[Extraction and purification of Micrococcus pyogenes nucleodepolymerase].
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In addition to antibiotic therapy associated with immobilization in a plaster cast, effective in the majority of cases of acute arthritis caused by St. pyogenes and resection-arthrodesis which is necessary in the stage of osteoarthritis, synovectomy occupies an intermediate position. During a period--possibly not a short one--when the lesions are still purely synovial and do not regress under medical treatment, their ablation permits recovery from the infection with restoration of the functional activity of the joint. The experience is based on 26 observations (19 knees, 3 hips, 2 shoulders, 1 thumb, 1 metacarpo-phalangeal lesion). In 2/3 of cases these arthritides were iatrogenic (13 after cortisone infiltration, 4 postoperative, 1 haematogenic from i.v. catheterization). Synovectomy should be complete; haemostasis must be rigorous; an aspiration drain is left in place for 5 or 6 days; additional immobilization in plaster is not obligatory; antibiotic treatment is continued until the BSR has returned to normal. As regards control of the infection, this was obtained in 19 of 26 operations. As regards function, in 10 of these cases the results were very good, in 7 good (slight persistent restriction of movement), in only 2 cases the results were unsatisfactory. The factors that appear to affect the results are on the one hand rigorous technique and on the other sufficiently early operation before the lesions have become too fully established. However, a study of arthritides after cortisone infiltration of previously affected joints has shown that even relatively late operation can lead to satisfactory results (9 of 13).
1. RNA was synthesized in vitro from a template of bacteriophage T4 DNA, in the presence of Mn(2+). A comparison was made of the RNA synthesized by purified RNA polymerase from two sources, Micrococcus lysodeikticus and Escherichia coli; these are referred to as Micrococcus cRNA and E. coli cRNA respectively (where cRNA indicates RNA synthesized in vitro by using purified RNA polymerase and a DNA primer). 2. Both types of RNA were self-complementary as judged by resistance to digestion with ribonuclease after self-annealing, Micrococcus cRNA being more self-complementary (40%) than was E. coli cRNA (30%). The cRNA was found to be much less self-complementary if Mg(2+) was present during RNA synthesis instead of Mn(2+). 3. Micrococcus cRNA hybridized with a larger part of bacteriophage T4 DNA than did E. coli cRNA. The E. coli cRNA competed with only part (70%) of the Micrococcus cRNA in hybridization-competition experiments. It is concluded that more sequences of bacteriophage T4 DNA are transcribed by Micrococcus polymerase than by E. coli polymerase. 4. The RNA sequences synthesized by Micrococcus RNA polymerase but not by E. coli RNA polymerase are shown by hybridization competition to compete with specifically late bacteriophage T4 messenger RNA sequences. The relevance of this finding to the control of transcription is discussed. 5. In an Appendix, new methods are described for the analysis of hybridization-saturation and -competition experiments. Particular attention is paid to the effects produced if different RNA sequences are present at different relative concentrations. 6. By using cRNA isolated from an enzymically synthesized DNA-RNA hybrid, it is estimated that, of the DNA that is complementary to cRNA, only about half can become hybridized with cRNA under the experimental conditions used.
The organisms studied were those of the family Micrococcaceae which cannot participate in genetic exchange with Micrococcus luteus and those whose biochemical and physiological characteristics appear to bridge the genera Staphylococcus and Micrococcus. The hydrocarbon compositions of M. luteus ATCC 4698 and Micrococcus sp. ATCC 398 were shown to be similar to those previously reported for many M. luteus strains, consisting of isomers of branched monoolefins in the range C25 to C31. However, Micrococcus sp. ATCC 398 differed somewhat by having almost all C29 isomers (approximately 88% of the hydrocarbon composition). Micrococcus spp. ATCC 401 and ATCC 146 and M. roseus strains ATCC 412, ATCC 416, and ATCC 516 contained the same type of hydrocarbon patterns, but the predominant hydrocarbons were within a lower distribution range (C23 to C27), similar to Micrococcus sp. ATCC 533 previously reported. The chromatographic profile and carbon range of the hydrocarbons of an atypical strain designated M. candicans ATCC 8456 differed significantly from the hydrocarbon pattern presented above. The hydrocarbons were identified as branched and normal olefins in the range C16 to C22. Studies of several different strains of staphylococci revealed that these organisms do not contain readily detectable amounts of aliphatic hydrocarbons. The members of the family Micrococcaceae have been divided into two major groups based on the presence or absence of hydrocarbons. With the exception of M. candicans ATCC 8456, this division corresponded to the separation of these organisms according to their deoxyribonucleic acid compositions.
Studies were conducted to investigate the removal and recovery of copper (II) ions from aqueous solutions by Micrococcus sp., which was isolated from a local activated sludge process. The equilibrium of copper biosorption followed the Langmuir isotherm model very well with a maximum biosorption capacity (q(max)) of 36.5 mg of Cu2+/g of dry cell at pH 5.0 and 52.1 mg of Cu2+/g of dry cell at pH 6.0. Cells harvested at exponential growth phase and stationary phase showed similar biosorption characteristics for copper. Copper uptake by cells was negligible at pH 2.0 and then increased rapidly with increasing pH until 6.0. In multimetal systems, Micrococcus sp. exhibited a preferential biosorption order: Cu approximate to Pb > Ni approximate to Zn. There is virtually no interference with copper uptake by Micrococcus sp. from solutions bearing high concentrations of Cl-, SO4(2-), and NO3- (0-500 mg/L). Sulfuric acid (0.05 M) was the most efficient desorption medium, recovering >90% of the initial copper sorbed. The copper capacity of Micrococcus sp. remained unchanged after five successive sorption and desorption cycles. Immobilization of Micrococcus sp. in 2% calcium alginate and 10% polyacrylamide gel beads increased copper uptake by 61%. Biomass of Micrococcus sp. may be applicable to the development of potentially cost-effective biosorbent for removing and recovering copper from effluents.
1. In a previous study, the differing sensitivity of bacterial strains as they occur in nature appeared to be correlated with their correspondingly differing reactivity with penicillin. Presumably, the over-all reactivity of the cell with penicillin paralleled that of the vulnerable cell component(s). However, when penicillin-resistant variants of these strains (Streptococcus pyogenes, Micrococcus pyogenes, Diplococcus pneumoniae, and Streptococcus faecalis) were produced by serial passage through increasing concentrations of antibiotic, this correlation between resistance and the ability of the cell to bind penicillin was no longer apparent. Some resistant variants bound more penicillin than the parent, sensitive cell (Streptococcus faecalis, Micrococcus pyogenes), some were unchanged in their reactivity (Diplococcus pneumoniae, Micrococcus pyogenes), and some bound less (Streptococcus pyogenes, Micrococcus pyogenes). One resistant variant of Micrococcus pyogenes at first showed enhanced reactivity with penicillin; on continued passage through antibiotic, there was a further increase in resistance, but now associated with a significantly decreased reactivity. In the case of Diplococcus pneumoniae, a resistant variant at first reacted normally with penicillin; on continued passage in antibiotic, its binding affinity for penicillin gradually decreased, but with no associated further increase in resistance. 2. The reactivity with penicillin of cell-free sonic extracts of the resistant variants paralleled that of the intact organisms. Permeability considerations therefore did not seem involved in the increased resistance produced by serial passage in antibiotic. 3. The penicillin-resistant variants did not have an enhanced capacity to degrade the free intracellular antibiotic. 4. Possible alternative explanations are discussed in the text.
In a comparative study 351 strains of Micrococcus subgroups 1-3 (Baird-Parker, 1963, 1965, 1974) were classified according to the Kloos and Schleifer scheme (1975b). The results showed that Micrococcus subgroups 1, 2 and 3 are heterogeneous groups in the Kloos and Schleifer scheme. Strains belonging to Micrococcus subgroups 1 and 2 were mostly classified by the Kloos and Schleifer criteria as Staphylococcus hominis, Micrococcus subgroup 3 strains from the skin as S. cohnii, while Micrococcus subgroup 3 strains from urinary infections were classified mainly as S. saprophyticus. The correlation of novobiocin resistant S. saprophyticus biotype III (Baird-Parker, 1974) with S. saprophyticus (Kloos and Schleifer, 1975a, 1975b) when isolated from urine, was 80%. Although the Kloos and Schleifer scheme provides more information about biochemical characters, doubts are expressed about the validity of some of the species so delineated.
A study of the bacterial flora of the genitalia of nulliparous Savanna Brown does was carried out both before breeding and at different intervals postpartum to investigate the type of microbial organisms that could be present in the uterus, cervix and the vagina respectively. Of 29 pre-breeding vaginal swabs, Staphylococcus sp. was isolated from 20 goats, Streptococcus sp. from 15 goats and Micrococcus sp. from four goats. Mycoplasma agalactiae was isolated from five goats. The postpartum vagina did not show any appreciable change in the microbial flora, except that Escherichia coli was encountered in two cases. The uterus yielded E. coli from the goats slaughtered 2 days postpartum; Micrococcus sp. from goats slaughtered 12 days postpartum; Staph. aureus from goats slaughtered 16 days postpartum and Staph. aureus from goats slaughtered 24 days postpartum. Also in these two cases-a goat slaughtered at two days postpartum (dpp) and a goat slaughtered 16 dpp-E. coli was present in the uterus. Other isolates from the uteri of slaughtered goats were Micrococcus sp. (12 dpp), Staph. aureus and Micrococcus sp. (16 dpp) and Staph. aureus (24 dpp).
The hemolymphs of two diplopod (Chicobolus sp. and Rhapidostreptus virgator) and two chilopod species (Lithobius forficatus and Scolopendra cingulata) were tested for the presence of antibacterial substances using Petri dish tests. The native hemolymph of all species had substances acting on living Micrococcus luteus, whereas only Rhapidostreptus, Scolopendra, and Lithobius were effective against lyophilized Micrococcus. The antibacterial activity against living Micrococcus increased after inoculation with bacteria (Enterobacter cloacae beta-12) in Chicobolus and Rhapidostreptus and also against lyophilized Micrococcus in the latter. Thus, these effects appear to be inducible. None of the myriapods tested had any bacteriostatic effect on Escherichia coli D-31 whereas the growth of gram-negative E. cloacae was inhibited. The antibacterial substances in the diplopod species were unstable when heated but were resistant to freezing. At least two antibacterial substances (a lysozyme-like one and another substance) are considered to occur in Myriapoda.
Defined media permitting extensive growth of representative pink radio-resistant tetracocci (Micrococcus radiodurans, Micrococcus roseus, and Micrococcus radiophilus) and two controls (an ultraviolet-sensitive mutant of M. radiodurans and Micrococcus luteus) are described. Availability of Fe (especially Fe3+) proved essential for good growth, as evidenced by (i) favorable effects of hydroxamic acids, e.g., salicylhydroxamic acid, and (ii) the growth promotion by hemin when joined with elevated concentrations of Fe. Cobalamin (B12) and methionine were interchangeable as an absolute requirement for methionine not affected by B12. M. luteus required neither. Pink radio-resistant micrococci may form a coherent group. Some divergences among them might be attributable to the method for isolating them, which for ordinary bacteria would be mutagenic to the point of total lethality. The ecology of these tetracocci vis-à-vis other pink-red radio-resistant organisms is discussed in relation to a question: can these bacteria be isolated without dependence on radiation as the cardinal selective factor?
IR spectra of thw whole cells of the coccoid forms (Mycococcus and Micrococcus) isolated from lithophilous lichen were compared with IR spectra of the collection cultures of Micrococcus and Arthrobacter. Generic spectral characteristics of Mycococcus and Micrococcus are presented. Spectral heterogeneity within the genus Arthrobacter complicates the diagnosis. The cultures of the Mycococcus genus were divided into three groups according to their spectral characteristics. Spectral scans of the studied coccoid forms differ from the scans of the mycelial actinomycetes, and their intensities within the range of 900-1200 cm-1 (lipids) decrease in the series Mycococcus, Arthrobacter, Micrococcus.
Antibodies induced by the gram+ bacteria Micrococcus lysodeikticus exhibit different carbohydrate specificities and hence might cross-react with membrane glycoproteins and/or glycolipids of mammalian cells. Using a T cell derived lymphoid line, these antibodies were found to detect a membrane marker which is only exposed in confluent culture conditions on non-dividing cells. Such confluence related antigen (Cag) is a cryptic membrane antigen, which can be unmasked through membrane perturbating agents such as p-formaldehyde or through interactions with macrophages and macrophage derived factors. Anti-micrococcus antibodies appear also to affect functionally normal T lymphocytes. Thus such reagents drastically inhibit the murine T cell reactivity towards mitogens such as Con A and PHA provided, however, the mitogenic signal is delivered through peritoneal macrophages. Furthermore, anti-micrococcus antibodies induce activated T lymphocytes into mitogenesis, but not unprimed resting T cells. Hence the physiological activity of anti-micrococcus antibodies depends on the state of activation of the T lymphocyte, indicating the involvement of cryptic membrane molecules. The relevance of such phenomena to host-bacteria and host-parasite interaction will be discussed.