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Virulence of pathogenic and non-pathogenic zymodemes of Entamoeba histolytica (Indian strains) in guinea-pigs.

Guinea-pigs were inoculated with Entamoeba histolytica strains isolated from cases of amoebic liver abscess, amoebic dysentery and asymptomatic cyst passers and the strains were classified in zymodemes by isoenzyme electrophoresis. It was observed that certain non-pathogenic zymodemes were potentially pathogenic for the guinea-pigs. The relationship of zymodemes and virulence in laboratory animals is discussed.

Amebiasis

Protection against establishment of latent infections in mice immunized with a non-pathogenic herpes simplex virus mutant and reinfected with the pathogenic parental strain.

Immunization of hairless mice with a TK-, ACVr, non-pathogenic herpes simplex virus (HSV) type 1 mutant protected the mice against reinfection with lethal doses of the parental pathogenic HSV strain. The protection conferred by the mutant against the establishment of latency after reinfection with the parental strain was dependent on the site of reinfection; after reinfection at the same site, only 2% of mice became latently infected, compared to 32% after reinfection at a distant site. When inoculation with the mutant was done at two different sites, single reinfections at any site led to the establishment of latency in 4% of the mice. The mutant by itself was almost completely latency-negative: only 4.5% of the mice developed latency in trigeminal ganglia and 2.3% in the spinal ganglia. The rate of mutant-induced latent infections is partly related to the dose of the virus; however, lower doses of the mutant may not colonize the ganglia, and therefore fail to protect against challenge infections.

Animals

Sequence analysis and relationships between meningococcal class 3 serotype proteins and other porins from pathogenic and non-pathogenic Neisseria species.

The presence of highly conserved regions within previously determined porin gene sequences from Neisseria meningitidis and Neisseria gonorrhoeae permitted the construction of oligonucleotide primers for PCR amplification of other neisserial porin genes. Although two separate porin genes (porA and porB) are present in N. meningitidis only a single fragment, corresponding to porB, could be amplified from this species. The amplified porB genes from four different meningococcal serotypes, which express the class 3 outer membrane protein, were sequenced. Amplified fragments corresponding to porin genes from N. lactamica and N. sicca were also sequenced. In common with the known neisserial porins, models of the organisation of the predicted proteins indicated trans-membrane structures with eight surface exposed loops. In the meningococcal class 3 proteins the main regions of sequence variation, which must be responsible for serotype specificity, were located on loops 5 and 7. A phylogenetic analysis of the family of porins from the Neisseria confirmed the close relationship of the meningococcal class 3 protein with the gonococcal PIA protein, while the gonococcal PIB protein was shown to be closely related to the N. lactamica porin. The close relationship seen between porins of the pathogenic and non-pathogenic Neisseriae identified no obvious virulence-associated regions in the proteins, but did suggest that the current nomenclature for neisserial porin genes may need reviewing.

Alleles

Ribosomal DNA sequences in the differentiation of pathogenic and non-pathogenic isolates of Entamoeba histolytica.

Recombinant ribosomal DNA sequences were amplified by PCR and used as probes to perform a fingerprint analysis of total DNA from different Entamoeba histolytica isolates. RFLPs obtained with one of the probes, R-1, support previous proposals that pathogenic and non-pathogenic E. histolytica are closely related, yet genotypically distinct. Another probe, R-2, while not distinguishing between the two forms of E. hystolytica, was able to differentiate between them and E. moshkovskii, which has morphologically identical cysts and trophozoites. A third probe, BR-1, identified strain-specific RFLPs.

Animals

Asymptomatic intestinal colonization by pathogenic Entamoeba histolytica in amebic liver abscess: prevalence, response to therapy, and pathogenic potential.

Since the application of isoenzyme electrophoresis to the study of Entamoeba histolytica, the prevalence and natural history of asymptomatic intestinal colonization in patients with amebic liver abscess (ALA) has not been addressed. We prospectively evaluated this enteric phase in 50 patients with ALA, using two dosage regimens of metronidazole. The overall prevalence of asymptomatic colonization was 72% (36/50). All these isolates, without exception, proved to express pathogenic zymodemes. Despite a 100% clinical response of the hepatic lesions, failure to eradicate the organism from the bowel occurred in 20 of these 36 subjects. During longitudinal posttreatment surveillance, three carriers returned with second bouts of invasive disease: one with dysentery and two with liver abscesses. Thus, in patients with ALA, there is a high prevalence of intestinal colonization with exclusively pathogenic strains, and treatment with metronidazole frequently results in a continued carrier state. These carriers have a propensity for developing recurrent invasive disease and constitute a public health hazard.

Adult

Fate of pathogenic and non-pathogenic Escherichia coli strains in two fermented milk products.

The growth and survival of pathogenic and non-pathogenic strains of Escherichia coli was determined in traditionally fermented pasteurized and unpasteurized milk and in Lacto, an industrially fermented milk. Each milk treatment was incubated at 20 degrees C for 24 h and then stored at either 20 degrees C or 5 degrees C for 96 h. Lacto inhibited all the three E. coli strains. Two strains could not be recovered and the third survived only in very low numbers after 24 h storage of Lacto at both 20 degrees C and 5 degrees C. All three E. coli strains survived and multiplied to maximum cell numbers in the range 10(7)-10(9)/ml during traditional fermentation of unpasteurized milk. Cell numbers decreased to 10(3)-10(6) and 10(2)-10(5) during storage of the fermented product at 20 degrees C and 5 degrees C respectively. Higher maximum numbers, 10(9)-10(10), of the three strains of E. coli were attained during traditional fermentation of pasteurized milk. The numbers decreased to 10(5)-10(8) and 10(4)-10(7) during storage of the fermented product at 20 degrees C and 5 degrees C respectively. Generally, fewer E. coli survived when the fermented milk products were stored at refrigeration temperature.

Animals

Pathogenic mycoplasmas: cultivation and vertebrate pathogenicity of a new spiroplasma.

A spiroplasma recovered from allantoic fluids of chick embryos infected with the tick-derived suckling mouse cataract agent was grown in continuous passage on a new artificial culture medium. The cultured organisms induced typical ocular and other disease symptoms in susceptible animals, and were reisolated from involved host tissues. Although spiroplasmas have been previously recognized as plant and insect pathogens, this is the first spiroplasma shown to multiply at 37 degrees C and to be pathogenic for vertebrates.

Animals

Activity of four cephalosporin antibiotics in vitro against bovine udder pathogens and pathogenic bacteria isolated from newborn calves.

The in vitro activity of chephaloridine, cephalexin, cefatrizine (BL-S640), and cephapirin (BL-P-1322) was evaluated by the serial dilution method against pathogenic gram-positive and gram-negative bacteria isolated from bovine udders and neonatal calf diseases. Cephapirin showed the comparatively greatest activity against the most common streptococcal species associated with bovine mastitis, whereas cephaloridine exhibited the best activity against Staphylococcus aureus. Cefatrizine was more active than the other cephalosporins against the gram-negative bacteria studied. In general, the minimal bactericidal concentration of each cephalosporin was two- to fourfold lower than the comparative value reported in the literature against the same type of pathogen of human origin.

Animals

The 96-kilodalton antigen as an integral membrane protein in pathogenic Entamoeba histolytica: potential differences in pathogenic and nonpathogenic isolates.

A surface antigen (EH-96) of Entamoeba histolytica was demonstrated to be a plasma membrane antigen by immunoprecipitation of metabolically 35S-labeled antigen from live trophozoites, Triton X-114 detergent extracts, and plasma membrane-enriched fractions prepared by concanavalin A membrane stabilization and differential centrifugation. In addition, the antigen was localized to the plasma membrane by electron microscopy with colloidal gold. Antigen from E. histolytica strains immunoprecipitated with specific immunoglobulin M (IgM) or IgG2b monoclonal antibody was identical by one-dimensional peptide mapping with N-chlorosuccinimide. Additionally, antigen from different axenically cultivated amebae was demonstrated to be identical by N-chlorosuccinimide peptide mapping, as were peptide maps of IgG and IgM monoclonal antibody-purified antigen. The 96-kilodalton (kDa) surface antigen was identified on four axenically cultivated pathogenic isolates and on three polyxenically cultivated pathogenic isolates (zymodeme II) of E. histolytica but was absent or present in lesser quantity on six nonpathogenic polyxenically cultivated isolates. The 96-kDa antigen was detected in liver abscess fluid from four patients with amebic abscesses by enzyme-linked immunosorbent assay (ELISA) and immunoprecipitation. Two-dimensional gel electrophoresis profiles of the 96-kDa antigen purified from abscess material or from polyxenically cultivated trophozoites demonstrated that the antigens were related to the 96-kDa antigen found in axenically cultivated organisms.

Animals

An experimentally pathogenic Bacillus species. II. The pathogenicity of the organism for mice.

The pathogenic effects produced in mice by intraperitoneal (i.p.) injection of a Bacillus species (OSU 372) are presented. This organism belongs to group 3 of the genus, and members of this group have not heretofore been shown to be pathogenic for mice even under experimental conditions. However, this organism is capable of producing a fatal involvement in doses which are not considered to be overwhelming. The mean lethal dose (LD50) of the organism for 20-25 g mice by the i.p. route is about 1 times 10-8 bacteria/mouse. A rapid drop in body temperature along with severe dehydration were noted in infected animals, and hematologic studies indicated that leukopenia and hemoconcentration also occurred. Although a transient septicemia developed, the bacteria could not be recovered from the tissues of fatally infected mice after a certain point in time. Results prevented indicate that the animals died of hypovolemic shock. A possible parallel with human bacillary infection is drawn.

Animals

[Studies of the incidence of healthy carriers of "pathogenic strains" or potentially pathogenic strains in the hospital environment].

An investigation has been carried out on healthy carriers of "positive Staphylococcus coagulase" and "occasional pathogenic" strains among kitchen, canteen and dispensary staff in the IVth Geriatric Division and First Aid Clinic of the Bologna "M. Malpighi" Hospital. All stains of "positive Staphylococcus coagulase" and "occasional pathogens" considered (Pseudomonas aeruginosa and Citrobacter) were put through antibiotic and chemotherapeutic sensitivity tests. The results are reported in detail.

Anti-Bacterial Agents

Pathogenic and non-pathogenic Naegleria and Acanthamoeba spp.: a new autochthonous isolate from an Italian thermal area.

We performed an epidemiological survey of 17 thermal baths and the same number of mud-basins. This study aimed to ascertain the presence and incidence of small free-living amoebae, particularly species and/or strains of Naegleria and Acanthamoeba spp., occasional etiological agents of fatal meningoencephalitis and/or ocular infections in man. Over 51 samples of water and mud incubated at 37 degrees C and at 45 degrees C, 34 (66.7%) became positive at 37 degrees C and 33 (64.7%) at 45 degrees C. We isolated 7 (6%) strains of Naegleria spp., 6 (5.2%) of Acanthamoeba spp., 39 (33.6%) of Vahlkampfia spp., 28 (24.1%) of Hartmannella spp. and 36 (31.1%) strains of other species of free-living amoebae. 4 strains of Naegleria spp. and 6 of Acanthamoeba spp. proved pathogenic both in vivo, after experimental infection (meningoencephalitis) in the albino mouse, and in vitro, having previously contaminated monolayers of Vero cell line (cytopathic effect). Within the isolated pathogenic strains of Naegleria spp., a new strain of N. australiensis s.sp. italica was typified from an immunochemical point of view. This should be added to previous isolations reported by us.

Amoeba

[Aggravation of experimental dysentery in mice and acceleration of the process of multiplication of pathogenic and conditionally pathogenic bacteria under the effect of yeast RNA].

Simultaneous infection of mice with Sh. flexneri subtype 2a, culture, with various doses of RNA sodium salt (NaRNA) or administration of the preparation 3 hours prior to or 6, 12, and 18 hours after the infection was accompanied by aggravation of the experimental infection in mice. This was expressed in a greater number of animals which contracted the disease and died, and in increased blood and peritoneal exudate microbiol density. The same processes were noted in mice infected with equal doses of shigella cultures passaged four times through broth with NaRNA, in comparison with the animals infected with shigellae passaged through broth alone. Incubation of a number of pathogenic and conditionally pathogenic microbes in broth with NaRNA was accompanied by a significant increase in the bacterial biomass volume; at the same time other conditionally patoogenic microbes or even different strains of the same type of the microbes reacted to NaRNA but weakly. A possibility of realization of all of these processes in vivo and their influence on the origination of bacterial complications and development of the infection is supposed.

Animals

Isoenzyme patterns of pathogenic and non-pathogenic Naegleria spp. using agarose isoelectric focusing.

Using agarose isoelectric focusing, the isoenzyme patterns of 7 different enzymes were compared in 52 Naegleria strains. The pathogenic N. fowleri was found the most homogeneous species. N. lovaniensis seems to be constituted of different types which form nevertheless a cohesive group. Within N. gruberi, large interstrain band variations were found in almost all enzyme systems. A re-examination of the taxonomic position of this species may therefore be taken into consideration. High temperature strains from Australia were confirmed to be different from N. lovaniensis. Members of a new pathogenic Naegleria sp., N. australiensis, seem to occur in Europe. Large thermophilic strains with many large pores in the cysts show identical zymograms and may constitute a new species or genus.

Acid Phosphatase

The pathogenicity of Salmonella mbandaka in specific pathogen free rabbits.

Salmonella mbandaka was isolated from the lungs of three rabbits that developed a peracute disease and died three to eight weeks following extensive experimental manipulation. To examine the pathogenicity of this rare serotype, clinically normal, S. mbandaka-carrier rabbits were subjected to either corticosteroid immunosuppression alone or corticosteroid immunosuppression with a minor surgical procedure. Acute or peracute salmonellosis developed in one of two rabbits immunosuppressed only, and in three of four rabbits immunosuppressed and surgically manipulated. Stressful experimental procedures may precipitate episodes of clinical salmonellosis in rabbits harboring this organism. Salmonella mbandaka, should be considered a zoonotic agent in the laboratory environment.

Animals

Pathogenic activities of live cells and extracellular products of the fish pathogen Pasteurella piscicida.

The pathobiological activities in vivo and in vitro of live cells and extracellular products (ECP) of eleven Pasteurella piscicida strains of different origin were examined. Infectivity trials showed that P. piscicida did not possess strict host specificity since the majority of the isolates were virulent for gilthead seabream, rainbow trout and turbot, with LD50 values ranging between 10(3) and 10(6) live cells. However, none of the strains tested were pathogenic for mice (LD50 > 10(8) cells)). In addition, the ECP were strongly toxic for fish (LD50 ranging from 1.0 to 4.6 micrograms protein per g fish), which clearly demonstrates their important role in the pathogenesis of pasteurellosis. All the ECP samples were cytotoxic for fish and homoiothermic cell lines, possessed notable phospholipase activity and displayed haemolytic activity for sheep, salmon and turbot erythrocytes (but not for trout erythrocytes). However, the production of proteolytic enzymes differed among the P. piscicida strains. Although no strain displayed elastase activity, five isolates (the Japanese and Italian strains) hydrolysed casein and gelatin. All these biological activities in vivo and in vitro were lost after heat treatment (100 degrees C for 10 min). The general enzymic patterns of both live cells and ECP evaluated by the API-ZYM system also revealed some variation among the P. piscicida isolates. Generally, whole cells showed a wider range of enzymic activities than ECP. The results presented here are important for the selection of strains in the development of effective polyvalent pasteurellosis vaccines containing both whole cells and ECP.

Animals

[Murine model for experimental studies on attenuated mutants of a Pasteurella multocida strain with pathogenicity for the calf. 1. Isolation and biological characterization of streptomycin-dependent mutants (Sm-d) and their examination for immunogenicity in comparison with thermally inactivated pathogens].

An experimental study was conducted into the immunisation of mice to a strain of Pasteurella multocida with pathogenicity for calf. Here are the results: (a) Stable Sm-d mutants were isolated. (b) The genetic stability of this mutant type was established by unsubstantiable reversion up to the magnitude of 10(-8) or - in cases in which 20 Sm-id revertants maximum occurred in 10(-8) Sm-d germs--by prolongation of the generation period of these suppressor mutants which can be defined also as minus mutants. (c) Stable attenuation of selected Sm-d mutants has been established in mice by intraperitoneal application of 108 germs. (d) Live germs with restricted reproduction exhibited more immunogenicity to thermally inactivated antigen.

Animals