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Paper chromatography is very helpful in the identification and comparison of antibiotics, but very few results from the application of this technique have been published so far. The awakening of interest in the search for antibiotics which are active against fungi pathogenic to plants and animals makes the investigator aware of the scant information that is available to help characterize such agents. A knowledge of the Rf values of the known antifungal agents would help advance the search for such therapeutic materials. Often these values, in conjunction with the biological inhibitory activities, are the only criteria by which one can determine whether the unknown agent appears to be a new material. When the Rf data are obtained for shake cultures, one can eliminate many previously described antibiotic-producing organisms from further study, thus allowing more research to be applied to antibiotics which are new and different. The studies reported in this paper should serve as a guide to the identification of some of the known antifungal antibiotics.

Publisher Summary This chapter discusses the principles of paper chromatography of antibiotics. It includes all steps of analysis beginning with the preparation of samples and papers, development techniques, detection methods, and means of documentation. Paper chromatography of the best-known groups of antibiotics is also presented in the chapter. General theoretical aspects of paper chromatography are not dealt with except for those that directly emerged from studies of antibiotics. The chapter concludes with a discussion of paper chromatographic classification and systematic analysis of antibiotics.

(1960). The Paper Disc Method for Determination of Bacterial Sensitivity to Antibiotics Studies on the Accuracy of the Technique. Scandinavian Journal of Clinical and Laboratory Investigation: Vol. 12, No. 4, pp. 408-413.

Antibiotic discs were prepared, using several several batches of papers meeting Food and Drug Administration specifications. The analysis of 1,152 zones of inhibition produced showed no performance differences among these batches. Other discs were prepared using papers of different grades. These produced large differences in performance. It is obvious, therefore, that the use of a specified disc paper is necessary for standardizing the performances of the products of various manufacturers and that reproducible results can be attained with the grade of paper specified.

An attempt to establish a systematic chromatographic analysis of antibiotics has been made. Sixty-two known antibiotics were distributed into five classes and into fourteen subclasses, according to their RF values in four principal solvent systems. Principles of the classification of antibiotics are given. For more detailed comparison and characterization of antibiotics further additional solvent systems were used. Additional solvent systems A, B, C, and D were used for class I, systems E, F, G, and H for classes II and III, systems I, J, K, and L were used for classes IV and V.

Antibiotic 136, a mixture of five similar basic antibiotics, has been partially resolved by chromatography on activated carbon. Component A appeared identical with streptothricin by paper chromatography, elemental analysis, infrared spectrum and animal toxicity. Fraction ABCD could not be distinguished by paper chromatography from pleocidin. Fraction CDE showed chemical properties similar to the mixture streptolin AB. Among the acid hydrolysis products of Antibiotic 136 chromatographic fractions, l -β-lysine, α- d -gulosamine, streptolidine, and N-guan-streptoldyl- d -gulosaminide have been identified. Component D difers from components A, B, and C by containing an imidazoline structural unit differing from streptolidine. The general properties of the mixture Antibiotic 136 indicate that its components are members of the streptothricin complex of antibiotics.

Six healthy men were given a fruit- and vegetable-free diet, and one man was given a pure sucrose-water diet and antibiotics for 4 days. Vanylglycol in urine was detected by paper chromatography and thin-layer chroma tography after enzymatic hydrolysis with glusulase (glucuronidase + arylsul fatase). Acid hydrolysis destroyed vanylglycol. Treatment with antibiotics and a sucrose diet did not change the vanylglycol excretion, while the mono amineoxidase inhibitor, alphamethyldopa, and reserpine decreased the excre tion. Disulfiram treatment increased the vanylglycol excretion.

Scopafungin (U-29,479) is a crystalline, nonpolyenic antimicrobial agent obtained from the culture broth of Streptomyces hygroscopicus var. enhygrus var. nova UC-2397. Scopafungin inhibits, in vitro, a variety of pathogenic fungi, yeasts, and gram-positive bacteria.

Lomofungin is a new antimicrobial agent obtained from the culture broth of Streptomyces lomondensis sp. n. UC-5022. Lomofungin is an acidic, olive-yellow, crystalline compound which inhibits, in vitro, a variety of pathogenic fungi, yeasts, and gram-positive and gram-negative bacteria.

Column chromatography on a strongly basic ion-exchange resin, with water as eluent, fractionated the gentamicin antibiotic complex; each fraction, representing different mixtures of antibiotics, further separated upon thin-layer chromatography. Sixteen antibiotics were clearly differentiated. Paper chromatography of the N-acetyl derivatives supplemted thin-layer chromatography, detecting many additional constituents of the complex. A number of important, basic, water-soluble antibiotics were chromatographically compared with the major gentamicin antibiotics.

A graphic method of predicting antibiotic yields in continuous flow reactors is presented and discussed using the novobiocin fermentation as a model process. Extension to other antibiotic fermentations and steroid bioconversions is emphasized. In the case of the novobiocin fermentation it was concluded that a combination of one growth stage and one or two antibiotic production stages would be the most economic reactor system to conduct such a fermentation on a continuous basis.

Four patients are reported in whom perforation of the colon followed exchange transfusion for haemolytic disease of the newborn. This association seems to be more than coincidental, and possibly the perforation is due to a vascular accident occurring as a mechanical result of the exchange transfusion. The insidious onset of colonic perforation may be recognized early by the passage of blood per rectum. There is no place for conservative treatment, and once the diagnosis has been made treatment must include broad-spectrum antibiotics and laparotomy.

A new antibiotic, gluconimycin, was isolated from Streptomyces AS 9. The systematic position of the organism is discussed. Gluconimycin has a polypeptide nature. It contains iron and gluconic acid in its molecule. Thus it has been classified as a member of sideromycins. Gluconimycin is considered from the fast moving type when chromatographed by butanolacetic acid-water. The antibiotic is active against Gram+ve, Gram-ve bacteria and some fungi. The antibiotic exerts high toxicity when injected in mice.

Antibiotic-resistant enterobacteria, the vast majority of which carried transmissible R factors, were isolated from 81 out of a total of 100 healthy infants. There was surprisingly little correlation between previous antibacterial therapy and the resistant enterobacterial flora.

The use of many different antibiotics to treat chest infection has led us to test the sensitivity of 68 strains of Haemophilus influenzae to 15 different compounds. These included established compounds such as ampicillin and tetracycline and newer agents such as cephalosporins and clindamycin. The minimum inhibitory concentrations of the compounds for H. influenzae were then compared with blood levels attained after the usual dose regimens. There has been a significant increase in tetracycline resistance in the last few years, but all strains were sensitive to ampicillin, chloramphenicol, sulphamethoxazole, and trimethoprim, Several antibiotics were found to be microbiologically unsuitable for treating H. influenzae infections.

Out of 200 infections due to Bacteroides fragilis occurring over a period of three years 133 were related to the intestinal tract, 55 to the genitourinary tract, and the remainder were in bedsores and ulcers; 56% occurred in patients undergoing major intestinal surgery.B. fragilis was isolated in pure culture from 56% of the infections. In mixed culture it was most commonly associated with Klebsiella and Enterobacter species. Other anaerobic bacteria were isolated in 9% of the mixed cultures.Altogether 131 (65.5%) of the patients recovered without antibiotic therapy or further surgery, but 59 (29.5%) developed complications and 10 (5%) died. The commonest complication was abscess formation, and the incidence was highest with infections associated with malignancy (44%) and lowest with obstetric infections (5%). The mortality was 5% overall but in the presence of bacteraemia it rose to 33%.Only 43 patients received appropriate chemotherapy. Clindamycin was the most effective antibiotic, having a recovery rate of 78%, but this rate was little better than in untreated patients (65%). The role of prophylactic antibiotic therapy in preventing bacteroides infection remains to be studied.The incidence of the isolation of bacteroides from wound infections after major intestinal surgery rose from 13% in 1970 to 81% in 1973. This increase was due to both the accurate collection and care of specimens while in transit to the laboratory and the use of selective media for the isolation of bacteroides in laboratory culture. The importance of these precautions is emphasized.

A strain of Myxococcus xanthus, referred to as M. xanthus TA, has been isolated which produces an antibiotic capable of inhibiting growth of certain Gram-positive and Gram-negative bacteria. Antibiotic production was significantly inhibited when the concentration of protein hydrolysate in the production medium exceeded 1%. The antibiotic was purified over 1000 times to apparent homogeneity by silicic acid chromatography and by a variety of preparative alumina thin-layer chromatographic procedures. The purified antibiotic was partially characterized by its chromatographic behavior in six solvent systems, stability to acid, alkali and heat, infrared, ultraviolet, and mass spectra. The antibiotic has a λmax of 242 nm in methanol.

Tobramycin and amikacin are new aminoglycoside antibiotics which resemble gentamicin and kanamycin respectively in pharmacokinetic, toxicological, and some microbiological respects. Minimum inhibitory concentrations for these four antibiotics have been determined against 393 bacterial strains (13 species from eight genera) isolated from clinical materialKanamycin was the least active, 89% of the Pseudomonas aeruginosa strains being resistant. Resistance was also found in 21% of the Klebsiella aerogenes, 16% of the indoleproducing Proteus species, 4% of the E. coli and 3% of the Enterobacter spp. tested. Providenca stuartii strains were often resistant to gentamicin and tobramycin.Apart from these exceptions, however, all the organisms tested were sensitive to all four compounds, gentamicin and tobramycin usually showing the highest intrinsic activity.Amikacin was the most active compound against a selection of strains chosen because of their resistance to one or more of the aminoglycosides. Thus, amikacin represents a significant advance over kanamycin and seems to be superior to gentamicin in terms of its in-vitro properties. This is especially the case in those unusual circumstances when gentamicin-resistant strains are a problem. We suggest that amikacin warrants further investigation since it may prove to be a valuable alternative to gentamicin in the treatment of life-threatening infections.

Within a period of only seven months three patients in one hospital under treatment with antibiotics by intravenous infusion developed secondary bloodstream infection originating from the site of insertion of the cannula. Two of these infections were fatal. Precautions suggested include strict asepsis, the skin being sterilized with iodine in spirit solution; antibiotic spray; and changing the cannula site.

Strains of Escherichia coli are resistant to the antibiotic kasugamycin due to the partial non-methylation of 16S ribosomal RNA. An RNA methylase activity, absent from resistant strains, is shown here to methylate in vitro the 16S RNA of resistant as well as sensitive strains.

R factors conferring a high level of resistance to trimethoprim have been identified in one strain of Escherichia coli and one of Klebsiella aerogenes, both isolated from infected urines.

The sideromycin antibiotic A 22,765 is taken up from the medium by Staphylococcus aureus SG 511 against a concentration gradient. In contrast to this, A 22,765-resistant cells display an extremely low degree of permeability to the molecule. Ferrioxamine B penetrates the cells in only very minute quantities, but nevertheless antagonises permeation by A 22,765. This permeation-antagonising effect of ferrioxamine B is discussed.

Kalafungin is a new antimicrobial agent obtained from the culture broth of a soil isolate of Streptomyces tanashiensis , designated Streptomyces tanashiensis strain Kala UC-5063. Kalafungin is a chemically stable, nonpolyene agent which is ex-extremely inhibitory in vitro against a variety of pathogenic fungi, yeasts, protozoa, gram-positive bacteria, and, to a lesser extent, gram-negative bacteria.

An actively antibiotic producing actinomycete, Streptomyces NRC-101, was isolated from Egyptian soil. The strong antibiotic effect of the fermentation broth of the microorganism on grampositive and gramnegative bacteria led to experiments for the isolation of the active principle. After isolation and purification its physico-chemical and biological properties were different from the well-known antibiotics. Therefore, it was identified as a new antibiotic designated as antibiotic NRC-101.

Three new antibiotics, neoantimycin, neaureothin and ochramycin have been isolated from Streptoverticillium orinoci. The systematic position of the organism is discussed. Neoantimycin is related to the antimycin antibiotics group; neoaureothin shows some similarity with aureothin. Ochramycin is an amphoteric compound containing in its molecule an octaene chromophore. The first two antibiotics show a slight antifungal activity only. Ochramycin is active against gram +ve, but not against gram -ve bacteria; its activity against yeasts and fungi is very slight. Toxicity data are reported; no therapeutical effectiveness has been observed in experimental infections in mice.

Acute epiglottitis is a fulminating infection of young children, requiring urgent and specific treatment. Most cases are caused by infection with Haemophilus influenzae type b. Only a few cases, nearly all of them fatal, have been reported from Britain, but probably many others go unrecognized. Of four cases seen by us in the past six years three survived, having responded rapidly and completely to treatment. This consisted chiefly of measures to deal with respiratory tract obstruction and parenteral administration of antibiotics. There is need for a widespread awareness of the existence, presenting features, and extreme urgency for treatment of this disease.

In a survey of adverse drug reactions in wards of two Belfast hospitals for 52 weeks in 1965–6, 2·9% of 1,268 patients seen were admitted to hospital because of adverse reactions to drugs taken for therapeutic reasons and 2·1% were admitted because of self-poisoning. Patients admitted because of adverse drug reactions were older than those admitted because of self-poisoning and stayed in hospital longer. Among the drugs which caused the adverse reactions were digitalis preparations, antibiotics, corticosteroids, anticoagulants, analgesics, and tranquillizers. Hypersensitivity and side-effect types of reactions were the most common. Barbiturates were the most frequently used drugs in suicidal attempts.

Ferrous sulphate administered together with tetracycline and three of its derivatives-oxytetracycline, methacycline, and doxycycline-was found seriously to impair the absorption of these antibiotics. Thus even small doses of iron taken simultaneously should be avoided during tetracycline treatment.

1. The urinary excretion of total sulphanilamide (free and acetylated) in rats receiving Prontosil or Neoprontosil orally is considerably reduced when the rats are treated with antibiotics to suppress their intestinal flora. The absorption and metabolism of sulphanilamide are unaffected by such treatment with antibiotics.2. The lipid-soluble Prontosil after oral or intraperitoneal administration is partly excreted in the bile as a polar conjugate, apparently an N-glucuronide, and this drug appears to be converted into sulphanilamide partly by metabolism by the body tissues and partly by enterofloral metabolism.3. The polar, water-soluble Neoprontosil is poorly absorbed from the intestine and when given orally is largely reduced to sulphanilamide by the gut flora before absorption. When given intraperitoneally, a large proportion (up to 70%) of the drug is excreted in the bile unchanged, and it would appear that only a small proportion of the drug (ca. 20%) is converted to sulphanilamide in the tissues, th...

A rise in Salmonella typhimurium infection was observed in calves in Britain during 1964–6, follwing the adoption of the intensive farming method. A single phage type of S. typhimurium, type 29, was incriminated as the major pathogen. Attempts to treat and control the disease with a range of antibiotics were ineffective, but resulted in the acquisition of transferable multiple drug resistance by type 29. The transmission of drug-resistant type 29, directly or indirectly, from bovines to man resulted in many human infections. Transferable drug resistance reaching man from enterobacteria of animal origin may ultimately enter specifically human pathogens. Infections such as that caused by type 29 can be eliminated, not by the massive use of antibiotics but by improvement in conditions of animal husbandry and reduction in the opportunities for the initiation and spread of the disease. A reappraisal is needed of the methods of using antibiotics to determine how these methods can be improved, in order to conserve the long-term efficacy of the antibiotics.

Five cases of coincident salmonellosis and ulcerative colitis are described. In three the diagnostic combination was recognized late. If corticosteroid therapy is given for the colitis once the combination is recognized, then simultaneous systemic antibiotic cover is advisable. One of the five patients died during corticosteroid therapy alone with salmonella septicaemia due to a usually non-invasive organism.