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Identification of some uncommon monounsaturated fatty acids of bacteria.

Location of the double-bond position of monounsaturated fatty acids of various bacteria was accomplished with combined gas chromatography-mass spectrometry analysis of dimethyl disulfide (DMDS) derivatives. The monoenoic fatty acids from whole cells were converted to methyl esters and then to DMDS adducts and analyzed by capillary gas chromatography-mass spectrometry. The mass spectra of DMDS adducts gave an easily recognizable molecular ion and two major diagnostic ions attributable to fragmentation between the two CH3S groups located at the original site of unsaturation. Twenty-one relatively novel monoenoic fatty acids were identified among the bacteria studied. All Flavobacterium species contained i17:1 omega 8c, Bacillus alvei contained i16:1 omega 11c and i17:1 omega 12c, and Psychrobacter immobilis contained 12:1 omega 9c. Resolution of cis and trans isomers with capillary gas chromatography and subsequent mass spectrometry permitted positive identification of 16:1 omega 7c and 16:1 omega 7t in Arcobacter (Campylobacter) cryaerophila and 16:1 omega 9c and 16:1 omega 9t in Aerococcus viridans.

Bacteria↗

DNA relatedness, phenotypic characteristics, and antimicrobial susceptibilities of Globicatella sanguinis strains.

DNA-DNA reassociation was performed on 15 strains of Globicatella sanguinis to compare their taxonomic status with phenotypic characterization. All 15 strains selected for DNA-DNA reassociation readily met the criteria for species relatedness. The relative binding ratio was 81% or greater at the optimal temperature and 76% or greater at the stringent temperature, and the divergence was less than 3% for all strains hybridized with the type strain. These strains included nine strains from the Centers for Disease Control Streptococcus Laboratory culture collection that were previously included in comparative 16S rRNA gene sequencing studies as well as six additional phenotypically variant isolates. DNA-DNA relatedness was less than 18% at the optimal reassociation temperature to Aerococcus viridans, Enterococcus avium, and Streptococcus uberis, which are phenotypically similar to G. sanguinis. This study confirms these Globicatella strains were previously misidentified as S. uberis or S. uberis-like strains based on biochemical characteristics. The biochemical data from 28 strains was compiled to further define the phenotypic criteria for identification of this species. A revised description of the species should be variable reaction for pyrrolidonylarylamidase production (75% positive), positive reaction for the bile esculin test (100%), growth at 45 degrees C (96%), variable reaction for acid production from arabinose (45% positive), and negative starch hydrolysis (0% positive). We also evaluated four rapid identification systems, the Biomerieux rapid ID32 STREP (ID32), the Crystal rapid gram-positive identification (Cry4), the BBL Crystal gram-positive identification (Cry24), and the Remel IDS RapID STR (IDS) systems for their ability to identify these strains.

Adult↗

In vitro antimicrobial activity of gatifloxacin compared with other quinolones against clinical isolates from cancer patients.

Owing to the predominance of gram-positive pathogens in neutropenic cancer patients, newer generation quinolones with an expanded gram-positive spectrum and enhanced potency, may have a role to play for prophylaxis and/or empiric therapy in such patients. The in vitro activity of gatifloxacin was compared with that of ciprofloxacin, levofloxacin and trovafloxacin against 848 recent clinical isolates from cancer patients. Against gram-positive organisms, gatifloxacin was the most active agent tested inhibiting all Aerococcus, Listeria monocytogens, Micrococcus, Stomatococcus mucilaginous, Bacillus, and Rhodococcus equi strains at < or =2 mg/l, its designated susceptibility breakpoint. It was also very active against methicillin-susceptible staphylococci and Streptococcus spp. (including penicillin nonsusceptible Streptococcus pneumoniae and viridans streptococci). It had moderate activity against methicillin-resistant staphylococci and Enterococcus faecalis, inhibiting 68-80% of these strains at < or =2 mg/l. Gatifloxacin also had good activity against the Enterobacteriaceae (although ciprofloxacin was more potent) inhibiting >95% of isolates at < or =1 mg/l. Nonfermentative gram-negative organisms were less susceptible to all 4 agents. Gatifloxacin was very active against Acinetobacter lwoffi (MIC100 0.12 mg/l) and had moderate activity against Acinetobacter baumanii, Chryseobacterium spp., Stenotrophomonas maltophilia, Pseudomonas aeruginosa and other Pseudomonas species. Alcaligenes xylosoxidans strains were relatively resistant to all 4 agents.

Anti-Bacterial Agents↗

Evaluation of five selective media for isolation of catalase-negative gram-positive cocci from bulk tank milk.

Five selective media including Edwards modified medium, Edwards modified medium supplemented with colistin sulfate (5 mg/L) and oxolinic acid (2.5 mg/L), Streptococcus selective medium, Streptosel agar, and thallium-crystal violet-toxin-ferric citrate medium were evaluated for the isolation of streptococci and streptococci-like organisms from raw milk. The sensitivity and specificity of these selective media for streptococci and streptococci-like organisms were determined by using American Type Culture Collection reference strains. Under experimental conditions Edwards modified medium with colistin sulfate (5 mg/L) and oxolinic acid (2.5 mg/L) showed the highest sensitivity (100%) and specificity (100%) for streptococci and streptococci-like organisms followed by thallium-crystal violettoxin-ferric citrate medium, Edwards modified medium, Streptococcus selective medium, and Streptosel agar. Edwards modified medium supplemented with colistin sulfate (5 mg/L) and oxolinic acid (2.5 mg/L) allowed growth of all streptococci and streptococci-like organisms, while inhibiting growth of the staphylococci and gram-negative reference strains. Bulk tank milk samples from 114 dairy herds were spiral plated onto Edwards modified medium with colistin sulfate (5 mg/L) and oxolinic acid (2.5 mg/L). A total of 344 isolates (at least three isolates from each sample) were randomly selected and identified to their species. This medium permitted growth of 328 streptococci and streptococci-like organisms belonging to genera Aerococcus, Enterococcus, Gemella, Lactococcus, Streptococcus, and Vagococcus. When Edwards modified medium supplemented with colistin sulfate (5 mg/L) and oxolinic acid (2.5 mg/L) was evaluated using bulk tank milk samples, the sensitivity and specificity of this medium for streptococci and streptococci-like organisms were observed to be 100 and 87.5%, respectively. The positive predictive value for streptococci and streptococci-like organisms was observed to be 99.4%. The results of the study indicate that Edwards modified medium supplemented with colistin sulfate (5 mg/L) and oxolinic acid (2.5 mg/L) can be used as a selective medium for the isolation of streptococci and streptococci-like organisms from bulk tank milk.

Animals↗

Minimum inhibitory concentrations and disk diffusion zone diameter for selected antibiotics against streptococci isolated from bovine intramammary infections.

The antimicrobial susceptibility to 11 antibiotics was determined of 358 streptococcal isolates representing six species was determined. All isolates were from milk samples obtained from clinical or subclinical cases of bovine mastitis. Agar disk diffusion zone diameters and minimum inhibitory concentrations were obtained for all isolates. Of the antibiotics tested that are available for mastitis therapy, cephalothin, penicillin, ampicillin, novobiocin, and erythromycin demonstrated acceptable activity against nonenterococcal streptococci, but only penicillin and ampicillin demonstrated moderate activity against Enterococcus faecalis. Enterococcus faecalis demonstrated substantial resistance to cephalothin, erythromycin, gentamicin, streptomycin, and tetracycline, while the nonenterococcal streptococci demonstrated substantial resistance to gentamicin, streptomycin, and tetracycline. Streptococcus uberis demonstrated greater antibiotic resistance than did Streptococcus dysgalactiae, while Aerococcus viridans demonstrated the greatest overall resistance of the nonenterococcal species.

Animals↗

Immunogen-dependent quantitative and qualitative differences in phagocytic responses of the circulating hemocytes of the lobster Homarus americanus.

Phagocytic responses in circulating hemocytes of the lobster Homarus americanus were measured before and after treatment of lobsters with 2 different immunogens: (1) lipolysaccharide (LPS) or endotoxin from a non-pathogenic Pseudomonas perolens, and (2) a vancomycin/live Gram-positive pathogen (Aerococcus viridans [var.] homari) combination, essentially attenuated cells, shown previously to induce a high degree of resistance to this pathogen. The responses elicited by each of the immunogens were markedly different. Hemocytes drawn from LPS-treated lobsters showed significant, largely non-specific, increases in phagocytic responses over baseline values against sheep red blood cells and an array of test bacteria, with the notable exception of the pathogen. In marked contrast, induction with the vancomycin/live pathogen combination resulted in highly significant and specific increases in phagocytic responses to the pathogen and to the related, (but avirulent) strains of the pathogen, as well as inducing in the lobsters the usual high degree of resistance to the pathogen. These results suggest that quantitative and qualitative variations in phagocytic and resistance levels induced in at least 1 crustacean genus are determined largely by the particular characteristics of the immunogen.

Animals↗

Microbial colonization of tourniquets used in orthopedic surgery.

This study analyzed tourniquets used for orthopedic surgery in our hospital to determine the frequency and type of microbial contamination. Group A tourniquets were from our main operating room, Group B tourniquets were from our ambulatory surgicenter, Group C tourniquets were unused, prepackaged, sterile tourniquets from our main operating room, and Group D tourniquets were sterilely packed tourniquets from our ambulatory surgicenter. Tourniquets from Groups A, B, C, and D had 100%, 40%, 0%, and 0% microbial growth, respectively. For Group A tourniquets, coagulase-negative staphylococci, Bacillus, and Staphylococcus aureus were present in 100%, 60%, and 20% of tourniquets, respectively. Twenty percent were contaminated either with Streptococcus sanguis, Aerococcus viridans, or Cornyebacterium species. Coagulase-negative staphylococci and Bacillus were present in 40% and 30% of Group B tourniquets, respectively. Tourniquet contamination may be a risk factor for the development of surgical site infection in orthopedic surgery.

Equipment Contamination↗

Activation of the lectin complement pathway by H-ficolin (Hakata antigen).

Ficolins are a group of proteins which consist of a collagen-like domain and a fibrinogen-like domain. In human serum, there are two types of ficolins named L-ficolin/P35 and H-ficolin (Hakata Ag), both of which have lectin activity. We recently demonstrated that L-ficolin/P35 is associated with mannose-binding lectin (MBL)-associated serine proteases (MASP) 1 and 2 and small MBL-associated protein (sMAP), and that the complex activates the lectin pathway. In this study, we report the characterization of H-ficolin in terms of its ability to activate complement. Western blotting analysis showed the presence of MASP-1, MASP-2, MASP-3, and sMAP in H-ficolin preparations isolated from Cohn Fraction III. The MASPs in the preparations had proteolytic activities against C4, C2, and C3 in the fluid phase. When H-ficolin preparations were bound to anti-H-ficolin Ab which had been coated on ELISA plates, they activated C4, although no C4 activation was noted when anti-MBL and anti-L-ficolin/P35 were used. H-ficolin binds to PSA, a polysaccharide produced by Aerococcus viridans. C4 was activated by H-ficolin preparations bound to PSA which had been coated on ELISA plates. These results indicate that H-ficolin is a second ficolin which is associated with MASPs and sMAP, and which activates the lectin pathway.

Antibodies, Monoclonal↗

Mastitis-causing streptococci are important contributors to bacterial counts in raw bulk tank milk.

The objective of this study was to probe the contribution of streptococci to the microbial quality of raw milk. Over a 5-month period, bulk tank milk samples from 48 New York State dairy farms were analyzed qualitatively for bacterial ecology and quantitatively for total bacterial, streptococcal, staphylococcal, and gram-negative bacterial counts. Linear regression analysis was used to determine the contribution of differential counts to total bacterial counts. Streptococci, staphylococci, and gram-negative bacteria accounted for 69, 3, and 3% of total bacterial count variability, respectively. Randomly selected Streptococcus isolates from each bulk tank milk sample were identified to species by means of the API 20 STREP identification system. The most commonly identified streptococcal species were Streptococcus uberis, Aerococcus viridans, and Streptococcus agalactiae, which were detected in 81, 50, and 31% of 48 bulk tank samples, respectively. For five herds, S. uberis isolates from bulk tank milk and individual cows were characterized by PvuII ribotyping. A farm-specific dominant ribotype was found in each bulk tank sample, and that ribotype was isolated from at least one cow within each herd of origin. Bacteriological and strain typing data indicate that control of streptococci, specifically mastitis-causing species, is important for improvement of the microbial quality of raw milk in New York State.

Animals↗

PCR detection of Bacillus and Staphylococcus in various foods.

A broad-range PCR assay for the detection of bacteria belonging to Bacillus and Staphylococcus genera was developed. Primers targeting the bacterial 16S rRNA gene were newly designed and used in a PCR assay. To determine the specificity of the assay, 81 different bacterial strains (of 50 genera), 2 fungi, 3 animals, and 4 plants were tested. Results were positive for every tested Bacillus, Staphylococcus, or Aerococcus strain. In addition, the result for Listeria grayi was positive with lower PCR product. For all other bacterial strains and eukaryotes tested, results were negative. Bacterial DNA was prepared with the use of achromopeptidase and Chelex 100 resin from culture after growth in brain heart infusion medium. To test the sensitivity of this PCR assay for Bacillus or Staphylococcus genus, either Bacillus cereus or Staphylococcus aureus was inoculated into various foods with undetectable levels of endogenous microbial contamination as an indicator. Inoculation of bacteria at 10 to 30 CFU/g of food was followed by a 5-h enrichment culture step after which the PCR assay allowed the detection of bacterial cells. When the inoculation (B. cereus or S. aureus) of 10 to 90 CFU/g into noodle foods containing endogenous microflora (10(3) to 10(5) CFU/g) was followed by a 6-h enrichment culture step, the PCR assay detected the bacteria. Including the enrichment culture step, the entire PCR detection process can be completed within 8.5 h.

Bacillus↗

Bacterial flora of the conjunctiva and nasal cavity in normal and diseased captive bustards.

A survey was carried out to describe the normal aerobic bacterial flora of the conjunctiva and nasal cavity of captive houbara bustards (Chlamydotis undulata), kori bustards (Ardeotis kori), and white-bellied bustards (Eupodotis senegalensis) maintained at the National Avian Research Center, Abu Dhabi, United Arab Emirates. A total of 58 samples were examined from the nasal cavity and 55 samples from the conjunctiva of healthy bustards. There was no bacterial growth in 45% of conjunctival samples. Bacteria isolated from the conjunctiva of healthy birds included Micrococcus spp., Staphylococcus auricularis, Staphylococcus xylosus, Staphylococcus capitis, Staphylococcus warneri, Bacillus spp., and Enterobacter amigenus. Bacteria isolated from the nasal cavity of healthy birds included Bacillus spp., Micrococcus spp., S. auricularis, S. xylosus, Staphylococcus simulans, Staphylococcus saprophyticus, Staphylococcus hyicus, Staphylococcus cohnii, Staphylococcus sciuri, Aerococcus spp., and Providencia rettgeri. These findings were compared with bacterial isolates from bustards with clinical signs of ocular or upper respiratory tract diseases. Mycoplasma spp., Pseudomonas aeruginosa, Pseudomonas stutzeri, Proteus mirabilis, Escherichia coli, Klebsiella spp., Aeromonas hydrophila, and Staphylococcus aureus were the pathogenic bacteria isolated from the conjunctiva of 34.3% bustards with ocular discharges. Mycoplasma spp., P. aeruginosa, Pseudomonas spp., P. mirabilis, E. coli, Klebsiella pneumoniae, and S. aureus were the pathogenic bacteria isolated from the nasal cavity of 74% bustards with upper respiratory tract diseases.

Animals↗

[Hemopericardium caused by endocarditis ulcerosa: an unusual cause of sudden death in adolescence].

The authors report on the case of a 18-year-old student who died suddenly and unexpectedly of a pericardial tamponade after perforated ulcer-polypous endocarditis of the aortic valve. The detection of aerococcus urinae--which usually causes urinary tract infection--on the inflamed aortic valve demonstrates an unusual pathogenetic chain linking complexes of findings in the urogenital system and the heart.

Adolescent↗

The response of the lymphatic system to the human skin resident bacteria.

We designed a study to identify skin bacterial strains producing most intensive changes in lymphoid tissue of the draining nodes and analyse the node cell phenotypes reacting to bacterial antigens. The obtained results were expected to help in establishing rational therapy in recipients of hand allografts. The highest level of activation of lymphoid tissue was found after inoculation with S. epidermidis, Acinetobacter, Enterobacter and Aerococcus. There was an increase in weight and cell concentration of nodes, intense expression of W3/25+ (CD4), OX6+ (class II), EDI+ (CD14), CD54+ and OX62+ cells in the paracortex and medulla 7 days after bacterial inoculation. The most advanced changes were seen after recall bacterial stimulation. Taken together, evident differences in the intensity of response to the investigated bacterial strains were found. The obtained information can be useful for establishing rational antibiotic therapy after composite graft transplantation.

Animals↗

[Use of the Streptotest for identification of enterococci and viridans-type streptococci].

The new identification system STREPTOtest (fy. Lachema, Brno) was evaluated. A total 118 well-known strains of genus Enterococcus, Streptococcus, Aerococcus, Stomatococcus and Gemella were tested. The STREPTOtest system rapidly and reliably distinguished genus Enterococcus from genus Streptococcus. This method identified correctly to the species level 65.9% of enterococci strains and 52.5% of streptococci strains. The STREPTOtest was used for identification of A. viridans and S. mucilaginosus and it was possible to separate these strains from similar ones. A new differentiation chart for identification of all 13 recently described enterococcal species was proposed.

Bacteriological Techniques↗

[Gram-positive cocci as an opportunistic infection factor].

The frequency of opportunistic infections increased in the last years. The opportunistic infections are serious and very difficult to diagnose complication at patients with lower immunity. The following factors are conducive to develop infections caused by opportunistic bacteria: invasive research methods, medical treatments and immunological defects. The publication presents pathogenicity of bacteria of following types: Aerococcus, Kocuria, Kytococcus, Lactococcus, Leuconostoc, Micrococcus, Pediococcus. There are also mentioned diagnostic and treatment issues in the publication. The purpose of this study is to draw attention to infection caused by opportunistic bacteria, which are ingredients of physiological flora as well as diagnostic difficulties, which occur in case of isolation of this pathogens. Quick and correct diagnosis of etiological factor and applying of proper treatment can have very important impact on effectiveness of therapy and decrease of mortality in this kind of infections.

Gram-Positive Cocci↗

Sensitivity of oral bacteria to imipenem and other antibiotics.

Specimens from a variety of oral and maxillofacial infections were cultured and 88 bacterial isolates belonging to the genera Bacteroides, Actinomyces, Staphylococcus, Veillonella, Eubacterium, Streptococcus and Aerococcus were obtained. These were tested for their sensitivity to imipenem, penicillin, cefuroxime, vancomycin, erythromycin and fusidic acid. Imipenem proved to be the most effective of the antibiotics tested, inhibiting all 88 isolates. Penicillin was the least effective in that it inhibited only 56% of the isolates.

Anti-Bacterial Agents↗

[Septic arthritis as an initial manifestation of endocarditis].

A 60-year-old female patient was admitted to the hospital with high fever and an acute onset of pauciarticular arthritis. Clinical examination and ultrasound imaging of the right knee demonstrated a severe synovitis with a large synovial effusion and a Baker's cyst. Arthrocentesis showed an opaque viscous synovial fluid with a highly elevated white blood cell count. In synovial fluid cultures and in serial blood cultures Aerococcus viridans (Av) was detected. Further examination revealed a mitral valve endocarditis as the origin of the septicemia. This case report is the first one to describe the association of an endocarditis and a septic arthritis due to Av.

Arthritis, Infectious↗

Teicoplanin for therapy of gram-positive infections in neutropenic patients.

Teicoplanin was evaluated in 20 febrile neutropenic patients as additional treatment for suspected Gram-positive infections after inadequate response to initial empiric ceftazidime monotherapy. Five patients with primary septicaemia, four with secondary septicaemia, 12 with localized infections and three with pyrexia of unknown origin were treated with teicoplanin (200 mg bolus intravenously, once daily after a 400 mg loading dose), whilst ceftazidime (2 g, 8-hourly, 30 min infusions) was continued. Four patients were unassessable (tuberculous, viral, protocol violation, and non-infectious pyrexial episode). Clinical cure for the combination was achieved in 11 or the 16 assessable cases (69%). Ten of the eleven (91%) bacteriologically confirmed infections were cured after addition of teicoplanin. Three strains of Staphylococcus aureus, four of Staphylococcus epidermidis (two methicillin resistant), and three strains of Streptococcus faecalis were isolated from successfully treated patients. One patient with Aerococcus and Enterobacter cloacae infection only improved after addition of erythromycin. One superinfection occurred with signs of interstitial pneumonitis in a patient following bone marrow transplantation. Neither ototoxicity nor nephrotoxicity occurred during treatment. A transient rise of liver transaminases was observed in four patients, but was attributable to teicoplanin in only one case. It is concluded that teicoplanin is a potentially effective and well-tolerated antimicrobial agent in neutropenic patients with infections due to Gram-positive organisms.

Agranulocytosis↗