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PubMed1992

In vitro hydrogen production by enteric bacteria cultured from children with small bowel bacterial overgrowth.

Lactulose breath hydrogen test and Enterotest string test were carried out simultaneously on 19 children 3-5 years old. Bacteria isolated from the jejunal fluid in upper small intestines of these children were incubated with lactulose at neutral pH. Anaerobes were present in all but one child, and in 15 children they were present in numbers greater than 5 log10 organisms per ml. Most of these bacteria did not produce hydrogen in vitro. Hydrogen production (greater than 100 ppm) was observed with the following bacteria: Bacteroides (5%), clostridia (8%), anaerobic corynebacteria (5%), Escherichia coli (67%), Lactobacillus (8%), Staphylococcus (8%), and Streptococcus (9%). The following bacteria did not produce hydrogen in vitro: Actinobacter, Actinomyces, anaerobic cocci, Bifidobacterium, Fusobacterium, micrococci, Neisseria, Sarcina, and Veillonella. This study suggests that in the diagnosis of small bowel bacterial overgrowth using lactulose breath hydrogen test, it is important to consider that patients with a flat breath hydrogen response to a carbohydrate challenge during the first 60 min may be infected with enteric bacteria which are not capable of producing H2.

Bacteria↗

PubMed2001

Methylosarcina fibrata gen. nov., sp. nov. and Methylosarcina quisquiliarum sp.nov., novel type 1 methanotrophs.

Two novel species of obligate methane-oxidizing bacteria, isolated from landfill soil, were characterized. Both strains were unusual in that some members of the population grew in irregularly shaped, refractile cell packets that resembled sarcina-like clusters. Electron microscopy revealed that the cell packets were covered with a slime layer and the cells contained many large granular inclusion bodies. The individual cells of each strain were sometimes motile and had differing morphologies. Isolate AML-C10T was always coccoidal in shape, and the cells were covered with extracellular fibrils. Isolate AML-D4T was pleomorphic, changing from rod to coccal form, sometimes exhibiting an unusual fusiform morphology. AML-D4T lacked the extensive fibrillar matrix observed with AML-C10T. Both strains utilized only methane and methanol as carbon sources. In stationary phase, the cells of each strain swelled in size and formed cysts. Aside from morphological differences, strains could also be distinguished from each other by cellular protein patterns, as well as by temperature and pH tolerances. 16S rDNA phylogenetic analysis showed that these are type I methanotrophs (family: Methylococcaceae) most closely related to the Methylobacter/Methylomicrobium clade, although they form a monophyletic grouping supported by moderately high bootstrap values. By 16S rDNA database searches, the most similar species to both isolates were Methylobacter spp. However, partial particulate methane monooxygenase sequence analysis suggested that these bacteria might be more closely related to Methylomicrobium than Methylobacter. Furthermore, cellular fatty acid profiles of the strains more closely resemble those of Methylomicrobium, although the absence of significant levels of 16:1omega5c argues for the uniqueness of these two strains. On the basis of the results described here, it is proposed that a new genus should be created, Methylosarcina gen. nov., harbouring two species, Methylosarcina fibrata sp. nov. (type species) and Methylosarcina quisquiliarum sp. nov. The type strains are AML-C10T (= ATCC 700909T = DSM 13736T) and AML-D4T (= ATCC 700908T = DSM 13737T), respectively.

Bacterial Proteins↗

PubMed1977

Excystment of axenically prepared cysts of Hartmanella culbertsoni.

Axenically prepared cysts of Hartmannella culbertsoni readily excysted in the presence of heat stable factors prepared from Escherichia coli, Klebsiella aerogenes, Staphylococcus aureus, Sarcina lutea, Bacillus subtilis, Bacillus megaterium and several fungi. Peptone, proteose peptone, tryptone or amino acids also promoted excystment. Crowding of the cysts and dilution of bacterial extracts adversely affected the excystment. Continual presence of the factors in the medium was essential for excystment.

Amino Acids↗

PubMed1994

Lautropia mirabilis gen. nov., sp. nov., a gram-negative motile coccus with unusual morphology isolated from the human mouth.

An organism that seems to be identical to Orskov's 'Sarcina mirabilis' [Orskov, J. (1930) Acta Pathol Microbiol Scand Suppl III, 519-541] has been rediscovered in specimens from the upper respiratory tract of humans. Six strains were studied, and the results, which conformed to Orskov's description of S. mirabilis, were as follows. Rough to smooth colonies grow on many plated media and show extremely polymorphic cell morphology with round cells with diameters from 1 to > 10 microns. The smallest cells were often motile with circular movements. Strains were Gram-negative, facultatively anaerobic, oxidase and urease positive, and weakly catalase positive. Nitrate and nitrite were reduced, and glucose, fructose, sucrose and mannitol were fermented. Polysaccharide was produced on sucrose agar. Electron microscopy showed coccoid cells with a bundle of three to nine flagella, a Gram-negative cell-wall morphology, and aggregates of irregular cells held together by a common surface layer. The mean mol% (G+C) of the organisms was 65.0. 16S-ribosomal RNA sequencing revealed that the organism belongs to the beta subgroup of Proteobacteria, separate from all other described genera, but most closely related to Burkholderia. The name Lautropia mirabilis is proposed for this organism.

Bacterial Typing Techniques↗

PubMed2005

Construction and expression of an ethanol production operon in Gram-positive bacteria.

Pyruvate decarboxylase (PDC), an enzyme central to homoethanol fermentation, catalyses the non-oxidative decarboxylation of pyruvate to acetaldehyde with release of carbon dioxide. PDC enzymes from diverse organisms have different kinetic properties, thermal stability and codon usage that are likely to offer unique advantages for the development of desirable Gram-positive biocatalysts for use in the ethanol industry. To examine this further, pdc genes from bacteria to yeast were expressed in the Gram-positive host Bacillus megaterium. The PDC activity and protein levels were determined for each strain. In addition, the levels of pdc-specific mRNA transcripts and stability of recombinant proteins were assessed. From this analysis, the pdc gene of Gram-positive Sarcina ventriculi was found to be the most advantageous for engineering high-level synthesis of PDC in a Gram-positive host. This gene was thus selected for transcriptional coupling to the alcohol dehydrogenase gene (adh) of Geobacillus stearothermophilus. The resulting Gram-positive ethanol production operon was expressed at high levels in B. megaterium. Extracts from this recombinant were shown to catalyse the production of ethanol from pyruvate.

Ethanol↗

PubMed1978

Plasma, bone, hip capsule and drain fluid concentrations of ampicillin and flucloxacillin during total hip replacement after intravenous bolus injection of magnapen.

1. A rapid intravenous bolus injection of 4.0 g Magnapen (which contains 2.0 g of ampicillin and 2.0 g of flucloxacillin) was to seven patients undergoing total hip replacement immediatly before induction of general anesthesia. Postoperatively the patients patients received 2.0 g Magnapen by intramuscular injection every 6 h for up to 72 h until removal of the wound drains. 2. The plasma, bone, hip capsule and drain fluid concentrations of ampicillin and flucloxacillin were measured by a differential small plate microbiological assay method using Sarcina lutea and a penicillinase producing Staph. aureus Russell as the test organisms. 3. The mean +/- s.e. mean concentrations of ampicillin after this regimen were 4222.2 +/- 285.0 microgram/ml (plasma), 65.6 +/- 1.3 microgram (g (hip capsule), 19.1 +/- 3.8 microgram/g (cancellous bone), and 211.1 +/- 65.6 microgram/g (ground up bone) respectively. 4. The mean +/- s.e. mean flucloxacillin concentrations after this regime were 137.2 +/- 28.4 microgram/ml (plasma), 61.8 +/- 15.0 microgram/g (hip capsule), 47.1 +/- 9.5 microgram/g (cancellous bone) and 139.4 +/- 21.8 microgram/g (ground up bone) respectively. 5. An intravenous bolus injection of Magnepen (4.0 g), given immediately before induction of general anaesthesia, provides concentrations of ampicillin and flucloxacillin in plasma, hip capsule, cancellous and ground up bone, and drain fluid that exceed the MICs of these antibiotics against Staph. aureus and E. coli. 6. The plasma, hip capsule, cancellous and ground up bone concentrations of ampicillin after this dose of Magnapen do not, however, exceed the MICs of the Gram negative anaerobes that sometimes cause postoperative wound infections in these patients.

Aged↗

PubMed1984

A note on the temperature tolerance of Legionella.

A strain of Legionella pneumophila serogroup 1 isolated from the environment had a decimal reduction time in water at 50 degrees C (D50) of 111 min, a D54 of 27 min and a D58 of 6 min. There was little loss of viability at 46 degrees C. Other environmental organisms, a Pseudomonas sp., a Micrococcus sp. and a coliform survived less well at these temperatures. A species of Sarcina had a survival time greater than the L. pneumophila at all the temperatures tested. Other strains of legionellas were tested at 50 degrees C and decimal reduction times calculated. These ranged from 80 min for another strain of L. pneumophia serogroup 1 to 216 min for L. bozemannii . Legionella micdadei did not survive well at 50 degrees C.

Legionella↗

PubMed1985

A note on the contamination of eye-drops following use by hospital out-patients.

Examination of 273 eye-drops, returned to a hospital pharmacy by out-patients showed that 27% were contaminated. Common contaminants included Bacillus spp, Staphylococcus albus and Sarcina spp. Those eye-drops containing the preservatives chlorbutol or thiomersal showed above average levels of contamination, thus questioning the suitability of these preservatives for ophthalmic preparations.

Bacteria↗

PubMedDate not supplied

Cephalexin concentration in the human ejaculate following oral and parenteral administration.

Cephalexin levels in the ejaculates of 31 infection-free volunteers have been determined following the administration of 1 g of antibiotic. In 16 subjects the cephalexin was administered by intramuscular injection and in 15 by mouth. In each case the volunteers were divided into groups of five each of which received the drug, 1, 2 or 4 hours before the collection of semen. Antibiotic concentrations were determined microbiologically on agar plates using Sarcina lutea ATCC 9341. For selection purposes healthy subjects were considered to be those showing a normal anamnesis and physical exploration, an absence of leucocytes demonstrable by a cytomorphological evaluation of the semen, semen citric acid and fructose levels within normal limits and a qualitatively and quantitatively similar saprophytic flora to that seen in a culture of urine taken immediately before the collection of semen. Maximum concentrations were obtained 2 hours after the administration of the antibiotic and the concentration following intramuscular injection (6.9 microgram/ml) was significantly greater than that following oral administration (4.7 microgram/ml).

Administration, Oral↗

PubMed1963

FURTHER PHARMACOLOGY AND CHEMOTHERAPY OF CLOXACILLIN.

In concentrations greatly in excess of therapeutic blood levels cloxacillin has a slight hypotensive action. As with other antibiotics, it sometimes causes diarrhoea in rabbits but, in doses up to 200 mg/kg, it does not have a teratogenic effect on the rabbit foetus. Cloxacillin is distributed throughout the body. High concentrations are found only in the liver and kidney, these reflecting the high concentrations in the bile and urine respectively. It differs from other penicillins investigated in that there appears to be little or no renal tubular secretion as demonstrated in experiments on the hen. Cloxacillin is excreted as the unchanged drug and as an active metabolite in the urine and bile. After oral administration it is metabolized in the caecum giving a penicillin which differs from the urinary metabolite and which has a greater antibiotic action against Sarcina lutea than the parent penicillin. The activity of the caecal metabolite against resistant and sensitive strains of Staphylococcus, however, remains similar to that of cloxacillin. Against infections due to resistant and sensitive Staphylococcus in animals cloxacillin is active by both the oral and subcutaneous routes and it is more effective orally than an equal subcutaneous dose of methicillin.

Animals↗

PubMed1963

Antimicrobial substances from aspen tissue grown in vitro.

Isolated aspen tissue, when grown in vitro for 3 weeks on agar medium, yielded antimicrobial substances which produced inhibitory zones when the culture plates were inoculated with Fusarium roseum, Saccharomyces cervisiae, Bacillus subtilis, Bacillus cereus, Penicillium, roqueforti, Torula utilis, Sarcina lutea, Flavobacterium aquatile, Pullularia pullulans, and Staphylococcus aureus.

Anti-Infective Agents↗

PubMed1977

Chartreusin: production and microbiological assay.

Chartreusin was produced in the fermentation liquors of Streptomyces chartreusis at peak concentrations of 200 to 300 mug/ml. The titers could be increased by 200 to 300% or more by incorporating d-fucose, a part of the chartreusin molecule, into the fermentation media. A microbiological assay with Sarcina lutea could detect concentrations of the drug of 0.5 to 1.0 mug/ml.

Antibiotics, Antineoplastic↗

PubMed1980

Bioassay of antibiotics in body fluids from patients receiving cancer chemotherapeutic agents.

Patients receiving antitumor chemotherapy are at increased risk of developing nosocomial infections, and the antibacterial therapy of such infections is often monitored by bioassay. The effect of antitumor agents on seven bioassay procedures using strains of Sarcina, Klebsiella, Clostridium, Pseudomonas, Staphylococcus aureus, and S. epidermidis or Bacillus was evaluated. The minimum inhibitory concentrations of six antitumor drugs, cytarabine, dactinomycin, doxorubicin, 5-fluorouracil, methotrexate, and vinblastine, determined for each of the test organisms, showed that 5-fluorouracil, dactinomycin, and doxorubicin are used at blood levels sufficient to interfere with bioassay procedures. The other drugs have minimum inhibitory concentrations as much as 100 times the expected blood levels. Antibiotic (gentamicin, kanamycin, cephalothin, and carbenicillin) recovery experiments in the presence of therapeutic levels of antitumor agents showed no in vitro inactivation of antibiotic. However, at low cephalothin concentrations (less than 20 microgram/ml) in the presnce of 5-fluorouracil, bioassay results were in error by as much as 100%. The data indicate that bioassay procedures for the determining of antibacterial drug levels may need to be modified for those patients receiving antitumor therapy with 5-fluorouracil, doxorubicin, or dactinomycin.

Anti-Bacterial Agents↗

PubMed1972

Activity of methacycline, related tetracyclines, and other antibiotics against various L-forms and their parent bacteria in vitro.

The activity of methacycline against microbial L-forms and their parent bacteria was compared with that of oxytetracycline, chlortetracycline, tetracycline, and demethylchlortetracycline, as well as with that of 22 other antibiotics which included examples of major groups of antibiotics. The L-forms and bacteria used were Streptococcus faecalis, S. faecium, S. faecalis var. zymogenes, Staphylococcus aureus (three strains), Proteus mirabilis (two strains), Pseudomonas aeruginosa, Escherichia coli (two strains), Sarcina flava, Serratia marcescens, and Klebsiella pneumoniae. The five tetracyclines had similar activities and were more active against L-forms than bacterial forms, except that the bacterial form of S. flava was more susceptible than the L-form. In general, other antibiotics (except the penicillins) were more active against L-forms than bacterial forms. There were certain exceptions where the bacterial form was more susceptible than the L-form. These included the effect of polymyxin B and colistin on P. aeruginosa, E. coli, and P. mirabilis, and the effect of gentamicin on P. aeruginosa, E. coli, S. flava, and S. marcescens.

Anti-Bacterial Agents↗

PubMed1986

Pharmacokinetics of cefotaxime, moxalactam, and cefoperazone in the early puerperium.

Twelve parturient women volunteered to receive 1 g of cefotaxime on the second or third day postpartum by intravenous infusion over 3 min. Blood was taken from the antecubital vein of the contralateral arm at 0.25, 0.5, 0.75, 1, 2, 4, and 6 h. The concentration of cefotaxime in serum was assayed by agar diffusion, with Sarcina lutea ATCC 9341 as the indicator strain. The same 12 women received an identical dose of antibiotic 4 months after the first dose, and blood was taken at the same time intervals as in the first study to measure antibiotic levels. An additional 24 women participated in identical studies with either moxalactam or cefoperazone. Cefoperazone afforded the highest concentration in serum of the three drugs, followed by moxalactam. These differences in the concentration in serum were seen both early postpartum and 4 months later. However, the concentration in serum of all three drugs was diminished 2 and 3 days postpartum compared with 4 months postpartum. Most pharmacokinetic parameters were also significantly altered early in the puerperium relative to those obtained 4 months later. The altered pharmacokinetic behavior of antibiotics associated with pregnancy appears to persist into the early puerperium.

Adult↗

PubMed1977

Microbial conversion of DL-2-amino-delta2-thiazoline-4-carboxylic acid to L-cysteine and L-cystine: screening of microorganisms and identification of products.

Microorganisms able to form L-cysteine from DL-2-amino-delta2-thiazoline-4-carboxylic acid (DL-ATC), a chemical intermediate in the synthesis of DL-cysteine, were isolated from soil samples and classified as Pseudomonas sp., Pseudomonas cohaerens, P. desmolytica, and P. ovalis. Thirteen L-cysteine-producing bacteria were also found in among 463 stock cultures representing 37 genera. These were Achromobacter delmarvae. Alcaligenes denitrificans, Bacillus brevis, Brevibacterium flavum, Enterobacter aerogenes, Erwinia carotovora, Escherichia coli, Micrococcus sodonensis, Myocoplana dimorpha, Sarcina lutea, Serratia marcescens, Flavobacterium acidoficum, and Pseudomonas ovalis. In the presence of intact cells of Pseudomonas sp. AJ 3854, 6.1 mg of L-cysteine and/or L-cystine per ml was produced from 10 mg of DL-ATC-3H2O per ml in a molar yield of 100%. This finding suggests that racemization and asymmetric hydrolysis occurred simultaneously in this incubation mixture. After the complete oxidation of cysteine to cystine by aeration in the presence of ferrous ion, crystalline cystine was isolated; its configuration was the L isomer based on data from X-ray diffraction, microbioassay, and optical rotation studies.

Bacteria↗

PubMed1983

Methanogenic bacteria from the bondyuzhskoe oil field: general characterization and analysis of stable-carbon isotopic fractionation.

Selective enrichment culture techniques were employed to obtain mixed cultures of methanogenic rods and sarcina from surface flooding waters and deep subsurface ( approximately 1650 m) oil-bearing sedimentary rocks and formation waters sampled from an old oil field in the U.S.S.R. previously reported to display active biological methanogenesis. The methanogens were selectively isolated as colonies on agar petri dishes that were incubated in a novel container. The general cellular and growth features of three Methanobacterium isolates were determined. These strains grew optimally at 37 to 45 degrees C in anaerobic pressure tube cultures with a doubling time of 16 to 18 h on H(2)-CO(2) and proliferated as autotrophs. Acetate addition significantly enhanced the final cell yield. Growth of these strains was completely inhibited by either 0.6 g of sodium sulfide per liter or 31.0 of sodium chloride per liter, but growth was not inhibited by either 0.3 g of sodium sulfide per liter or 1.0 g of sodium sulfate per liter. One novel isolate, Methanobacterium sp. strain ivanov, was grown on H(2)-CO(2), and the stable-carbon isotopic fractionations that occurred during synthesis of methane, cell carbon, and lipids were determined. The results of this study were used to examine the anomalous relationship between the isotopic and chemical compositions of natural gas occurring in the deep subsurface environment of the oil field.

Journal Article↗

PubMed1985

Nutrition and Growth Characteristics of Trichomitopsis termopsidis, a Cellulolytic Protozoan from Termites.

Putatively axenic cultures of Trichomitopsis termopsidis 6057, isolated by M. A. Yamin (J. Protozool. 25:535-538, 1978) from the hindgut of Zootermopsis termites, apparently contained methanogenic bacteria, inasmuch as small amounts of CH(4) were produced during growth. However, T. termopsidis could be "cured" of methanogenic activity by incubation in the presence of bromoethanesulfonate. Both the cured derivative (6057C) and the parent strain (6057) required NaHCO(3) and fetal bovine serum for good growth; the presence of yeast extract in media was stimulatory. Growth of both strains was markedly improved by substituting heat-killed cells of Bacteroides sp. strain JW20 (a termite gut isolate) for heat-killed rumen bacteria in media as a source of bacterial cell material. Heat-killed Bacteroides sp. strain JW20 was the best of a number of bacteria tested, and under these conditions H(2) was a major protozoan fermentation product. Growth of T. termopsidis strains was further improved by co-cultivation in the presence of Methanospirillum hungatii. M. hungatii was the best of a number of H(2)-consuming bacteria tested, and under these conditions CH(4), but not H(2), was produced, indicating interspecies transfer of H(2) between the protozoa and M. hungatii. Both strains of T. termopsidis used powdered, particulate forms of cellulose (e.g., pure cellulose, corncob, cereal leaves) as fermentable energy sources, although powdered wood, chitin, or xylan supported little or no growth. Cells of the cellulose-forming coccus Sarcina ventriculi also served as a fermentable energy source, but these were used poorly as a source of bacterial cell material. The only substantial difference between T. termopsidis 6057 and 6057C was that the latter grew poorly or not at all with rumen bacteria as a source of bacterial cell material. The improved growth of T. termopsidis in vitro should facilitate further studies on the cell biology and biochemistry of these symbiotic, anaerobic protozoa.

Journal Article↗