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Application of oxygen-enriched aeration in the production of bacitracin by Bacillus licheniformis.

The physiological effects of controlling the dissolved oxygen tension at 0.01, 0.02, and 0.05 atm by the use of oxygen-enriched aeration were investigated during growth and bacitracin production by Bacillus licheniformis ATCC 10716. Up to a 2.35-fold increase in the final antibiotic yield and a 4-fold increase in the rate of bacitracin synthesis were observed in response to O(2)-enriched aeration. The increase in antibiotic production was accompanied by increased respiratory activity and an increase in the specific productivity of the culture from 1.3 to 3.6 g of antibiotic per g of cell mass produced. Oxygen enrichment of the aeration decreased medium carbohydrate uptake and the maximum specific growth rate of B. licheniformis from 0.6 h(-1) to as low as 0.15 h(-1), depending upon the level of enrichment and the conditions of oxygen transfer rate (impeller speed). The response of this culture to O(2) enrichment suggests that this method of controlling the dissolved oxygen tension for antibiotic-producing cultures may simulate conditions that would occur if the carbon source were fed slowly, as is often employed to optimize antibiotic production. Analysis of the biologically active bacitracins produced by B. licheniformis ATCC 10716 suggested that the ratio of biologically active peptides was not changed by O(2) enrichment, nor were any new biologically active compounds formed.

Bacillus↗

Effect of rifampin and bacitracin on nasal carriers of Staphylococcus aureus.

Anterior nares cultures from 664 hospital personnel identified 165 (24.8%) as carriers of Staphylococcus aureus. Persistent carriers (17.8%) were identified and randomly assigned to one of four treatment groups: rifampin (600 mg once a day for 5 days), bacitracin ointment (topically applied three times a day for 10 days), combined rifampin and bacitracin, and control (no treatment). Bacitracin ointment was ineffective in eliminating S. aureus from the anterior nares and had a postreatment carrier rate equal to the control rate. Rifampin therapy caused a highly significant reduction (79%) in carriage; however, combined therapy was not as effective as treatment with rifampin alone. Of 132 strains of staphylococci isolated before and after treatment, all were susceptible to less than 0.016 micrograms of rifampin per ml. This study demonstrates that rifampin may be an effective antistaphylococcal antibiotic and could be used to control the carrier state in high-risk situations.

Adult↗

Distribution of streptococcal groups in clinical specimens with evaluation of bacitracin screening.

During a 2-year period, 4,968 strains of beta-hemolytic streptococci were examined for the clinical source distribution and bacitracin sensitivity of each group. In the upper respiratory tract, groups A (51.7%) and C (20.4%) accounted for most of the isolates, and in wounds and exudates group A (79.1%) made up most of the isolates. Group B (71.2%) was the major component of isolates from the genitorinary tract and, while composing 29.3% of the lower respiratory tract isolates, competed with group A (18.8%) and the nongroupables (22.8%) for supremacy. Bacitracin screening showed that 0.5% of group A streptococci were resistant, and sensitive non-group A isolates were group B (2.6%), group C (6.0%), group G (8.0%), and the nongroupables (2.2%). It was found that those groups which were most predominant in wounds and the upper respiratory tract gave the highest rate of false positives with bacitracin, whereas the predominant group of the genitourinary tract gave the lowest rate of false positives.

Bacitracin↗

Purification of the extracellular protease of Bacillus licheniformis and its inhibition by bacitracin.

Sporulating cells of Bacillus licheniformis excrete three seryl proteases that are of similar size, 28,000 daltons, but of different charge at pH 6. The peptide antibiotic bactracin is released from the cells at the same time and exists, in part, as a bacitracin-protease complex that is stable throughout chromatographic procedures employed in enzyme purification. However, preextraction of crude protease with CHCl3 and subsequent gel filtration effect separation of the antibiotic and the enzyme. Three purified, bacitracin-free proteases, designated CMC I, CMC II, and CMC III and whose ratios of total activity are 1:3.7:10.3, respectively, are obtained by chromatography on carboxymethyl cellulose. The major component, CMC III, is inhibited by commercial bacitracin at near-physiological concentrations of the antibiotic.

Bacillus↗

Mechanism of bacitracin resistance in gram-negative bacteria that synthesize exopolysaccharides.

Four representative species from three genera of gram-negative bacteria that secrete exopolysaccharides acquired resistance to the antibiotic bacitracin by stopping synthesis of the exopolysaccharide. Xanthomonas campestris, Sphingomonas strains S-88 and NW11, and Escherichia coli K-12 secrete xanthan gum, sphingans S-88 and NW11, and colanic acid, respectively. The gumD gene in X. campestris is required to attach glucose-P to C55-isoprenyl phosphate, the first step in the assembly of xanthan. A recombinant plasmid carrying the gumD gene of X. campestris restored polysaccharide synthesis to bacitracin-resistant exopolysaccharide-negative mutants of X. campestris and Sphingomonas strains. Similarly, a newly cloned gene (spsB) from strain S-88 restored xanthan synthesis to the same X. campestris mutants. However, the intergeneric complementation did not extend to mutants of E. coli that were both resistant to bacitracin and nonproducers of colanic acid. The genetic results also suggest mechanisms for assembling the sphingans which have commercial potential as gelling and viscosifying agents.

Bacitracin↗

Lack of reliability of primary grouping of beta-hemolytic streptococci by culture of throat swabs with streptocult supplemented with bacitracin disks in general practice.

Fifty-eight general practitioners took throat swabs from 434 patients with sore throats. Office cultures were performed on Streptocult supplemented with bacitracin disks in an attempt to carry out primary grouping of beta-hemolytic streptococci (BHS). In 424 cases the findings were compared with those obtained in a microbiological laboratory. Streptocult showed a sensitivity of 75% and a specificity of 84% in the detection of BHS. The office-performed grouping procedure of the correctly detected BHS, however, only had a sensitivity of 65% and a specificity of 87%. Overall, as many as 45% of the patients with BHS group A were misdiagnosed. The unsatisfactory results obtained with primary grouping of BHS may be due partly to incorrect determinations of the diameter of the inhibition zone around the bacitracin disks and partly to an inappropriate choice of breakpoint. It is concluded that cultures of throat swabs on Streptocult in general practice should not be accompanied by attempts to carry out primary grouping with bacitracin disks. A laboratory investigation showed that incubation at room temperature for 48 h and at 35 degrees C for 24 h gave identical BHS positivity rates.

Bacitracin↗

Variations in bacitracin susceptibility observed in Staphylococcus and Micrococcus species.

Bacitracin susceptibility was evaluated as a laboratory method to differentiate staphylococci from micrococci. A total of 317 staphylococcal isolates and 108 micrococcal isolates were each tested for susceptibility to bacitracin by a disk-diffusion method using disks of three different potencies (0.04, 2.0, and 10.0 U) and a broth dilution method to obtain MICs. When a growth inhibition zone diameter breakpoint of greater than 10 mm was used to establish susceptibility with a 0.04-U disk, all micrococci were bacitracin susceptible and 94.6% of the staphylococci were resistant. Testing with disks of higher potency did not improve the specificity of the disk-diffusion method.

Bacitracin↗

Bacitracin-resistant clone of Streptococcus pyogenes isolated from pharyngitis patients in Belgium.

We report 16 bacitracin-resistant Streptococcus pyogenes isolates recovered from pharyngitis patients in Belgium, 14 of which belonged to a particular emm type (emm28). All 16 isolates were constitutively resistant to macrolides and carried erm(B). The emergence of a bacitracin-resistant S. pyogenes clone raises questions about the continued reliability of bacitracin susceptibility testing for S. pyogenes identification.

Anti-Bacterial Agents↗

Bacitracin and coagglutination for grouping of beta-hemolytic streptococci.

Bacitracin may be used for presumptive differentiation of group A from other beta-hemolytic streptococci. Whatever criteria are used for the test, a small number of erroneous results will be obtained. With the 1,161 strains studied in this laboratory, using a zone of 10 mm or greater as indicative of group A was better than using Maxted's original criterion of any zone of inhibition for group A. Co-agglutination is a preferable alternative to bacitracin testing, providing a grouping result as quickly as the bacitracin test but with the advantage of giving a definite grouping result. With the 247 strains studied so far, coagglutination results have correlated exactly with results of conventional grouping by a precipitin method.

Bacitracin↗

A nuclear magnetic resonance study of the metal binding sites in bacitracin.

Carbon-13 nuclear magnetic resonance spectroscopy has been used to identify sites in bacitracin which bind Cu2+ and Mn2+. Results are presented which implicate the free carboxyl groups of the aspartic and glutamic acid residues and the imidazole ring of the histidine residue as metal complexation sites between pH 6 and 8. Evidence is presented which also indicates that the thiazoline ring of bacitracin binds Mn2+. Bacitracin does not bind Cu2+ or Mn2+ at pH values of 2.5 or less.

Bacitracin↗

Studies of angiotensin I converting enzyme: effects of kinins, bacitracin, gamma-aminobutyric and epsilon-aminocaproic acids, and related compounds on substrate binding and catalysis in vitro.

Bradykinin and 22 of its analogs were evaluated for their abilities to inhibit the hydrolysis of [3H]hippurylglycylglycine by purified porcine kidney angiotensin I converting enzyme. The mean inhibitory concentration (IC50) for bradykinin was 1.2 +/- 0.2 X 10(-6) M. Except for Ile-Ser-bradykinin and [Sar4]-bradykinin, none of the kinin analogs were more potent in this regard than bradykinin. Bacitracin, gamma-aminobutyric acid, epsilon-aminocaproic acid, and structurally related compounds were also tested. The IC50 value for bacitracin was 1.9 +/- 0.4 X 10(-4) M, gamma-aminobutyric acid, 83.4 +/- 7.2 mM, and for epsilon-aminocaproic acid, 7.0 +/- 1.4 mM. Compounds were also evaluated for their abilities to prevent 125I-labelled [Tyr1]-kallidin binding to angiotensin I converting enzyme inhibited by EDTA. The IC50 values for bradykinin, bacitracin, gamma-aminobutyric acid, and epsilon-aminocaproic acid were 1.6 +/- 0.3 X 10(-8) M, 2.6 +/- 0.9 X 10(-6) M, greater than 291 mM, and 13.2 +/- 3.9 mM, respectively.

Aminocaproates↗

Intraoperative anaphylaxis after irrigation with bacitracin: case report.

We report a case of anaphylaxis caused by irrigation with a bacitracin solution during lumbar laminectomy. The patient had been exposed to bacitracin during a previous anterior cervical discectomy. We recommend avoiding the use of irrigation solutions containing bacitracin in patients with previous systemic exposure to this antibiotic.

Aged↗

Does bacitracin have an absorption-enhancing effect in the intestine?

We studied the absorption enhancement effects of three types of protease inhibitors, aprotinin, bacitracin and soybean trypsin inhibitor, in the rat intestine. Of these protease inhibitors, bacitracin enhanced the absorption of FD-4 and phenol red from the rat small and large intestine without mucosal toxicity. Thus, it was suggested that bacitracin has not only a protease-inhibitory but also an absorption-enhancing capability.

Animals↗

Influence of bacitracin on microbial functions in the gastrointestinal tract of horses.

This study investigated the influence of zinc bacitracin on the intestinal flora of horses. The functionally active intestinal flora was examined in 6 horses during treatment with zinc bacitracin. Utilising gas chromatography, spectrophotometry, gel electrophoresis and paper chromatography, samples were analysed on biochemical markers reflecting the action of parts of the intestinal flora. The following 5 flora-related functions were studied in faecal samples and intestinal samples from different sections of the hindgut: conversion of cholesterol to coprostanol and of bilirubin to urobilinogens, degradation of mucin and of beta-aspartylglycine and inactivation of tryptic activity. Conversion to coprostanol, conversion to urobilinogens and degradation of mucin were affected by treatment of zinc bacitracin and conversion to coprostanol was most sensitive. All functions were normalised in a short time, in contrast to man and rats. Differences in environmental exposures are probably the reason for a more rapid normalisation of the intestinal flora functions in horses.

Animals↗

Comparison of dio-bacit, bacitracin-trimethoprim/ sulphamethoxazole and latex agglutination in the diagnosis of Group A beta-hemolytic streptococci.

Not only is Group A beta-hemolytic Streptococcus (GAS) the most frequent cause of bacterial pharyngitis, it is also the culprit in various skin and systemic infections, acute rheumatic fever, post streptococcal glomerulonephritis, and other disorders and complications. A new, ready-to-use media, Dio-Bacit, in a two section plate containing 5% sheep blood agar on one side and sheep blood agar with bacitracin (2 microg/ml) on the other was compared for its efficiency in identifying GAS with bacitracin and bacitracin + sulphamethaxazole / trimethoprim disk tests applied after isolation of beta-hemolytic colonies. We also used the latex-agglutination test as the gold standard method for differentiating GAS from streptococci belonging to other groups. Compared with the latex-agglutination test, we found the sensitivity and specificity of the Dio-Bacit method to be 92.0% and 96.9%, respectively. Dio-Bacit plates provide an easy and very useful way to identify GAS within one day, saving time, labor, and money for routine diagnostic microbiology laboratories.

Adult↗

Structural characterization of bacitracin components by Frit-fast atom bombardment (FAB) liquid chromatography/mass spectrometry (LC/MS).

The structural characterization of minor components of bacitracin (BC) complex was carried out using a technique of liquid chromatography/mass spectrometry (LC/MS). Satisfactory total ion current chromatogram of BC complex and excellent mass spectra of many components were given by Frit-fast atom bombardment (FAB) LC/MS analytical system, and the structures of 13 minor components could be proposed. The 13 minor components were classified into two groups, bacitracin A (BC-A) related components and bacitracin F (BC-F) related components depending on their common N-terminal moieties. The structures of BC-A related components and BC-F related components were the same as those of BC-A and BC-F, respectively, except that one to three of isoleucine and leucine residues are replaced by valines. The BC-F related components were degradation products of BC-A related components through the same degradation process as that of BC-A.

Amino Acid Sequence↗

Biosynthesis of bacitracin in solid-state fermentation by Bacillus licheniformis using defatted oil seed cakes as substrate.

Bacitracin is being imported in Pakistan involving substantial amount of foreign exchange for its incorporation in poultry feed. The cheap raw material for its production is readily available and cheap such as soybean meal, sunflower meal, wheat bran etc. Thus development of this technology in our country would result in saving a reasonable amount of foreign exchange by exploiting indigenous resources. The present study is concerned with the biosynthesis of antibiotic bacitracin in solid-state fermentation by Bacillus licheniformis on laboratory scale using defatted oil seed cakes of agricultural by-products as starting material for maximum production of the antibiotic Bacitracin. In solid-state fermentation, wheat bran, soybean meal, sunflower meal, rice hulls and their different combinations were used. The antibiotic activity, 48 hours after inoculation was 4375 i.u / gm when only soybean was used. However, maximum titre 4820 i.u / gm of antibiotic was obtained using wheat bran and soybean meal in ratio of 1:3.

Anti-Bacterial Agents↗

Effect of the insulin degradation inhibitors bacitracin and chloroquine on insulin binding data to H35 rat hepatoma cells.

The influence of correction for internalization and of inhibition degradation by the use of bacitracin or chloroquine on the insulin binding data of H35 rat hepatoma cells has been assessed. Internalized insulin represented about 50% of total cell bound insulin at tracer concentration and is not affected by bacitracin or chloroquine. The use of the insulin degradation inhibitors bacitracin and chloroquine resulted in Scatchard plots that were strikingly less curvilinear than control cells (as indicated by a decreased K1/K2 ratio (p less than 0.001) and Ke/Kf ratio (p less than 0.01)). In the two site model there was a significant increase in the number of high affinity binding sites concomitant with a significant decrease in the high affinity association constant; there was also a significant decrease in the number of the low affinity binding sites. The total number of binding sites did not change. For the negative cooperativity model the use of each drug resulted in a significant increase in the affinity constant of the filled site. These results indicate that insulin degradation induces negative cooperativity and that the results are inconsistent with the two site model.

Animals↗