Studies on pathogenic leptospirae. VI. Effect of tween 80 on the growth of Leptospira canicola and Leptospira icterohaemorrhagiae.
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Stalheim, O. H. V. (University of Wisconsin, Madison), and J. B. Wilson. Cultivation of leptospirae. I. Nutrition of Leptospira canicola. J. Bacteriol. 88:48-54. 1964.-The nutrition of Leptospira canicola was investigated by use of synthetic media of suitable ionic strength. At an incubation temperature of 30 C, the minimal components were calcium, iron, magnesium, and ammonium ions, thiamine, and a fatty acid source; barium and strontium replaced calcium. Aspartic acid, glutamic acid, or methionine stimulated the rate and amount of growth; the best growth occurred in medium containing additional amino acids. Additions of cyanocobalamin or biotin permitted growth at 37 C. The stimulatory effects of added cyanocobalamin, biotin, pyridoxine, pantothenate, lipoic acid, or nicotinic acid were additive at 37 C, but not at 30 C. Fatty acids containing 14, 16, 17, or 18 carbon atoms supported growth; linoleic and linolenic acids were toxic. Glyceryl monooleate or trioleate, or Tween 40, 60, or 80 supported moderate to good growth; a mixture of monoolein and Tween 60, or Tweens 60 and 80 supported the best growth. Ten strains of L. canicola cultivated in a synthetic medium containing Tweens 60 and 80 attained cellular densities per ml of 10(7) to 4.0 x 10(7) organisms. L. canicola cells, resuspended in medium containing oleic-1-C(14) acid, incorporated label primarily into cellular lipids; a lesser amount was located in the protein fraction, and only trace amounts were found in the nucleic acid fraction. The rate of incorporation was not affected by added sodium acetate. L. canicola was found to have fatty acid decarboxylase activity.
Pregnant hamsters were inoculated with Leptospira canicola by an intraperitoneal route. Grossly, the livers were enlarged and pale yellow. On light microscopic examination, disruption of the hepatic cords, as well as granulation of hepatocyte cytoplasm, was evident. On electron microscopic examination, the livers had evidence of severe cellular degradation as early as post-inoculation hour (PIH) 24. At PIH 96, pathologic changes were more pronounced. The rough endoplasmic reticulum was swollen and there was membrane disruption. These changes were not observed in the livers of control animals. In addition, livers of the fetuses at PIH 72 exhibited severe degenerative changes.
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In an attempt to define the pathogenetic mechanisms involved in acute interstitial nephritis due to Leptospira canicola the kidneys of 14 dogs were subjected to histological immunofluorescence and elution studies. Leptospiral antigen was found in three sites within the kidney; as discrete organisms in the proximal tubules, as granular deposits in the cytoplasm of macrophages and in large extracellular clumps in the interstitium. Deposits of IgG were also detected in the latter site and in plasma cells found in the surrounding cellular infiltrate. Anti-leptospiral antibody, mainly of the IgG class, was detected in renal eluates using both agglutination-lysis and indirect immunofluorescence. From these findings it was considered that the cellular infiltrates found in the kidneys of dogs suffering from acute leptospiral nephritis have two main functions; firstly, the local production of anti-leptospiral antibody and, secondly, phagocytosis.
An electron microscopic study on the histopathological changes in the kidneys of pregnant hamsters resulting from infection with Leptospira canicola was undertaken. The experiment revealed damage to the glomeruli as well as the tubules. The severity of these lesions increased with the progress of the disease and examination of kidney sections showed leptospires in the tissue. It is suggested that the stress of pregnancy has some influence on the severity of the lesions seen in this disease.
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This report describes the epizootiology, clinical presentation, diagnosis and treatment of an outbreak of leptospirosis caused by Leptospira canicola in feedlot calves. The infection appeared to be of high morbidity with a cumulative clinical incidence of 15.6%, cumulative subclinical incidence of 39% and high mortality (8.3%). Clinical disease was diagnosed in 4-8-month-old calves, while subclinical infection occurred in 9-12-month-old calves. Subclinical infection was based on serological evidence only. The zoonotic aspects of the infection are emphasised.
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A man was admitted to the Johannesburg Hospital with a history of fever, diarrhoea, and dry cough for four days. He began to produce bloodstained sputum and was found to have severe arterial hypoxaemia. Radiography showed widespread opacification over both lung fields, and the clinical and haemodynamic features were consistent with the adult respiratory distress syndrome. Serology for Leptospira canicola was positive. Despite antibiotics, supportive therapy, and ventilation the patient died. Necropsy excluded cardiac disease. This case shows that leptospirosis may cause the adult respiratory distress syndrome.
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