A Preliminary Study of Ammonia Production by Corynebacterium renale and some other Pathogenic Bacteria

Abstract

SUMMARY:Urease is a constituent enzyme of Corynebactmiurn rende and appears to account for the bulk of its arnmonia production. C. renale also contains an arginase and some amino-acid deaminases, but the former has not been fully characterized. Bovine urine supports the growth of a small inocdum of C. renale for a limited time but after growth has reached a maximum it diminishes rapidly and the ammonia and pH values increase. C. renale also contains uricase but its precise significance has not yet been determined.Of the other bacteria studied, C. owis has urease activity similar to C. renak, and C. p y 0 . g~ a stronger initial arginase. C. equi contains no appreciable urease or arghwe, although it tends to form ammonia from glutamine.Corynebacterium renale is the cause of a specific cystitis and pyelonephritis in cattle; the lesions in natural infections in cattle and in experimental infections in mice and rabbits after intravenous inoculation are localized in the medulla of the kidney (Lovell, 1946; Lovell & Cotchin, 1946; Feenstra, Thorp & Gray, 1949). This selective localization may, in part, be explained by the metabolic and enzyme activities of the organism, and a study of its ammonia-producing capacity was made because of the old observation that C. r e d produces ammonia when grown in urine. A preliminary test was made to confirm this by growing a strain of C. r e d in bovine urine and in peptone water containing 1 % (w/v).urea. After 48 hr. incubation at 37" flasks of uninoculated media and those in which the organism had been grown were filtered through Seitx filters and the filtrates examined for free ammonia, urea and creatine. The results are given in Table 1, and show that in bovine urine and in a urea medium C. renale breaks down the urea with the formation of free ammonia.The creatine content of the urine remained constant, but largely diminished in the urea medium probably because of the peptone present.The ammonia-producing capacity of C. re& was therefore examined and a few comparative tests made with C. ovis, C. equi, C. pyogenes and Bacteriz.bm coli. METHODSThe methods employed were : (a) Washed suspensions of C. renak prepared from growth on agar slopes were seeded into sterile urine, peptone water containing 1 yo urea, hydrolysed peptone water and peptone water. The urine was sterilized by Seitz-filtration, and the hydrolysed peptone was prepared from a stock solution which consisted of a mixture of 100 g. of Bactopeptone and 300 ml. of cone. hydrochloric acid (A.R.) which had been boiled under a reflux condenser for 48 hr. after which most of the acid had been removed by vacuum distillation; the solution was neutralized to pH 7 by NaOH, diluted to give approximately 1 yo NaCl;

Urea Utilization by Leptospira

“…Secondly, there is some similarity between the overall renal pathology attributed to leptospiral infections and that associated with other types of bacterially induced renal disease. Since some of the bacteria capable of proliferation in the kidney medulla and causing renal damage (5,6,15,(22)(23)(24) possess urease, the demonstration of urease activity in representatives of at least two pathogenic leptospiral serotypes suggests a possible relationship between this enzymatic activity and the disease process itself. Studies on the role of urease in pyelonephritis caused by P. mirabilis have suggested that the alkalinity of the kidney tissues resulting from the presence of ammonia produced via the hydrolysis of urea may lead to local necrosis and the precipitation of salts in the renal tubule (6).…”

“…Nonspecific factors such as inhibition of phagocytic response by high salt and urea concentrations (9, 27,29) and probably inactivation of complement by ammonia (3) contribute to the general susceptibility of kidney tissue to infection. However, studies with Corynebacterium renale (22,23), Proteus mirabilis (4, 6,24), and various other bacteria (5,8,15) have suggested that the possession of a highly active urease, which is associated with the rapid hydrolysis of urea and ability to utilize high concentrations of urea, appears to influence the persistence of each of these organisms in the kidney medulla of the respective hosts and the severity of the kidney damage inflicted. Previ-I Present address: Laboratory Unit, Georgia Department of Human Resources, Atlanta, Ga.…”

Coryneform Bacteria in Poultry Deep Litter

“…This is unlike the litter isolates which are nonhaemolytic, but it is not certain how constant is the feature. Carne (1939) reported haemolysis in about two-thirds of his ovine strains and strains which were apparently nonhaemolytic could produce haemolytic colonies on subsequent culture; Morse (1949) states that his urease positive strains are nonhaemolytic, and Lovell & Harvey (1950) give no information on the haemolytic ability of their strains which hydrolyse urea. Coynebacterium renale, another strain reported as being urease positive (Lovell, 1946;Morse, 1948Morse, , 1949Lovell & Harvey, 1950), produces in litmus milk an alkaline reaction with clearing, whereas the litter isolates produce either no change or reduction with or without rennet-like curdling.…”

“…Carne (1939) reported haemolysis in about two-thirds of his ovine strains and strains which were apparently nonhaemolytic could produce haemolytic colonies on subsequent culture; Morse (1949) states that his urease positive strains are nonhaemolytic, and Lovell & Harvey (1950) give no information on the haemolytic ability of their strains which hydrolyse urea. Coynebacterium renale, another strain reported as being urease positive (Lovell, 1946;Morse, 1948Morse, , 1949Lovell & Harvey, 1950), produces in litmus milk an alkaline reaction with clearing, whereas the litter isolates produce either no change or reduction with or without rennet-like curdling. Morphological and colonial differences were noted when comparing cultures of C. ovis and C. renale with the litter strains; the animal pathogens are further distinctive in showing no action on a variety of organic acids whereas representative strains of the litter isolates are quite active in this respect on the medium used.…”