[Influence of lyophilization on composition and antigenic power of Eberthella typhosa and Salmonella paratyphi B].
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Freeze Drying↗
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[Experimental ingestion of salmonella (eberthella) typhosa].
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Salmonella↗
[Therapeutic effect of sulfobenzyl-penicillin (SB-PC) on the concentration of SB-PC in human bile (author's transl)].
Sulfobenzyl-penicillin (SB-PC) is currently being investigated for use in man. The purpose of this study was to evaluate effect of SB-PC on Salmonella typhosa and biliary excretion of SB-PC in disease of the biliary tract. 1) Clinically, typhoid carriers without cholelithiasis were initiated with 4.0 g/day of SB-PC. Stool and bile became negative for Salmonella typhosa 14 days after initial treatment. 2) In typhoid carriers with cholelithiasis, Salmonella typhosa were not isolated from bile, wall of the gallbladder and surface of gallstone, but were isolated from nuclei of gallstones. The treatment of typhoid carriers with cholelithiasis may belong to a most difficult problem. 3) Biliary excretion of SB-PC in the patient given a single dose of 2.0 g/day im. was markedly dependent on characteristics of the patient, situation of external drainage and volume of bile excretion. High concentration in bile in some patient was 298 mug/ml at 3 hours after administration. 4) In intravenous administration of single dose of 6.0 g. maximum concentration in bile was about 2,000 mug/ml at 2 hours after administration and bactericidal concentration was obtained for resistant bacteria (Pseudomonas, Proteus, etc.) in biliary infection. 5) As side effects, pain and redness were infrequent after im. administration. Toxicity was not experienced in the patients injected intravenously.
Aged↗
Retention of bacterial lipopolysaccharide at the site of subcutaneous injection.
The tissue distribution of Klebsiella pneumoniae O3 lipopolysaccharide (KO3 LPS) was studied in mice injected subcutaneously (s.c.) or intraperitoneally (i.p.) with 125I-labeled KO3 LPS. Marked retention of KO3 LPS radioactivity could be found at the site of s.c. injection for several weeks. On the other hand, about 85% of the radioactivity rapidly disappeared from the peritoneal cavity within 6 h after i.p. injection. The long-term presence of KO3 LPS at the injection site was also supported by experiments with 51Cr-labeled KO3 LPS and immunoblotting and immunofluorescence staining methods. The R-form LPS lacking the O-specific polysaccharide chain of KO3 LPS and the lipid A fraction of KO3 LPS seemed to remain at the site in larger amounts and for longer times than KO3 LPS. There were no marked differences in the retention pattern at the injection site among KO3 LPS, Escherichia coli LPS, Salmonella typhosa LPS, and Salmonella enteritidis LPS. However, much less radioactivity accumulated in the livers and spleens of mice injected with either KO3 LPS or S. typhosa LPS compared with the other LPS preparations. It was suggested that retention of LPS at the site of s.c. injection may play an important role in the development of various biological actions of s.c. injected LPS.
Animals↗
Chromosomal location of ribosomal protein cistrons determined by intergeneric bacterial mating.
Intergeneric mating between Escherichia coli and Salmonella typhosa was used to locate at least three 30S ribosomal proteins near the streptomycin locus in the region of 54 to 66 min of the E. coli map. This procedure utilizes differences in the electrophoretic patterns of 30S ribosomal protein of the parents. The results show that cistrons for 30S proteins of E. coli can replace those of S. typhosa in the Salmonella genome. Moreover, in a diploid hybrid with a Salmonella endogenote and an E. coli exogenote, both sets of cistrons are expressed.
Bacterial Proteins↗
Fever, antipyretics and serum iron levels in rabbits.
The intracerebroventricular (i.c.v.) injection of bacterial endotoxin (Shigella dysenteriae) produces a rise in deep-body temperature in rabbits. The increase in body temperature was partially inhibited by polymyxin B (P), and completely suppressed by ketoprofen (K). The systemic injection of bacterial endotoxins (Shigella dysenteriae or Salmonella typhosa) elicited an elevation of body temperature in rabbits which was partially blocked by polymyxin B and completely antagonized by ketoprofen. Serum iron concentration fell in rabbits treated with Salmonella typhosa endotoxin and rose after injections of ketoprofen, polymyxin B, indomethacin and ketoprofen plus endotoxin (En). However, the total increases in serum iron levels were higher after antipyretics than after ketoprofen plus endotoxin treatment. These results show that the antipyretics antagonized the rise in body temperature and the fall in serum iron concentration induced by bacterial endotoxin and all the drugs used in this study to induce antipyresis increased serum iron levels.
Animals↗
Intracerebroventricular taurine in rabbits: effects of normal body temperature, endotoxin fever and hyperthermia produced by PGE1 and amphetamine.
1. Intracerebroventricular (I.C.V.) injections of taurine into rabbits resting at an ambient temperature (Ta) of 10 degrees or 23 degrees C caused hypothermia but at 30 degrees C ambient temperature, rectal temperature was unchanged. 2. An I.C.V. bolus of 0=5 mg taurine immediately followed by a slow infusion of taurine (0-01--0-2 mg/min) into rabbits at 23 degrees C ambient temperature caused sedation and peripheral vasodilation and blocked the febrile response to Salmonella typhosa endotoxin (1 microng/kg i.v.). Sustained fevers, characteristic of fevers caused by central administration of pyrogens, developed after taurine infusions were stopped. Control infusions of taurine at the same rates in the same rabbits when they were afebrile had little effect on rectal temperature. 3. An I.C.V. injection of 0-5 mg taurine reduced the hyperthermia caused by prostaglandin E1 (PGE1; 2 microng) given I.C.V. A dose of 5-0 mg not only blocked PGE1 hyperthermia but also caused marked hypothermia. 4. Bilateral injections of taurine into the preoptic/anterior hypothalamic region, at sites where injections of Salmonella typhosa endotoxin caused long-lasting fevers, had no effect on rectal temperature. Similar injections into the reticular substance of the medulla oblongata, in the region believed to be concerned with a secondary temperature control function, were also without effect on body temperature. 5. Taurine (0-5 and 5-0 mg, I.C.V.) had no consistent effect on hyperthermia induced by amphetamine (2 mg/kg, I.V.) 6. We conclude that the hypothermic effect of taurine is not due to an action on the central neurone pool or pools concerned with the integrative control of thermoregulatory effectors. This amino acid appears to inhibit neuronal activity in efferent pathways which control peripheral vasomotor tone and heat production and to depress the level of arousal. Taurine delays the onset and extends the duration of endotoxin-induced fever, perhaps by two separate action: by inhibiting activity in central thermoregulatory pathways and by promoting accumulation of endogenous pyrogen in the brain.
Amphetamines↗
Sequences of synthesis of gamma-1 macroglobulin and gamma-2 globulin antibodies during primary and secondary responses to proteins, salmonella antigens, and phage.
The nature of the antibodies produced by the rabbit during the primary and secondary responses to T(2) phage, proteins, and the O and H antigens of Salmonella typhosa has been determined. Immune sera have been fractionated by zone electrophoresis, sucrose density ultracentrifugation, and anion exchange chromatography. The resulting fractions have been assayed by phage neutralization or hemagglutination (antisera to proteins) or bacterial agglutination. In confirmation and extension of earlier work from this laboratory, the primary response to these antigens, with the exception of the O antigen of the Salmonella, included the early synthesis of 19S, gamma-1 globulin antibody, and the later synthesis of 7S, gamma-2 globulin antibody. The primary response to the O antigen consisted of the synthesis of only a macroglobulin agglutinin. The secondary response to the proteins, including the H antigen of the Salmonella, comprised the early synthesis of large amounts of the 7S gamma-2 globulin antibody to the same level attained during the primary response. The secondary response to the phage consisted in the synthesis of 7S, gamma-2 globulin antibody alone. Treatment of the macroglobulin phage-neutralizing antibody with mercaptoethanol resulted in complete loss of its neutralizing activity. A working hypothesis to explain these observations was presented. A salient feature of this hypothesis was the suggestion that different cells synthesized the two distinct molecular forms of antibody. The significance of the sequential synthesis of the two forms of antibody is not known. It was proposed that the system for synthesis of macroglobulin antibody is an auxiliary system for antibody synthesis, perhaps the first to develop phylogenetically and ontogenetically. It is felt that the present observations indicate a clear-cut qualitative distinction between the primary and secondary responses to immunization whereby these responses might be identified in various experimental situations. It is also felt that these findings with the primary and secondary responses to various antigens in the rabbit may be of widespread occurrence in nature among a variety of species.
Animals↗
INFLUENCE OF NEOMYCIN AND INGESTED ENDOTOXIN IN THE PATHOGENESIS OF CHOLINE DEFICIENCY CIRRHOSIS IN THE ADULT RAT.
Two groups of adult rats fed a choline-deficient diet supplemented with neomycin in their drinking water for 250 or 350 days were protected against the development of liver fibrosis and cirrhosis. At the termination of the study these animals weighed more than others not receiving neomycin. This difference in weight did not appear to be caused by a growth-promoting effect of neomycin but rather reflected the increased severity of liver disease and a resultant weight loss in animals not receiving neomycin. Protection by neomycin was cancelled when Salmonella typhosa endotoxin was added to the drinking water. It was concluded that the protective effect of neomycin was mediated by an alteration in the intestinal microflora resulting in a reduction in the numbers of organisms contributing to intraluminal endotoxin. In the presence of choline deficiency, absorption of intraluminal endotoxin may contribute to the development of fibrosis and cirrhosis.
Animals↗
CHROMOSOME TRANSFER KINETICS OF SALMONELLA HFR STRAINS.
Johnson, E. M. (Walter Reed Army Institute of Research, Washington, D.C.), Stanley Falkow, and L. S. Baron. Chromosome transfer kinetics of Salmonella Hfr strains. J. Bacteriol. 88:395-400. 1964.-The kinetics of chromosome transfer of an Hfr strain of Salmonella typhosa and an Hfr strain of S. typhimurium were examined in interrupted matings with multiply auxotrophic S. typhimurium recipients. The S. typhosa Hfr, TD-7, was found to transfer the pro-A, met-A, arg (A, C, F, or H), and ile markers at 8, 32, 36, and 51 min, respectively, after contact with the recipient strain. Comparison of these entry times with those of the analogous Escherichia coli Hfr P4X-6 for the same markers showed the gene order to be identical. However, the TD-7 entry times were considerably extended over those of P4X-6, which transfers these markers of E. coli F(-) strains at, respectively, 5, 20, 22.5, and 28 min. A similar extension of the entry times was noted with the S. typhimurium Hfr, SR-305, which transfers the markers in the reverse order, ile-met-A-pro-A, at 3 to 4, 18, and 46 min, respectively. Examination of P4X-6/Salmonella Hfr entry time ratios showed them to be constant at 0.63 for the earlier markers transferred by both TD-7 and SR-305. These data suggest that the physical length of the Salmonella chromosome is the same as that of E. coli, and that the rate of chromosome transfer of the Salmonella Hfr strains to S. typhimurium recipients is only 0.63 that of P4X-6 to E. coli F(-) strains under the same physical conditions.
Amino Acids↗
DETECTION AND PERSISTENCE OF VI ANTIGEN IN TISSUES OF ACTIVELY IMMUNIZED MICE.
Gaines, Sidney (Walter Reed Army Institute of Research, Washington, D.C.), Julius A. Currie, and Joseph G. Tully. Detection and persistence of Vi antigen in tissues of actively immunized mice. J. Bacteriol. 89:776-781. 1965.-The presence, distribution, and persistence of Vi antigen in mouse tissue was determined by means of active immunization tests with tissue extracts. Mice were injected intraperitoneally with purified Vi antigen or Vi-containing bacilli. At appropriate intervals, animals were killed, and saline extracts of their tissues were prepared. Mice were immunized with these extracts and challenged 6 days later with 10 ld(50) of Salmonella typhosa Ty2. Protection was afforded by tissue extracts from Vi-injected mice, but not by normal tissue extracts. That the immunizing capacity of tissue extracts from Vi-injected mice was attributable to Vi antigen was affirmed by the demonstration that these extracts stimulated the production of Vi antibody in mice, coated erythrocytes for agglutination by Vi antiserum, and inhibited agglutination of Vi-sensitized red blood cells by known Vi antisera. Vi antigen could be detected in the liver and spleen of mice injected with as little as 1 mug. In mice given 150 mug, the antigen was still present in liver tissue 231 days later.
Animals↗
Relation of structure to function in bacterial O antigens. I. Isolation methods.
Nowotny, Anna M. (City of Hope Medical Center, Duarte, Calif.), Scott Thomas, Olga S. Duron, and Alois Nowotny. Relation of structure to function in bacterial O antigens. I. Isolation methods. J. Bacteriol. 85:418-426. 1963.-Six isolation methods, four previously described and two reported herein, were compared for the extraction of endotoxic O antigens from three different enterobacteria: Serratia marcescens Bizio, Salmonella typhosa O901, and Escherichia coli K-12 lambda. Large variations were observed in the toxin yield, in the serologically reactive material yield, and in the nucleic acid-releasing abilities of the six methods when applied to the different bacterial strains. Chemical analyses of the toxic O antigens included quantitative determination of the amino acids, long- and short-chain carboxylic acids, and carbohydrates. Comparisons of the nucleic acid content were also made. Although the chemical composition of the preparations was quite different, no correlation could be found between the percentage of the above components and the lethal toxicity or the serological reactivity. It was concluded that the structural elements or properties responsible for these biological activities cannot be detected by gross chemical analyses.
Antigens↗
MECHANISMS OF ENDOTOXIN TOLERANCE. II. RELATIONSHIP BETWEEN ENDOTOXIN TOLERANCE AND RETICULOENDOTHELIAL SYSTEM PHAGOCYTIC ACTIVITY IN MAN.
Healthy male volunteers were rendered tolerant to the pyrogenic and toxic activities of bacterial endotoxin by daily intravenous injections. Five subjects were given 0.5 microg Salmonella typhosa endotoxin for 7 days; four subjects were given Pseudomonas endotoxin, increasing over a period of 30 days from 25 to 250 microg. Reticuloendothelial system (RES) phagocytic activity was assessed by serial measurements of the clearance of I(131)-labeled aggregated human serum albumin. In no subject was an increase in RES phagocytic activity detectable. Such negative findings could not be attributed to decreased RES blood flow.-Additional studies on the pyrogenic responses of man to various schedules of endotoxin administration revealed: (a) Hyperreactivity of some subjects to a second injection of endotoxin administered 24 hours after the initial dose; (b) prevention of such hyperreactivity by plasma from donors tolerant to a heterologous endotoxin, but not from normal donors; (c) reduced reactivity to a second injection of endotoxin given 7 days after the initial dose; (d) reversal of induced tolerance by administration of half the dose of endotoxin followed 2 hours later by the second half; (e) reversal of induced tolerance 24 hours after administration of a heterologous endotoxin; (f) enhanced dermal reactivity to endotoxin induced inflammation during tolerance. The observations are consistent with the hypothesis that tolerance to the pyrogenic activity of endotoxin in man is not based upon generalized enhancement of RES phagocytic activity or exhaustion of host reactivity but rather involves the participation of specific antibody which assists the RES in the clearance and inactivation of the endotoxin molecule.
Antigen-Antibody Reactions↗
Action of bacterial endotoxin and lipid A on mitochondrial enzyme activities of cells in culture and subcellular fractions.
Escherichia coli O127:B8 lipopolysaccharide (LPS), prepared by the Westphal procedure, caused a marked decrease in the activities of mitochondrial malate dehydrogenase, succinate dehydrogenase, and adenylate kinase in African green monkey kidney (Vero) cells and primary cultures of mouse liver cells within 2 h after exposure to 10 micrograms of LPS/ml of culture medium. These three enzyme activities leaked into the supernatant fraction, and cytochrome oxidase activity was lost from the mouse liver mitochondrial particulate fraction within 45 min after exposure to 10 micrograms of LPS/mg of protein. Loss malate dehydrogenase activity from isolated mitochondria was also accelerated by LPS from E. coli O26:B6 (Boivin preparation) or Salmonella typhosa O901 (Westphal preparation), and by lipid A from Salmonella minnesota or Shigella sonnei. In addition, LPS and lipid A inhibited state 3 respiration by isolated mitochondria with attendant loss of respiratory control, but adenosine 5'-diphosphate/O ratios were relatively unchanged. Impaired mitochondrial function is an early event after exposure to biologically relevant amounts of LPS or lipid A.
Adenylate Kinase↗
Responses of baboons to traditionally pyrogenic agents.
It is not clear whether baboons develop fever in response to endotoxin or other pyrogens. We injected various pyrogens intravenously in 12 unrestrained baboons (Papio ursinus) and measured their body temperature using intra-abdominal radiotelemeters. Serum iron concentration was also measured. The baboons developed fever after injection of killed Staphylococcus aureus (5 X 10(7) organisms/kg). No significant fever was measured after injection of lipopolysaccharide (Salmonella typhosa) (0.1, 8, 40, and 100 micrograms/kg), bovine serum albumin (4 mg/kg), killed Salmonella minnesota (5 X 10(7) organisms/kg), and killed Salmonella typhi (5 X 10(7) organisms/kg). A significant decrease in serum iron concentration was found only after injection of S. aureus and lipopolysaccharide, 100 micrograms/kg. The phagocytic synthesis of interleukin-1 following pyrogen stimulation in baboons and some other primates appears to differ from that in man and in nonprimates.
Animals↗
Role of typhoid antigens in protection and pathogenicity for mice.
Swiss white mice immunized with acetone-killed vaccines prepared from strains of Salmonella typhosa, S. typhimurium, and mouse-virulent S. typhimurium hybrids which had acquired, by conjugal genetic transfer, the S. typhosa antigens 9, Vi, and d were challenged with the S. typhimurium hybrids and with the S. typhimurium parent strain. The results of these experiments suggested that the Salmonella somatic antigens were important in conferring protection against death in this system. The S. typhosa Vi antigen did not appear to play any significant role in this protection. The S. typhimurium hybrids employed in these studies did not show any loss of mouse virulence as the consequence of acquisition of various combinations of the S. typhosa somatic, flagella, or Vi antigens, nor did S. typhosa hybrids which had acquired the somatic antigen of S. typhimurium show any increase in mouse virulence.
Animals↗
Behavior of coliphage lambda in hybrids between Escherichia coli and Salmonella.
Salmonella typhosa hybrids able to adsorb lambda were obtained by mating S. typhosa recipients with Escherichia coli K-12 donors. After adsorption of wild-type lambda to these S. typhosa hybrids, no plaques or infective centers could be detected. E. coli K-12 gal(+) genes carried by the defective phage lambdadg were transduced to S. typhosa hybrids with HFT lysates derived from E. coli heterogenotes. The lysogenic state which resulted in the S. typhosa hybrids after gal(+) transduction differed from that of E. coli. Ability to produce lambda, initially present, was permanently segregated by transductants of the S. typhosa hybrid. S. typhosa lysogens did not lyse upon treatment for phage induction with mitomycin C, ultraviolet light, or heat in the case of thermoinducible lambda. A further difference in the behavior of lambda in Salmonella hybrids was the absence of zygotic induction of the prophage when transferred from E. coli K-12 donors to S. typhosa. A new lambda mutant class, capable of forming plaques on S. typhosa hybrids refractory to wild-type lambda, was isolated at low frequency by plating lambda on S. typhosa hybrid WR4254. Such mutants have been designated as lambdasx, and a mutant allele of lambdasx was located between the P and Q genes of the lambda chromosome. Plaques were formed also on the S. typhosa hybrid host with a series of lambda(i21) hybrid phages which contain the N gene of phage 21. The significance of these results in terms of Salmonella species as hosts for lambda is discussed.
Adsorption↗
Enhanced toxicity for mice of 6-mercaptopurine with bacterial endotoxin.
The toxicity of 6-mercaptopurine was potentiated by 2 mg of either Escherichia coli 026:B6 B endotoxin or Salmonella typhosa 0901 W endotoxin per kg. Nonlethal doses of heat-killed, gram-negative bacteria were also capable of potentiating the lethality of 6-mercaptopurine (6-MP). Salmonella minnesota S, the wild-type strain, and S. minnesota Re 595, a mutant containing only the lipid A and 2-keto-3-deoxyoctonate moiety of the endotoxin molecule, exhibited the same capability to enhance the toxic action of 6-MP. Endotoxin (lipopolysaccharide [LPS]) did not affect the clearance of 6-MP from the circulation, but did alter its apparent metabolism as indicated by blood levels of a metabolite, 6-thiouric acid. The concentration of blood urea nitrogen (BUN) in mice 18 h after injection of 100 mg of 6-MP per kg simultaneously with 2 mg of LPS per kg was significantly elevated over normal values. However, these BUN values were significantly less than those resulting from the administration of one mean lethal dose of either agent. The clearance from the circulation of the gram-negative organism E. coli HB, or the gram-positive organism Staphylococcus epidermidis S, was not affected by 6-MP. Endotoxin had no effect on the clearance of S. epidermidis S, but inhibited that of E. coli HB. When 6-MP and LPS were administered simultaneously with either bacterial species, only the clearance of E. coli HB was inhibited. Mice were protected from the lethality associated with combinations of 6-MP and LPS by (i) prior treatment with phenobarbital, (ii) caffeine, (iii) methylprednisolone, and (iv) polymyxin B sulfate. With the exception of caffeine, each regimen protected mice against the lethal effects of 400 mg of 6-MP per kg, and methylprednisolone or polymyxin B protected mice against 8 mg of LPS per kg.
Animals↗