Serodiagnosis of deep-seated fungal infections.
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Biomedical subjects
Publications and source records attributed to H P Seeliger.
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Listeria monocytogenes was isolated from the brain of a goat, which was euthanized due to listeriosis. A few weeks later a similar subtype of L. monocytogenes was isolated from an on-farm manufactured fresh cheese which did not contain any milk from the goat which had suffered from listeriosis. A similar subtype was also found on 1 of the shelves in the refrigerator where cheeses were stored. Prior to the onset of listeriosis, 1 fresh cheese had been made of milk from the actual goat, which may have excreted L. monocytogenes in her milk. Thus, the cheese made of this milk may have contaminated the shelves in the refrigerator which then has served as a Listeria reservoir for new cheeses during several weeks.
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A 40-year-old man with grade II sarcoidosis went on a two-week visit to Ecuador. Afterwards he was treated with low-dose corticosteroids because of slight deterioration on the X-ray film. Unilateral hilar enlargement followed by bouts of high fever and a greatly increased erythrocyte sedimentation rate, suggested an acute exacerbation of the sarcoidosis and was, therefore, treated with an increase in steroid dosage plus azathioprine. Bronchoalveolar lavage fluid and tissue samples from lung, bone-marrow and spleen were suggestive of an infection with Histoplasma capsulatum. There was now bilateral hilar enlargement with right upper lobe infiltration, marked hepatosplenomegaly and thrombocytopenia (17,000/microliters), but serological tests remained negative. The fulminant course with dissemination could not be arrested despite administration of fluconazole (400 mg/d) and amphotericin B (total dosage 1.14 g). Histoplasma capsulatum was cultured from lung and spleen tissue post-mortem.
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Pathogenicity testing of 10-day-old fertilized hen's eggs infected with Listeria species demonstrated that L. monocytogenes isolated from cheese and human samples caused death in 100% of the injected chicken embryos within 96 h. In contrast, L. innocua isolated from cheese was fatal for only 17% of the infected embryos. All embryos infected with L. seeligeri survived. The results confirmed statements of previous authors about the pathogenicity of L. monocytogenes, regardless of its origin.
Aldo Castellani was one of those pioneers who initiated breath-taking discoveries and progress in medical sciences during the early years of this century with great personal engagement and courage. His striking findings, such as the so-called Absorption-Test, the introduction of multiple vaccination, or the exploration of sleeping sickness and the discovery of Trichophyton rubrum, were achieved before he was even 35 years old. He received acknowledgement as a physician and lecturer. Many of his disciples and coworkers became first rate scientists, owing a lot to his encouraging personality. Castellani's straight forward character endeared him to many famous contemporaries. His uncompromising nationalism for Italy on the other hand arouse astonishment and aversion. His involvement in three wars again and again, rendered scientific researches and work difficult.
Listeriosis usually manifests at the extremes of age, during pregnancy or among immunocompromised individuals as an acute meningoencephalitis with or without associated septicemia. Localized infections are rare. We investigated the incidence of Listeria monocytogenes cholecystitis using the data of the Listeria reference laboratory of the University of Würzburg/FRG. Out of 467 culture proven L.m-infections in the years 1986 and 1987, two cases were localized infections of the gall bladder. Although studies of the microbiology of gallbladder infections have not demonstrated the recovery of Listeria monocytogenes as a pathogen in acute or chronic cases of cholecystitis, these two cases substantiate the existence of this rare form of listeriosis as an entity and support the theory of the importance of the gastrointestinal tract in the pathogenesis of Listeria infections.
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Although apparently observed before, the history of listeriosis dates back approximately 60 years. First known as a cause of epidemics and sporadic cases in some 50 species of animals, the disease appears now with increased frequency among human populations at risk. The causative agent Listeria monocytogenes is primarily a psychrophilic soil-borne bacterium with a wide pathogenic potential thus provoking primarily septicemia, meningitis and intrauterine infections. Recent observations indicate certain types of food being the principle vehicle for transmission of human listeriosis. This would parallel the epizootic situation in domestic animals. Further studies of the mechanisms leading to clinical and subclinical infections are just as necessary as reliable methods to determine the immunity status of individuals at risk.
Listeriosis in humans is a rare disease, which, however, is known to be epidemic and endemic. The prognosis has remained unsatisfactory up to today, the fatality being at least 10% and often considerably higher depending on the clinical features of the disease and the patient's age. Three population groups are at risk: pregnant women, fetuses and newborn infants. Furthermore, immunosuppression in older patients due to disease, therapy, or age also plays a role. The incidence of Listeria infections in patients over 45 is clearly increasing. Due to the nature of the pathogen (in vivo bactericidal concentrations of antibiotics are often not attainable; intracellular growth) a high dosage of ampicillin is recommended. Although the present therapeutic possibilities are not satisfactory, a combination of ampicillin and an aminoglycoside appears to be the best therapy at present. Other combinations such as rifampicin and beta-lactam antibiotics have exhibited in vitro antagonism. The preferred therapy, ampicillin, can only be recommended with reservations because it is not optimally effective.
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296 strains of Listeria monocytogenes have been submitted for confirmation and further studies to the Listeria Reference Laboratory at the Institute of Hygiene in Würzburg, Federal Republic of Germany. They have been isolated between 1969 and 1985 from human cases in the Federal Republic and West Berlin. The results of an analysis of the respective cases are presented here on the basis of questionnaires. A steady increase of Listeria isolations has been noted during the past few years. The largest number of strains originated from the district Nordrhein-Westfalen which has the largest population of all German federal districts. The highest incidence calculated from the number of strains and total population was found for the district of Saarland. There was no predominance detectable of listeriosis among the rural population. 60% of the strains came from patients living in urban areas. In 6 out of 21 cases of neonatal listeriosis at least one of the parents was engaged in professional medical services. The most frequently observed clinical entities were meningitis (41.14%) and septicaemia (36.00%). Among the newborns septicaemia was predominant with 40.90%. Listeriosis during pregnancy and among newborns was connected with 53.57% of all strains isolated. The questionnaire revealed for the underlying conditions in 35% and in 24% of cases malignancies and organ transplantations, respectively. There was no increased rate of predominance found among the elderly. However, 50% of all strains isolated originated from newborns. The sex distribution was almost equal; 52.56% from males of the total number of isolates and 51.22% from males among the newborns. 67% of these which had a meningitis after the first weeks of life were male. There was no seasonal incidence detectable. Serovar 4b was found in 66.22% of all isolates tested. 295 strains produced acid from rhamnose and alpha-methyl-d-mannose, but not from d-xylose and were thus typical for L. monocytogenes. One strain needs further studies.
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Listeriolysin was isolated from target rabbit erythrocyte membranes after lysis of the cells with partially purified toxin derived from a culture supernatant of Listeria ivanovii. The membrane form of the toxin exhibited properties similar to those previously found for streptolysin O. Detergent-solubilized, delipidated listeriolysin was found to comprise a heterogeneous population of partially and fully circularized, amphiphilic oligomers whose embedment within the lipid bilayer generated large transmembrane pores. The molecular weight of the toxin monomer was estimated to be 55,000 to 60,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Immunological cross-reactions between the toxin and streptolysin O were demonstrable by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. An immunoblot assay for detecting listeriolysin in agar-incorporated, lysed erythrocyte membranes was developed, and 28 defined, clinical isolates of Listeria monocytogenes were examined for toxin production. These isolates caused beta-hemolysis on the agar plates and had previously been regarded as listeriolysin producers. However, we found that only two isolates produced genuine listeriolysin, since the sensitive immunoblot assay entirely failed to detect the toxin in all other cases. We excluded that this finding derived from proteolytic degradation of membrane-bound toxin. Thus, the great majority of human pathogenic Listeria strains appear to produce one or several hemolysins that are immunologically and, by inference, molecularly distinct from the streptolysin O-related listeriolysin. We propose that the streptolysin O-related toxin be designated alpha-listeriolysin and that the other hemolysin(s) be termed beta-listeriolysin.