The role of nonsporulating anaerobic bacteria in the etiology of postabortion and postpartal endometritis

As the result of the study carried out by the authors, a conclusion has been made on the leading role of polymicrobial aerobic-anaerobic complexes, capable of producing a synergic effect, in the etiology of endometritis, anaerobic microorganisms prevailing in their importance. In the etiology of endometritis the most important organisms are bacteroids and peptostreptococci among anaerobes, enterobacteria and group D streptococci among aerobes. Anaerobic bacteria causing endometritis are most sensitive to dalacin and lincomycin, least sensitive to benzylpenicillin, resistant to aminoglycosides. The amounts of aerobic and anaerobic microflora contaminating the uterine cavity correlate with the severity of endometritis.     

Effect of Actinomyces pyogenes and gram-negative anaerobic bacteria on the development of bovine pyometra

Fifteen lactating Holstein cows were used in a trial designed to evaluate the effectiveness of intrauterine inoculation (challenge) of Actinomyces pyogenes (A) alone or in combination with Fusobacterium necrophorum (F) and Bacteroides melaninogenicus (B) to induce pyometra. Cows were assigned to one of five groups: A (n = 3), AB (n = 3), AF (n = 3), ABF (n = 3) or C (control, broth medium alone; n = 3). All cows exhibited estrus 12 or 13 d prior to challenge (Day 0=first day of challenge). During the prechallenge period, the reproductive tract of each cow was palpated per rectum and uterine fluid aspirates for culture and uterine biopsies were also obtained. All cows received an intravenous injection of 5,000 IU of human chorionic gonadotropin (hCG; Day 5) and an intrauterine infusion of 40 ml of 0.7% iodine solution (Day 1). Cows were then inoculated on Days 0, 1 and 2 of the experiment. Sequential palpations of the reproductive tracts, samples of uterine fluid for culture and uterine biopsies were performed for a total of 30 d after the first inoculation. A cow was diagnosed as having pyometra when purulent uterine fluid and a corpus luteum were detected by palpation per rectum. The number of cows that developed pyometra in Group A was 2 of 3, in Group AB 3 of 3, in Group AF 3 of 3, in Group ABF 3 of 3 and in Group C 0 of 3. Cows with pyometra did not exhibit estrus. In 7 of 11 cows, pyometra persisted for more than 21 d. In cows that developed pyometra, the same species of bacteria infused into the uterus were usually recovered one or more times during the postchallenge period. When clinical pyometra was diagnosed, histologic endometritis was invariably present. Histologic endometritis and concurrent isolation of A . pyogenes alone or A . pyogenes with gram-negative anaerobic bacteria occurred in 91.7% of samples during the postchallenge period. Regardless of bacterial treatment, gram-negative anaerobic bacteria were frequently isolated with A . pyogenes during this period.     

Rapid methods for differentiating gram-positive from gram-negative aerobic and facultative anaerobic bacteria

Different tests based on lysis by KOH and on reaction with fluorogenic and chromogenic substrates, L-alanine-4-nitroanilide (LANA); L-alanine-4-methoxy- beta-naphthylamide (MNA); 4-alanine-2-amidoacridone (AAA); L-alanine-7-amido- 4-methylcoumarin (AAMC); 8-anilino-1-naphthalene-sulphonic acid (ANS) were compared for their suitability to distinguish Gram-positive from Gram-negative bacteria. A concentration of 100 micrograms/ml was chosen for incorporating LANA, AAA, AAMC and ANS into the growth medium, based on sensitivity tests. MNA did not show any detectable reaction over a concentration range from 50 to 200 micrograms/ml, and led to inhibition of all bacteria at 200 micrograms/ml. In the examination of a total of 146 bacterial strains, including Yersinia enterocolitica, Bacillus cereus, and B. subtilis the KOH test was not comparable with the Gram staining. A good correlation with Gram staining was found between LANA, AAA and AAMC added to plate count agar on one hand, and LANA and AAMC impregnated paper strips on the other hand, thereby utilizing the aminopeptidase activity. Agar containing ANS showed detectable fluorescence with all Gram-negative strains, but with Staphylococcus aureus and Staph. epidermidis a weak reaction was also observed. AAMC was selected for a rapid paper strip test. With this substrate a pronounced blue fluorescence was obtained with Gram-negative colonies.     

高效厌氧纤维素降解细菌的分离及酶特性研究

采用透明圈初筛和滤纸降解率复筛的方法从内蒙古绵羊瘤胃内容物中分离到高效厌氧纤维素降解细菌4株.通过形态学、生理生化反应、生态特性和遗传型的鉴定,所分离的4株菌WHQ、LYQ、LBG-1和NDF-3分别归为溶纤维丁酸弧菌(Butyrivibrio fibrisollvens)、黄色瘤胃球菌(Ruminococcus flavefaciens)、产琥珀酸丝状杆菌(Fibrobacter succinogenes)和解多糖梭菌(Clostridium polysaccharolyticum).测定了4株菌对滤纸的降解率,WHQ、LYQ、LBG-1和NDF-3的2周滤纸降解率分别为25.1%、14.3%、21.0%和20.6%.本研究同时对4株菌的滤纸酶活力、羧甲基纤维素酶活力和β-葡萄糖苷酶活力进行了测定.     

Bile acid biotransformation rates of selected gram-positive and gram-negative intestinal anaerobic bacteria.

Treatment of whole cell suspensions of $\text{Eubacterium aerofaciens}$ and $\text{Bacteroides fragilis}$ with lysozyme resulted in a marked increase (>100-fold) in the rates of biotransformation of cholate to 7-ketodeoxycholate (7-KD) in the former but only a 2-fold increase in the latter bacterium. In $\text{B. fragilis}$ the total activity of both NAD-dependent 7-α-hydroxysteroid dehydrogenase (7-α-OHSDH) and bile salt hydrolase (BSH) increase markedly during the stationary growth phase. Both enzymes were found in the spheroplast lysate and the Triton-soluble washed membrane fractions but only BSH was found in the spheroplast medium.     

Determination of diamino acids in peptidoglycans from anaerobic bacteria

Summary. Peptidoglycans isolated from two Fusobacterium species of anaerobic bacteria were analyzed for constituent amino acids. Hydrolysis conditions were varied to optimize the yield of diamino acids from peptidoglycan. The o-phthalaldehyde derivatives of the diamino acid stereoisomers were separated by high-performance liquid chromatography (OPA-HPLC), and variations in the relative areas of the two peaks noticed during analysis of solid samples were attributed to sampling errors. Co-derivatization/injection experiments using standards of the meso and rac forms separated from commercial mixtures demonstrated that meso-2,6-diaminopimelic acid and meso-lanthionine were peptidoglycan components in Fusobacterium varium and Fusobacterium nucleatum, respectively. The protonated molecules of 2,6-diaminopimelic acid and lanthionine were detected in peptidoglycan hydrolyzates by off-line, flow-injection electrospray mass spectrometry (ESI-MS). In ESI-MS-MS experiments under identical collision-induced dissociation (CID) conditions, peptidoglycan-derived and standard diamino acids exhibited similar fragmentations. Fragmentation pathways are proposed for each diamino acid. The results confirm that the meso forms of two different diamino acids are utilized in the peptidoglycans of Fusobacterium species. Received February 16, 2001 Accepted May 10, 2001     

Membrane lipids of mesophilic anaerobic bacteria thriving in peats have typical archaeal traits

The 16S ribosomal DNA based distinction between the bacterial and archaeal domains of life is strongly supported by the membrane lipid composition of the two domains; Bacteria generally contain dialkyl glycerol diester lipids, whereas Archaea produce isoprenoid dialkyl glycerol diether and membrane-spanning glycerol dialkyl glycerol tetraether (GDGT) lipids. Here we show that a new group of ecologically abundant membrane-spanning GDGT lipids, containing branched instead of isoprenoid carbon skeletons, are of a bacterial origin. This was revealed by examining the stereochemistry of the glycerol moieties of those branched tetraether membrane lipids, which was found to be the bacterial 1,2-di-O-alkyl-sn-glycerol stereoconfiguration and not the 2,3-di-O-alkyl-sn-glycerol stereoconfiguration as in archaeal membrane lipids. In addition, unequivocal evidence for the presence of cyclopentyl moieties in these bacterial membrane lipids was obtained by NMR. The biochemical traits of biosynthesis of tetraether membrane lipids and the formation of cyclopentyl moieties through internal cyclization, which were thought to be specific for the archaeal lineage of descent, thus also occur in the bacterial domain of life.     

Determination of mercury and organomercurial resistance in obligate anaerobic bacteria

A methodology for determining the minimum inhibitory concentration of inorganic and organomercurial compounds for obligate anaerobic bacteria is described. A wide variation in the susceptibility of anaerobic clinical and sewage isolates was observed. Isolates of Bacteroides ruminicola and Clostridium perfringens resistant to mercury were examined for their plasmid content and ability to demonstrate inducible resistance. None of the resistant anaerobes contained any plasmids, while resistant facultative isolates from the same source contained several plasmids. In 24 h, resistant strains of clostridia and Bacteroides volatilized 20 and 43% of the 203Hg2+ added to cultures, while Escherichia coli R100 and a sewage isolate of Enterobacter cloacae volatilized 63 and 27%, respectively, of the added 203Hg2+. Attempts to induce mercury resistance in the aerobic isolates were successful, but no induction was seen in the anaerobes. Thus, mercury resistance in these anaerobic isolates was neither inducible nor plasmid mediated.     

Anhydrobiotic engineering of gram-negative bacteria

Anhydrobiotic engineering aims to improve desiccation tolerance in living organisms by adopting the strategies of anhydrobiosis. This was achieved for Escherichia coli and Pseudomonas putida by osmotic induction of intracellular trehalose synthesis and by drying from trehalose solutions, resulting in long-term viability in the dried state.     

Lyophilization of nonsporulating strains of the fungusClaviceps

Summary Maintenance of active strains of microorganisms is one of the fundamental problems for every successful industrial biosynthesis. In this paper, the efficiency of lyophilization for long-term preservation of nonsporulating strains of the fungusClaviceps, producing ergot alkaloids in saprophytic cultures, was studied. It was found that lyophilization is a suitable method for the preservation of the testedClaviceps strains. All treated strains have retained their vitality and biochemical properties unchanged after three years of storage.     

Antibiotic sensitivity of nonfermenting gram-negative bacteria

Antibiotic resistance of 132 strains of nonfermenting gram-negative bacteria (NGNB) was studied. 43, 20, 17, 14 and 12 of them belonged to Acinetobacter calcoaceticus (anitratus and lwoffi), Pseudomonas cepacia, Alcaligenes faecalis, P. stutzeri and P. maltophilia, respectively. With rare exceptions all the strains were resistant to benzylpenicillin, oxacillin, lincomycin, ampicillin and cephaloridine. Sensitivity to the other antibiotics varied within wide ranges. Amikacin (94.3 per cent) and tobramycin (90.8 per cent), as well as polymyxin, rifampicin and gentamicin (71.7-66.9 per cent) had the highest effect. The majority of the antibiotics had higher activity (p less than 0.01) against the tested NGNB as compared to their activity against P. aeruginosa. Antibioticograms of every of the tested species of NGNB revealed that P. cepacia and P. stutzeri were the most resistant species. The biovars of Acinetobacter varied in their antibiotic resistance: A. subsp. lwoffi was more sensitive to the majority of the antibiotics though some of them, i.e. doxycycline, carbenicillin, and polymyxin were more active against A. subsp. anitratus.     

Polymorphic behavior of gram-negative bacteria membranes

Summary Freeze-fracture and ultrathin section electron microscopy as well as³¹P-NMR spectroscopy and light scattering ofEscherichia coli andPseudomonas putida cells under conditions promoting the ability of cells to take up exogenous DNA's (high concentrations of divalent cations and a specific temperature regime) reveal the extensive polymorphic changes and the formation of various structural defects in cellular membranes. Polymorphic changes occur during the heat shock at 42 to 44°C of the cells preincubated at 0°C in the presence of high concentration of Ca²⁺ or Ba²⁺ cations and include the formation of various vesicle- and tube-like structures, intermembrane and intercellular contacts followed by membrane fusion and sometimes even by cell fusion. The results obtained suggest the occurrence of phospholipid-enriched zones in the outer leaflet ofE. coli outer membrane. This suggestion is verified and confirmed with the help of phospholipase C, a specific phospholipid binding and digesting enzyme. The presented experimental evidence directly supports the suggestion of Ahkong et al. (Nature253:194–195, 1975) on the identity of the mechanisms of membrane contact formation and membrane fusion in model and cellular membranes. The biological relevance of the polymorphic changes observed is shortly discussed.     

Acquisition of siderophores in gram-negative bacteria

The outer membrane of Gram-negative bacteria constitutes a permeability barrier that protects the cell from exterior hazards, but also complicates the uptake of nutrients. In the case of iron, the challenge is even greater, because of the scarcity of this indispensable element in the cell's surroundings. To solve this dilemma, bacteria have evolved sophisticated mechanisms whereby the concerted actions of receptor, transporter and energy-transducing proteins ensure that there is a sufficient supply of iron-containing compounds, such as siderophores.     

Studies on anaerobic bacteria

Summary A new chromogenic anaerobe, Clostridium roseum nov. spec., has been found. It is characterized by: red-orange pigment, turning purplish on oxidation; gelatin liquefaction and other evidence of proteolysis; nitrate reduction; fermentation of various carbohydrates including pectin; close resemblance to Cl. acetobutylicum in corn mash fermentation, with the same neutral products, acetone, ethyl alcohol and butyl alcohol, in nearly the same ratios; agglutinative specificity and separation from Cl. acetobutylicum and Cl. felsineum, as well as several less nearly physiologically related butyric anaerobes.