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Laboratory and field studies on the effects of the antibiotic tylosin on honey bee Apis mellifera L. (Hymenoptera: Apidae) development and prevention of American foulbrood disease.

Laboratory and field studies were conducted to determine the effectiveness of the antibiotic tylosin in preventing and controlling infections of American foulbrood disease (AFB) of honey bees. Studies conducted on immature worker bees maintained in the laboratory revealed that honey bee larvae could tolerate quite a range of doses of antibiotic in their diet. Intermediate doses of tylosin protected very young larvae from becoming infected by Bacillus larvae at a concentration of 1.5 x 10(8) spores/ml of diet. Antibiotic treatment had no measurable effects on larval or pupal developmental rates until the dose reached a lethal level. Bees in field colonies readily consumed tylosin in powered sugar, up to a level of 800 mg/7 g sugar. No negative colony effects were noted at any dosage rates. Protection against infection by American foulbrood was compared to results obtained with 200 mg Terramycin, the standard dose of the only substance currently registered for foulbrood control. Both 200 mg Terramycin and 100 mg tylosin protected the colonies for up to 3 weeks. A 200-mg dose of tylosin protected the colony for an additional week. Doses of 100 mg or more of tylosin were adequate to eliminate signs of AFB infection in overtly diseased colonies.

Animals↗

Characterization of isolates of Paenibacillus larvae subsp. larvae from diverse geographical origin by the polymerase chain reaction and BOX primers.

Ninety-nine strains of Paenibacillus larvae subsp. larvae, the causal agent of American Foulbrood disease (AFB) of honeybees, were isolated from different regions of Argentina and other countries. The isolates were characterized on the basis of DNA fingerprints by a polymerase chain reaction technique (PCR) with BOX sequence-specific primers. Isolates from Argentina generated three groups of patterns (designated A, B, and C), while P. larvae subsp. larvae strains obtained from other countries yielded two distinguishable patterns (coincident with A and B). Strains from U.S. A. and Germany were identical and related to Group A, while all Czech and English isolates belonged to Group B. Strains from France, Poland, Italy, Sweden, and New Zealand showed two different patterns (A and B). Comparisons of the biochemical type and genotype of isolates rendered no obvious linkage between both features. These results suggest that AFB in Argentina resulted from multiple sources of contaminated material.

Animals↗

The structure of the membrane protein squalene-hopene cyclase at 2.0 A resolution.

Squalene cyclases catalyze a cationic cyclization cascade, which is homologous to a key step in cholesterol biosynthesis. The structure of the enzyme from Alicyclobacillus acidocaldarius has been determined in a new crystal form at 2.0 A resolution (1 A=0.1 nm) and refined to an R-factor of 15.3 % (Rfree=18.7 %). The structure indicates how the initial protonation and the final deprotonation of squalene occur and how the transient carbocations are stabilized. The pathways of the flexible educt squalene from the membrane interior to the active center cavity and of the rigid fused-ring product hopene in the reverse direction are discussed. The enzyme contains eight so-called QW-sequence repeats that fortify the alpha/alpha-barrels by an intricate interaction network. They are unique to the known triterpene cyclases and are presumed to shield these enzymes against the released enthalpy of the highly exergonic catalyzed reaction. The enzyme is a monotopic membrane protein, the membrane-binding interactions of which are described and compared with those of two prostaglandin-H2 synthase isoenzymes, the only other structurally characterized proteins of this type. In the crystals the membrane-binding regions face each other, suggesting a micelle-type detergent structure between them.

Amino Acid Sequence↗

Stator structure and subunit composition of the V(1)/V(0) Na(+)-ATPase of the thermophilic bacterium Caloramator fervidus.

The V-type Na(+)-ATPase of the thermophilic, anaerobic bacterium Caloramator fervidus was purified to homogeneity. The subunit compositions of the catalytic V(1) and membrane-embedded V(0) parts were determined and the structure of the enzyme complex was studied by electron microscopy. The V(1) headpiece consists of seven subunits present in one to three copies, and the V(0) part of two subunits in a ratio of 5:2. An analysis of over 7500 single particle images obtained by electron microscopy of the purified V(1)V(0) enzyme complex revealed that the stalk region, connecting the V(1) and V(0) parts, contains two peripheral stalks in addition to a central stalk. One of the two is connected to the V(0) part, while the other is connected to the first via a bar-like structure that is positioned just above V(0), parallel with the plane of the membrane. In projection, this bar seems to contact the central stalk. The data show that the stator structure that prevents rotation of the static part of V(0) relative to V(1) in the rotary catalysis mechanism of energy coupling in ATPases/ATPsynthases is more complex than previously thought.

Adenosine Triphosphatases↗

Utilization of methoxylated benzoates and formation of intermediates by Desulfotomaculum thermobenzoicum in the presence or absence of sulfate.

Desulfotomaculum thermobenzoicum strain TSB (DSM 6193) was found to utilize some methoxylated benzoates as carbon and energy source with or without sulfate. 3- or 4-Methoxybenzoate, vanillate (4-hydroxy-3-methoxybenzoate), syringate (3,5-dimethoxy-4-hydroxybenzoate) and 3,4,5-trimethoxybenzoate were converted to corresponding hydroxybenzoates. However, neither 2-methoxybenzoate nor 2,6-dimethoxybenzoate was utilized. The organism grew acetogenically on each of the methoxylated benzoates in the absence of sulfate. 3,4-Dihydroxy-5-methoxybenzoate was detected during conversion of syringate, and syringate and 3,4-dihydroxy-5-methoxybenzoate were detected during conversion of 3,4,5-trimethoxybenzoate as intermediates. These findings indicate that 4-methoxyl-group is most readily cleaved, whereas 2-methoxyl-group is not utilized by the organism.

Acetates↗

Isolation and characterization of a new spore-forming sulfate-reducing bacterium growing by complete oxidation of catechol.

A new mesophilic sulfate-reducing bacterium, strain Groll, was isolated from a benzoate enrichment culture inoculated with black mud from a freshwater ditch. The isolate was a spore-forming, rod-shaped, motile, gram-positive bacterium. This isolate was able of complete oxidation of several aromatic compounds including phenol, catechol, benzoate, p- and m-cresol, benzyl alcohol and vanillate. With hydrogen and carbon dioxide, formate or O-methylated aromatic compounds, autotrophic growth during sulfate reduction or homoacetogenesis was demonstrated. Lactate was not used as a substrate. SO4(2-), SO3(2-), and S2O3(2-) were utilized as electron acceptors. Although strain Groll originated from a freshwater habitat, salt concentrations of up to 30 g.l-1 were tolerated. The optimum temperature for growth was 35-37 degrees C. The G + C content of DNA was 42.1 mol%. This isolate is described as a new species of the genus Desulfotomaculum.

Bacillaceae↗

Characterization of urease from Sporosarcina ureae.

Alkaline stable (pH 7.75-12.5) urease from Sporosarcina ureae was purified over 400-fold by ion exchange and hydrophobic interaction chromatography. The cytoplasmic enzyme was remarkably active with a specific activity of greater than 9300 mumol urea degraded min-1 mg protein-1 at pH 7.5, where it has optimal activity. Although S. ureae is closely related to Bacillus pasteurii, known to possess a homopolymeric urease containing 1 nickel per subunit [M(r) = 65000], the S. ureae enzyme is comprised of three subunits [apparent M(r) = 63,100 (alpha), 14,500 (beta), and 8500 (gamma)] in an estimated alpha beta gamma 2 stoichiometry and contains 2.1 +/- 0.6 nickel ions per alpha beta gamma 2 unit as measured by atomic absorption spectrometry. Stationary phase cultures sometimes possessed low levels of urease activity, but the specific activity of cell extracts of partially purified urease preparations from such cultures could be elevated by heat treatment, dilution, or dialysis to values comparable to those observed in samples from exponentially grown cells.

Amino Acid Sequence↗

Antigenic variability of bacterial RNA polymerases.

Radioimmunoassay analysis of enteric and some other Gram-negative bacteria has shown that the antigenic structure of the RNA polymerase alpha subunit is more conserved than that of the beta and beta' subunits. Since anti-alpha antibodies do not affect RNA polymerase activity, the constraints which determine the low variability of the antigenic structure of the alpha subunit are not directly related to its functional role. The antigenic determinants of the alpha subunit located on the surface of the RNA polymerase molecule are more conserved than those involved in contacts with other subunits; an opposite tendency characterizes the beta subunit. The range of variability of the antigenic determinants buried inside the RNA polymerase molecule suggests that the subunits are attached to each other rather loosely. Immunological comparison of RNA polymerases provides a simple method for reconstructing bacterial genealogies. The genealogy of the bacteria examined is essentially in agreement with phylogenetic trees based on 16S and 5S rRNA sequence characterization. This argues against extensive interspecific transfer of genes coding for components of the transcription and translation apparatus.

Bacillaceae↗

[Daily bath and its effect on the normal human skin flora quantitative and qualitative investigations of the aerobic skin flora (author's transl)].

The behaviour of the aerobic skin flora of the flexor sides of the forearm, under a daily repeated bath during 21 days, using two marketable bath-supplements, was investigated in 24 volunteers for its quantitative and qualitative aspects. The combined scrubbing-washing-method was used as described by Burtenshaw with Ringer's solution. After the daily baths, using Wilcoxon-test, statistically no significant changes in the total number of microorganizations and in the number of the single groups of bacteria were found in the areas investigated by intermittent samplings of the skin flora. In the analysis of the composition of the normal aerobic skin flora, however, changes were obvious in 4 volunteers. There were no clinical changes of the skin surface in any of the volunteers.

Actinomycetales↗

Ammonium assimilation in Proteus vulgaris, Bacillus pasteurii, and Sporosarcina ureae.

No active uptake of ammonium was detected in Proteus vulgaris, Bacillus pasteurii, and Sporosarcina ureae, which indicates that these bacteria depend on the passive diffusion of ammonia across the cell membrane. In P. vulgaris the glutamine synthetase-glutamate synthase (GS-GOGAT) pathway and glutamate dehydrogenase (GDH) were present, and these enzymes exhibited high affinities for ammonium. In B. pasteurii and S. ureae, however, no GS activity was detected, and GOGAT activity was only present in S. ureae. GDH enzymes were present in these two organisms, but showed only low affinity for ammonium, with apparent Km-values of 55.2 mM in B. pasteurii and 36.7 mM in S. ureae, respectively. These observations explain why P. vulgaris is able to grow at neutral pH and low ammonium concentration (2 mM), while B. pasteurii and S. ureae require high ammonium concentration (40 mM) and alkaline pH for growth.

Alcaligenes↗

Protein secretion in Bacillus brevis.

Many strains of Bacillus brevis were isolated from nature as very efficient producers of extracellular proteins. Strains identified as B. brevis including these protein-hyperproducers were reclassified into at least 6 species according to numerical analysis, DNA base composition, and DNA-DNA hybridization. We developed a host-vector system using appropriate strains of these Bacillus brevis as a host, which is excellent for the secretion of heterologous proteins. Utilizing the powerful promoters and signal peptide-coding regions of the cell wall protein gene, various expression-secretion vectors were constructed. The cell wall protein genes of these B. brevis are transcribed from multiple and tandemly arranged promoters. Transcription from P2, one of the major promoters among them, was enhanced at the early stationary phase of growth, when divalent cations in the medium was depleted and the cell wall protein layers started to be shed. Translation of the cell wall protein gene transcripts starts at the two sites located tandemly in the same reading frame. The two forms of secretory precursors, translation products from the two sites, are cleaved at the same position giving rise to the same mature proteins. The nucleotide sequence from the promoter to the translation start site is highly conserved in protein-hyperproducing B. brevis. For the efficient secretion of some heterologous proteins, protein-hypersecreting mutants had to be selected. The engineering of the signal peptide was also often necessary to obtain a good secretion of heterologous proteins.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗

Anaerobic bacteria: evaluation of disc susceptibility to four cephalosporins.

The disc diffusion technique was evaluated with 178 strains of anaerobes and four cephalosporins (cephalothin, cefamandole, cefazolin and cefoxitin). Good correlation in results was found in comparison with the agar dilution technique (p less than 0.001) with the exception of cefamandole and cefazolin against anaerobic cocci (p greater than 0.05). Choosing a breakpoint of 8 microgram/ml for distinguishing susceptible and resistant strains, we determined corresponding incubation, the rate of error is less than 1% for false susceptible and less than 5% for false resistant. However, some strains of anaerobic cocci required a 48 hour incubation period for allowing visible growth. Moreover, a great deal (60.5%) of overlapping zone diameters made interpretation of disc diffusion test difficult among Bacteroides fragilis strains classed as susceptible, intermediate and resistant occuring with cefoxitin. The results have shown that the cephalothin disk will not accurately predict susceptibility of B. fragilis to cefoxitin.

Bacillaceae↗

Precipitation of minerals by 22 species of moderately halophilic bacteria in artificial marine salts media: influence of salt concentration.

Precipitation of minerals was shown by 22 species of moderately halophilic bacteria in both solid and liquid artificial marine salts media at different concentration and different Mg2+-to-Ca2+ ratio. Precipitation of minerals was observed for all the bacteria used. When salt concentration increased, the quantity and the size of bioliths decreased, the time required for precipitation being increased. The precipitated minerals were calcite, magnesian calcite, aragonite, dolomite, monohydrocalcite, hydromagnesite and struvite in variable proportions, depending on the bacterial species, the salinity and the physical state of the medium; the Mg content of the magnesian calcite also varied according to the same parameters. The precipitated minerals do not correspond exactly to those which could be precipitated inorganically according to the saturation indices. Scanning electron microscopy showed that the formation of the bioliths is initiated by grouping of calcified cells and that the dominant final morphologies were spherulitic with fibrous radiated interiors. It was demonstrated that moderately halophilic bacteria play an active role in the precipitation of carbonates and we hypothesize about this process of biomineralization.

Bacillaceae↗

Inexplicable suppression of hepatic uptake of gallium-67, a case report.

We describe here a case report of a patient with acute lymphocytic leukemia in whom hepatic gallium-67 (Ga-67) uptake was suppressed. The patient was hospitalized with increasing dyspnea. In Ga-67 scintigraphy, there was no hepatic uptake, although other physiological uptake was clearly observed. In addition, the scintigraphy showed increased accumulation in the right lung consistent with infection. We considered possible reasons for these findings. The patient had no history of chemotherapy or blood transfusion, and his iron metabolism was almost normal. He was not receiving any medication which might reduce hepatic blood flow. Blood chemistry suggested normal hepatic and renal function. The patient died from pneumonia 6 weeks later. The autopsy revealed extensive infiltration of the right lung with Bacillus cereus (B. cereus). Metabolic acidosis and/or iron utilization of B. cereus may induce both increased Ga-67 accumulation in the infected lesion and suppressed uptake in the liver, but these mechanisms could not explain normal physiological uptake in the other organs. This case warranted the further study of the hepatic Ga-67 uptake mechanism.

Aged↗