RESUMEN
Laboratory investigations were conducted to study the growth dynamics of Pseudomonas pickettii in commercial 0.9% sodium chloride solution under various environmental conditions and to determine the retention of these organisms after challenge through a 0.2-micron cartridge filter system. Low numbers of P. pickettii (1 to 10 CFU/ml of test solution) inoculated into commercial vials containing 5 ml of 0.9% sodium chloride solution and 500-ml volumes of 0.9% sodium chloride solution were shown to proliferate over a 168-h incubation period. These organisms demonstrated growth over a wide range of temperatures (15 to 42 degrees C) in this salt solution, and survival studies at 50, 55, and 60 degrees C indicated that this strain was not unusually resistant to heat (with the times required at a given temperature to reduce the surviving microbial population 10-fold [D-values] being 26.0, 1.9, and 0.7 min, respectively). A challenge test demonstrated that P. pickettii organisms were not completely retained by a 0.2-micron cartridge filter. The number of organisms detected increased from 1 CFU/liter of effluent at 1 to 2 min to a maximum of 176 CFU/liter at 4 to 5 min. Our results indicate that P. pickettii can penetrate a 0.2-micron filtration system and that the passage of organisms and subsequent microbial growth in the filter effluent probably are the mechanisms by which these organisms were recovered from "sterile" commercial 0.9% sodium chloride solution.
Asunto(s)
Contaminación de Medicamentos , Pseudomonas/crecimiento & desarrollo , Terapia Respiratoria , Cloruro de Sodio , Calor , Humanos , Microscopía Electrónica , Pseudomonas/ultraestructura , SolucionesRESUMEN
Non-A, non-B hepatitis was transmitted to eight chimpanzees by intravenous inoculation with antihemophilic materials (factor VIII) that had been implicated in transmission of the disease, with acute-phase liver homogenate from an infected chimpanzee, with acute-phase from an infected chimpanzee, or with chronic-phase plasma from infected chimpanzees. All eight animals developed elevated alanine aminotransferase activity, and all demonstrated unique hepatocyte cytoplasmic tubules at some time during the acute phase of disease. The temporal patterns for tubule appearance in hepatocyte cytoplasm, however, were highly variable, even between chimpanzees given similar inocula. Convoluted membranous structures were found occasionally in early or pre-acute-phase liver biopsy specimens. Aggregates of microtubules were also found in hepatocyte cytoplasm in some acute-phase biopsy specimens. No disease-specific nuclear changes or structures, including clusters or crystalline arrays of virus-like particles, were found in any of the serial liver biopsy specimens from chimpanzees with experimental non-A, non-B hepatitis.
Asunto(s)
Hepatitis C/patología , Hepatitis Viral Humana/patología , Hígado/ultraestructura , Alanina Transaminasa/sangre , Animales , Núcleo Celular/ultraestructura , Citoplasma/ultraestructura , Hepatitis C/sangre , Microscopía Electrónica , Microtúbulos/ultraestructura , Pan troglodytes , Factores de TiempoRESUMEN
Inoculation of eight chimpanzees with factor VIII, factor IX, or "H" strain plasma resulted in enzymatic and histopathologic evidence of non-A/non-B hepatitis in all eight animals. Challenge of two chimpanzees convalescent from factor VIII-induced disease with either factor IX or "H" strain plasma resulted in non-A/non-B hepatitis only in the animal inoculated with factor IX materials. Reciprocal cross-challenge of a chimpanzee convalescent from factor IX-induced disease with factor VIII also produced unequivocal enzymatic and histopathologic evidence of non-A/non-B hepatitis. Cross-challenge of a chimpanzee convalescent from "H" strain-induced non-A/non-B hepatitis with factor VII did not cause a second bout of non-A/non-B hepatitis. These findings suggest the factor VIII materials and "H" strain plasma used in these studies share a common etiologic agent (or agents), but that factor VIII and factor IX may contain two distinct agents. Electron microscopic (EM) examination of thin-sectioned, acute-phase liver biopsies from all but one of the chimpanzees receiving the primary inocula revealed the presence of abnormal hepatocyte cytoplasmic structures previously shown to be associated with non-A/non-B hepatitis. Crystalline structure containing 25 to 30 nm particles were visualized by EM in the cytoplasm of endothelial or Kupffer cells in acute-phase liver biopsies obtained from three chimpanzees inoculated with either factor VIII materials or "H" strain plasma.
Asunto(s)
Hepatitis C/inmunología , Hepatitis Viral Humana/inmunología , Inmunidad , Hígado/ultraestructura , Animales , Factor IX/administración & dosificación , Factor IX/efectos adversos , Factor VIII/administración & dosificación , Factor VIII/efectos adversos , Hepatitis C/etiología , Hepatitis C/patología , Pan troglodytes , Reacción a la TransfusiónRESUMEN
Heavy density HAV was also shown to be sensitive to low concentrations of RNase. The results of these biophysical and biochemical studies strongly support the notion HAV is an enterovirus.
Asunto(s)
Brotes de Enfermedades , Hepatitis A/microbiología , Hepatovirus , Antígenos Virales/análisis , Centrifugación por Gradiente de Densidad , Desoxirribonucleasas/metabolismo , Heces/microbiología , Hepatovirus/análisis , Hepatovirus/clasificación , Hepatovirus/inmunología , Humanos , Concentración de Iones de Hidrógeno , Virus ARN/clasificación , Ribonucleasas/metabolismoRESUMEN
Hepatitis A virus (HAV) recovered from stools of human cases of hepatitis A and from stools of chimpanzees experimentally infected with HAV was shown to possess multiple buoyant densities in CsCl gradients. The greatest proportion of HAV was most frequently found at a buoyant density of 1.32-1.34 g/cm3, however, large proportions of HAV were also frequently found at higher densities, including 1.36-1.37, 1.40-1.42, and 1.45-1.48 g/cm3. These findings are consistent with the notion that HAV may be a parvovirus.
Asunto(s)
Hepatovirus/análisis , Animales , Centrifugación por Gradiente de Densidad , Enterovirus/clasificación , Heces/microbiología , Hepatovirus/clasificación , Humanos , Pan troglodytes , Parvoviridae/clasificaciónRESUMEN
Experimental infection of two chimpanzees with the Phoenix Antigen strain of HAV resulted in the cyclic excretion of virus particles on days 9-11, 14-15, and 20-21 postinoculation. Isopycnic banding in CsCl of stool suspensions prepared from 9-11; 14-15; and 17, 19, 21 dav stool pools revealed multiple buoyant densities for the associated HAV particles. Hollow HAV particles found in the 9-11 day pool banded primarily at a buoyant density of 1.30 g/cm3. HAV in the 14-15 day stool banded bimodally in a CsCl gradient, with antigen peaks at buoyant densities of 1.29 and 1.33 g/cm3. HAV in the days 17, 19, 21 stool pool also banded bimodally in a CsCl gradient; however, the antigen peaks occurred at buoyant densities of 1.33 and 1.40 g/cm3.
Asunto(s)
Hepatitis A/microbiología , Hepatovirus , Animales , Antígenos Virales/análisis , Centrifugación por Gradiente de Densidad , Modelos Animales de Enfermedad , Heces/microbiología , Hepatovirus/inmunología , Hepatovirus/aislamiento & purificación , Pan troglodytes , PeriodicidadRESUMEN
Well-defined, core-like structures were visualized in hepatitis A virus particles by a modified microelectron microscopy technique.
Asunto(s)
Hepatovirus/ultraestructura , Cápside , Microscopía ElectrónicaRESUMEN
Sera of 103 carriers of hepatitis B surface antigen were assayed for e-antigen and anti-e. Twenty-four were e-antigen-positive, 31 anti-e-positive, and 48 had neither detectable (e-negative). Aminotransferases were elevated in 75% of the e-antigen-positive carriers compared with 25% of e-negative carriers (P less than 0.001) and 13% of anti-e-positive carriers (P less than 0.001). Serum DNA polymerase activity was significantly higher in the e-antigen-positive carriers than in carriers without e-antigen. Dane particles were shown in 10 of 12 carriers with e-antigen, compared with one of 12 e-negative carriers (P less than 0.0003) and none of 12 anti-e-positive carriers (P less than 0.00003). These results suggest that ongoing hepatitis B viral replication is more active in e-antigen-positive carriers than in carriers without e-antigen, a finding that may help explain the high prevalence of chronic active hepatitis described in these individuals.
Asunto(s)
Adolescente , Adulto , Alanina Transaminasa/sangre , Anticuerpos Antivirales/análisis , Aspartato Aminotransferasas/sangre , Niño , Preescolar , Femenino , Humanos , Masculino , Persona de Mediana Edad , Replicación ViralRESUMEN
During investigation of a food-borne outbreak of hepatitis A among university students in a southwestern metropolitan community, immune electron microscopic examination of a concentrated stool suspension pooled from seven acutely ill individuals revealed viruslike particles 17-nm in diameter. These particles were initially coated by antibody contained in the convalescent-phase serum of one of the ill students as well ad by antibody in convalescent plasma of a prison volunteer originally infected with the MS-1 strain of hepatitis A virus. Rises in titer of antibody to this particle were demonstrated by immune electron microscopy in acute and convalescent sera from student patients as well as in pre-inoculation and convalescent sera from the prison volunteer. Two chimpanzees inoculated intravenously with the concentrated preparation of pooled human stools developed viral hepatitis. During acute illness their feces contained particles morphologically identical to those in the inoculum. These findings represent the first reported recovery of the presumed etiologic agent of hepatitis A from a naturally occurring community outbreak of disease in the United States.
Asunto(s)
Brotes de Enfermedades , Hepatitis A/etiología , Enfermedad Aguda , Animales , Anticuerpos Antivirales/análisis , Reacciones Antígeno-Anticuerpo , Antígenos Virales/análisis , Arizona , Heces/microbiología , Contaminación de Alimentos , Hepatitis A/inmunología , Humanos , Cuerpos de Inclusión Viral , Microscopía Electrónica , Pan troglodytesRESUMEN
Virus-like particles shown to be associated with hepatitis A infection were recently visualized by immune electron microscopy in human and chimpanzee acute-phase fecal extracts. Agarose gel filtration of concentrated chimpanzee fecal extracts containing those 27-nm diameter particles separated more than 99% of the high molecular weight fecal impurities into two major peaks as determined by absorbance at 280 and 260 nm. The hepatitis A-associated particles were found between these two peaks by both immune electron microscopy and a new immunoradiometric assay.
Asunto(s)
Heces/microbiología , Hepatitis A/veterinaria , Hepatovirus/aislamiento & purificación , Pan troglodytes/microbiología , Animales , Reacciones Antígeno-Anticuerpo , Cromatografía en Agarosa , Hepatitis A/microbiología , Microscopía ElectrónicaRESUMEN
Transformed chimpanzee lymphocytes were examined to determine whether they would support the replication of hepatitis B virus. After 5 months, no hepatitis B virus, hepatitis B surface antigen, or antibody to hepatitis B surface antigen was detected.
Asunto(s)
Virus de la Hepatitis B/inmunología , Linfocitos/inmunología , Replicación Viral , Animales , Portador Sano , Células Cultivadas , Pruebas de Fijación del Complemento , Técnica del Anticuerpo Fluorescente , Hepatitis B/inmunología , Anticuerpos contra la Hepatitis B/análisis , Antígenos de la Hepatitis B/análisis , Herpesvirus Humano 4/inmunología , Activación de Linfocitos , Linfocitos/ultraestructura , Microscopía Electrónica , Pan troglodytes , RadioinmunoensayoAsunto(s)
Infecciones por Echovirus/epidemiología , Meningitis Viral/epidemiología , Adolescente , Adulto , Alaska , Líquido Cefalorraquídeo/microbiología , Niño , Preescolar , Infecciones por Echovirus/líquido cefalorraquídeo , Infecciones por Echovirus/microbiología , Enterovirus Humano B/inmunología , Enterovirus Humano B/aislamiento & purificación , Heces/microbiología , Femenino , Vivienda , Humanos , Masculino , Meningitis Viral/líquido cefalorraquídeo , Meningitis Viral/microbiología , Pruebas de Neutralización , Faringe/microbiología , Saneamiento , Abastecimiento de AguaAsunto(s)
Enfermedades Fetales/microbiología , Mononucleosis Infecciosa/microbiología , Complicaciones Infecciosas del Embarazo/microbiología , Aborto Terapéutico , Líquido Amniótico/microbiología , Anticuerpos/análisis , Pruebas de Fijación del Complemento , Citomegalovirus/aislamiento & purificación , Femenino , Enfermedades Fetales/inmunología , Feto/microbiología , Edad Gestacional , Pruebas de Hemaglutinación , Humanos , Inmunodifusión , Inmunoglobulina A/análisis , Inmunoglobulina G/análisis , Inmunoglobulina M/análisis , Mononucleosis Infecciosa/inmunología , Riñón/microbiología , Hígado/microbiología , Intercambio Materno-Fetal , Embarazo , Complicaciones Infecciosas del Embarazo/inmunología , Orina/microbiologíaRESUMEN
Thirty-four neonatal beagles were inoculated with cells obtained originally from 2 adult beagles, 1 with lymphocytic leukemia, the other with generalized malignant lymphoma. Most of the puppies were irradiated before inoculation; a few were inoculated in utero and were not irradiated. Of the 11 puppies inoculated with buffy coat cells of the blood from the leukemic donor, 2 developed leukemia. Of the 23 pups inoculated with cells of lymph nodes from the spontaneous lymphoma or positive passages, 13 developed lymphoma. Time of measurable onset of leukemia or lymphoma in the recipients varied from 12-60 days after inoculation. One passage was achieved with the leukemia and three serial passages with the lymphoma. A female karyotype was obtained from a neoplastic lymph node in a male recipient of the lymphoma. Since the donor was female, this suggests that the passages of the lymphoma were transplants.