruvic acid (molar ratio: 5.6:1:0.6:2.5).

Acinetobacter↗

PubMed2003

Riverine inputs of organic carbon and nitrogen to Norwegian coastal areas.

The transport of organic and inorganic material in 10 Norwegian rivers has been monitored from 1990 to 1998. The suspended material in the rivers consisted of 40-80% organic materials. A major part of this was DOC with a C/ N mole ratio > 25. The C/N ratio of the organic material increases with increasing river flow when drained from forest-dominated catchments, in contrast to mountain dominated catchments where variation in runoff does not influence the C/N ratio. Laboratory experiments showed that more than 90% of the DOC remained in solution when salinity increased to 30. The flocculated and settled material contained less than 5% organic material with a C/N ratio of 6.7-12.4, comparable to C/N ratios in cultivated soils and coastal marine sediments. This confirms that only a minor amount of organic material carried to the sea by rivers settles in the coastal zone.

Carbon↗

PubMedDate not supplied

The influence of vegetation on sedimentation and resuspension of soil particles in small constructed wetlands.

When initiatives to mitigate soil erosion are insufficient or fail, constructed surface flow wetlands (CWs) could be a final buffer to reduce pollution before reaching recipients. The objective of this study was to determine the influence of CW vegetation on the retention of soil particles from arable land. Retention was measured with water flow-proportional sampling systems in the inlet and outlet, sedimentation traps, and sedimentation plates in four small CWs over a period of 5 yr. The surface area of the CWs was 265 to 900 m2, and the average hydraulic loads were 1.2 to 3.4 m d(-1). Watershed areas were 0.5 to 1.5 km2. Annual soil particle retention was 30 to 80% or 14 to 121 kg m(-2). Results show that macrophytes stimulate sediment retention by mitigating resuspension of CW sediment. Five years after construction, resuspension had decreased approximately 40% and was negligible. As vegetation cover increases, the influence of macrophytes on soil particle retention reaches a level where other factors, such as hydraulic load and sediment load, were more important. Macrophytes increased the hydraulic efficiency by reducing short-circuit or preferential flow. However, vegetation did not have any influence on the clay concentration in the sediment. Hence, a possible stimulation of particle flocculation was not detected. Vegetation makes it possible to use the positive effect of a short particle settling distance in shallow ponds by hindering resuspension.

Conservation of Natural Resources↗

PubMedDate not supplied

Particulate and dissolved phosphorus chemical separation and phosphorus release from treated dairy manure.

In confined animal feeding operations, liquid manure systems present special handling and storage challenges because of the large volume of diluted wastes. Water treatment polymers and mineral phosphorus (P) immobilizing chemicals [AI2(SO4)3 x 18H2O, FeCl3-6H2O, and Class C fly ash] were used to determine particulate and dissolved reactive phosphorus (DRP) reduction mechanisms in high total suspended solid (TSS) dairy manure and the P release from treated manure and amended soils. Co-application exceeded the aggregation level achieved with individual manure amendments and resulted in 80 and 90% reduction in metal salt and polymer rates, respectively. At marginally effective polymer rates between 0.01 and 0.25 g L(-1), maximal aggregation was attained in combination with 1 and 10 g L(-1) of aluminum sulfate (3 and 30 mmol Al3+ L(-1)) and iron chloride (3.7 and 37 mmol Fe3+ L(-1)) in 30 g L(-1) (TSS30) and 100 g L(-1) TSS (TSS100) suspensions, respectively. Fly ash induced particulate destabilization at rates > or = 50 g L(-1) and reduced solution-phase DRP at all rates > or = 1 g L(-1) by 52 and 71% in TSS30 and TSS100 suspensions, respectively. Aluminum and Fe salts also lowered DRP at rates < or = 10 g L(-1) and higher concentrations redispersed particulates and increased DRP due to increased suspension acidity and electrical conductivity. The DRP release from treated manure solids and a Typic Paleudult amended with treated manure was reduced, although the amendments increased Mehlich 3-extractable P. Therefore, the synergism of flocculant types allowed input reduction in aggregation aid chemicals, enhancing particulate and dissolved P separation and immobilization in high TSS liquid manure.

Animal Husbandry↗