[Accurate diagnosis of Pseudomonas luteola in routine microbiology laboratory: on the occasion of two isolates]. / Rutin mikrobiyoloji laboratuvarinda Pseudomonas luteola'nin dogru tanisi: Iki sus nedeniyle.

Pseudomonas luteola which was previously known as Chryseomonas luteola; is a gram-negative, non-fermentative, aerobic, motile, non-spore-forming bacillus. It is frequently found as a saprophyte in soil, water and other damp environments and is an opportunistic pathogen in patients with underlying medical disorders or with indwelling catheters. It has been reported as an uncommon cause of bacteremia, sepsis, septic arthritis, meningitis, endocarditis, and peritonitis. Thus, early and accurate identification of this rare species is important for the treatment and also to provide information about the epidemiology of P.luteola infections. This report was aimed to draw attention to the accurate identification of P.luteola in clinical samples, upon the isolation and identification in two cases in the medical microbiology laboratory of a university hospital. In February 2011, a 66-year-old man, with chronic obstructive pulmonary disease, coronary artery disease and aplastic anemia, was admitted to our hospital due to progressive dyspnea. A chest tube was inserted on the 20th day of admission by the reason of recurrent pleural effusion. Staphylococcus aureus and a non-fermentative gram-negative bacillus (NFGNB) with wrinkled, sticky yellow colonies were isolated from the pleural fluid sample obtained on the 9th day following the insertion of the chest tube. In February 2012, a 7-year-old male cystic fibrosis patient who had no signs and symptoms of acute pulmonary exacerbation was admitted to the hospital for a routine control. This patient had chronic colonization with Pseudomonas aeruginosa and S.aureus and his sputum sample obtained at this visit revealed isolation of P.aeruginosa, S.aureus, Aspergillus fumigatus and a wrinkled, sticky yellow NFGNB. Both of these NFGNB were identified as P.luteola by the Phoenix automated microbial identification system (BD Diagnostics, USA). To evaluate the microbiological characteristics of these two isolates, the strains were further analysed by VITEK MS (bioMerieux, France) and Microflex LT mass spectrometer (Bruker Daltonics, Germany). Both of the MALDI-TOF-MS systems identified the isolates as P.luteola and 16S rRNA gene sequencing (ABI PRISM 3100, Applied Biosystems, USA) also confirmed the identification. The strains had wrinkled, sticky yellow colonies which were oxidase-negative, catalase-positive and non-fermentative. The Gram stained smears of the colonies revealed clusters of gram-negative bacilli probably embedded into a biofilm matrix. Since there are no accepted standards for testing the antibiotic susceptibility of P.luteola strains, the standards determined by CLSI for "other non-Enterobacteriaceae" (non-fermentative bacteria excluding P.aeruginosa, Acinetobacter spp., Burkholderia cepacia, B.mallei, B.pseudomallei and Stenotrophomonas maltophilia) were used for the susceptibility testing. Gradient MIC method (E-Test, bioMerieux, France) revealed that the isolates were susceptible to gentamicin, piperacillin-tazobactam, ceftazidime, cefepime, meropenem, colistin and levofloxacin. Accurate and prompt identification of P.luteola which is identified as a rare pathogen in serious cases is of critical importance since it has been suggested that this organism is likely to become more frequent as a nosocomial pathogen since the interventional processes increase in current medical practice. This report supported that Phoenix automated phenotypic identification system (BD Diagnostics, USA) and the two MALDI-TOF-MS based systems (VITEK MS and Bruker Microflex LT mass spectrometer) were successfull in the accurate identification of P.luteola.

Effect of Lonicera japonica extract on Mycoplasma gallisepticum in naturally infected broiler flocks.

1. In this study, the effect of chlorogenic acid extract from Lonicera japonica Thunb. on Mycoplasma gallisepticum infections and the performance of broiler flocks was investigated. 2. A total of 360 Ross-308 broiler chicks taken from M. gallisepticum seropositive flocks were divided equally into three groups designated as control (nothing administered), antibiotic (Tylosin tartrate given for the first 3 d and d 20-22) and test group (chlorogenic acid extract given twice a day on d 16 and 22). 3. Broiler performance analysis, serological tests (slide agglutination), molecular identification (polymerase chain reaction) and histopathological examination were performed to detect M. gallisepticum. 4. The results show that chlorogenic acid not only increases live body weight but is also an alternative treatment option in M. gallisepticum-infected broiler flocks.

Emerging pathogen Arcobacter spp. in acute gastroenteritis: molecular identification, antibiotic susceptibilities and genotyping of the isolated arcobacters.

The aims of this study were as follows: (i) to isolate Arcobacter spp. from the stool samples of patients with gastroenteritis; (ii) to identify them with molecular methods; (iii) to genotype them using enterobacterial repetitive intergenic consensus (ERIC)-PCR; and (iv) to determine their antibiotic susceptibilities. For the study, a total of 3287 diarrhoeal stool samples submitted to the Microbiology Laboratory of the Kayseri Training and Research Hospital, Kayseri, Turkey, between 2010 and 2011 were analysed. Campylobacter blood-free selective medium supplemented with cefoperazone, amphotericin B and teicoplanin was used for isolation. Medium inoculated with stool samples was incubated microaerobically at 37 °C for 72-96 h. Phenotypic tests, a genus-specific PCR and a multiplex PCR were used to identify the arcobacters, whilst ERIC-PCR was used for genotyping and the antibiotic susceptibilities of the isolates were detected by E-test. Arcobacter spp. were isolated from nine of the 3287 samples. These nine isolates were identified as Arcobacter butzleri and all showed different ERIC-PCR profiles. All nine isolates were resistant to ampicillin and susceptible to gentamicin, tetracycline, erythromycin and ciprofloxacin. As far as is known, this is the first study in which A. butzleri has been isolated from human acute gastrointestinal infections in Turkey. According to these results, it is recommended that, when investigating the aetiology of infections of the digestive system in humans, Arcobacter spp. be considered for inclusion. The results of this study should contribute to our knowledge related to A. butzleri infections in humans.

N-Alkyl-N-arylmethylpiperidin-4-amines: novel dual inhibitors of serotonin and norepinephrine reuptake.

A series of N-alkyl-N-arylmethylpiperidin-4-amines have been prepared and are demonstrated to be inhibitors of both serotonin and norepinephrine reuptake.

Epidemiology of ovine Campylobacter infection determined by numerical analysis of electrophoretic protein profiles.

The relationship of 50 Campylobacter strains isolated from aborted ovine foetuses, and the faeces of sheep, cattle and chickens were determined by numerical analysis of electrophoretic (SDS-PAGE) protein profiles. Comparison of protein patterns by numerical methods revealed differences between C. fetus ssp. fetus, C. jejuni, and C. coli strains as well as heterogeneity among isolates from different outbreaks. Isolates from each farm produced a distinct cluster and flocks from different locations were found to be infected with relatively different strains. In most cases, protein patterns of ovine foetal isolates were very similar to those of ovine faecal isolates. Ovine isolates of C. fetus ssp. fetus, C. jejuni and C. coli gave similar protein patterns to the corresponding Campylobacter species isolated from cattle or chicken, on the same farm. Thus, it was concluded that certain protein types of ovine Campylobacter strains were more likely associated with local areas, and Campylobacter strains causing ovine abortions are distributed in the environment more widely than assumed to date.

Serotypes and electrophoretic protein profiles of Pasteurella haemolytica isolated from pneumonic ovine lungs.

Serotypes and SDS-PAGE protein profiles of P. haemolytica isolated from pneumonic ovine lungs were investigated. Of 268 P. haemolytica isolates, 232 (86.6%) were serotypable. A total of 12 serotypes were recognized in 20 different geographic origins of central Turkey. The most common serotype was A2, followed by A7, A1 and T4. Serotypes A13, A14, A16 and T15 could not be detected. In SDS-PAGE, marked differences between major bands of biotype A and T strains were found. In numerical analysis of protein profiles, biotype A and T strains were separated at 58% similarity level. Biotype A isolates produced a cluster at 80% similarity level, and biotype T isolates at 92% similarity level. No single cut off level was able to discriminate between each serotype studied and isolates could not be clustered on the basis of their geographic origins.

Motile aeromonads in the feces and carcasses of broiler chickens in turkey.

Motile Aeromonas spp. were surveyed in chicken feces, carcasses, scalding water, and chilling water during slaughtering in a commercial processing plant in Turkey. Fecal and carcass samples of 351 chickens from 15 different flocks were examined by selective isolation techniques. All of the 15 flocks were positive for motile aeromonads. Motile aeromonads were detected in 14.8%, 90.5%, and 100% of fecal, carcass, and chilling water samples, respectively, while scalding water samples were negative for these organisms. Aeromonas hydrophila was the predominant species in both feces (51.9%) and carcass (66.9%) samples, followed by A. caviae and A. sobria. These results suggested that during the slaughtering process the spread of motile aeromonads from the intestinal contents to carcasses via processing water caused a heavy contamination of chicken carcasses.

In vitro permeation of human chorioamniotic membranes by Campylobacter jejuni.

In vitro penetration of human chorioamniotic membranes by Campylobacter jejuni was investigated by an organ culture model. Membrane permeation was detected by an immunoperoxidase technique and viable bacterial counts of membrane homogenates. Human clinical isolates of C. jejuni inoculated on the maternal side of the membranes penetrated to the fetal side suggesting that chorioamniotic membranes constituted a weak barrier against Campylobacter infection. Chicken fecal isolates did not penetrate chorioamniotic membranes. In vitro culture conditions did not affect the viability of membranes. Human placental extracts and amniotic fluids enhanced the in vitro growth of C. jejuni. These results suggest that certain strains of C. jejuni may penetrate intact fetal membranes and this event may play a role in the pathogenesis of infection.

Detection of Mycoplasma ovipneumoniae and Pasteurella haemolytica antigens by an immunoperoxidase technique in pneumonic ovine lungs.

Four hundred twenty pneumonic lungs from lambs were examined for Mycoplasma ovipneumoniae and Pasteurella haemolytica by an immunoperoxidase technique using an extravidin-biotin-peroxidase complex method in formalin-fixed, paraffin-embedded sections. Histologic examination of tissue sections revealed strong positive reactions in 60.9% and 68.3% of the lungs against M. ovipneumoniae and P. haemolytica, respectively. M. ovipneumoniae and P. haemolytica antigens were observed at the surface and/or within the epithelial cells, macrophages, leucocytes, and bronchiolar exudate. The location of M. ovipneumoniae in the cytoplasm of the epithelial cells and P. haemolytica in the neutrophils was detected immunohistochemically.

Canine gastritis associated with Helicobacter felis.

Naturally occurring gastritis associated with Helicobacter felis was detected in two dogs. Histologically, chronic changes characterized by lymphofollicular aggregation were observed. Ultrastructurally, organisms located on the mucosal surface, within gastric pits and glandular epithelial cells. H. felis was isolated and urease activity was detected in all areas of the stomachs.

Sulphate-reducing bacteria in bovine faeces.

Sulphate-reducing bacteria (SRB) were found in all of 200 bovine faeces examined. The number of SRB in bovine faeces ranged from 5 x 10(2) to 6 x 10(8) bacteria g-1. Of 50 isolates identified, all were assigned to the genus Desulfovibrio.

Evaluation of immunogenicity of Campylobacter strains isolated from ovine abortions by laboratory test systems.

A microagglutination test and in vitro serum bactericidal assay were used for testing cross-reactions and cross-bactericidal activities of 15 Campylobacter strains isolated from ovine abortions. Antisera against certain strains of C. fetus subsp. fetus and C. jejuni showed higher microagglutination titres and bactericidal activity against homologous and/or heterologous strains. In both tests, cross-reactions and bactericidal activity were higher among the strains of homologous species than among those of heterologous species. The pregnant guinea-pig model was used for testing the efficacy of experimental campylobacter vaccines. Monovalent bacterins prepared with either C. fetus subsp. fetus or C. jejuni did not produce immunity to cross-challenge with the opposite species. Guinea-pigs immunized with the bivalent bacterin were found to be immune to both groups of Campylobacter when subsequently challenged. In vaccinated guinea-pigs, their in vivo ability to overcome infection is reflected in the in vitro bactericidal capacity of homologous rabbit antiserum.

Abortion associated with Campylobacter upsaliensis.

Campylobacter upsaliensis was isolated from the blood and fetoplacental material of an 18-week-pregnant woman who had contact with a household cat. We believe this is the first report of abortion associated with C. upsaliensis infection.

Studies of the pathology and microbiology of pneumonic lungs of lambs.

The characteristics of lung lesions of slaughtered lambs and the presence of microorganisms in the lesions were investigated. Pneumonia was detected in 500 (3.6%) of 13,588 lambs. Macroscopical lesions were of atypical pneumonia and were often seen in the pars cranialis of lobus cranialis dexter. Histologically, proliferative pneumonia was detected in most cases and was often accompanied by exudative characteristics. Pasteurella haemolytica was isolated from 258 (51.6%) and Mycoplasma ovipneumoniae from 215 (43.0%) of 500 pneumonic lungs. In 131 (26.2%) cases both organisms were isolated from same samples. A close relationship was found between P. haemolytica and exudative inflammation (p < 0.05).

Antimicrobial susceptibility of Pasteurella haemolytica and Pasteurella multocida isolated from pneumonic ovine lungs.

The in vitro susceptibilities of Pasteurella haemolytica biotypes A and T and P multocida from pneumonic ovine lungs to penicillin, ampicillin, oxytetracycline, chloramphenicol, erythromycin, kanamycin, neomycin, gentamicin, streptomycin and lincomycin were determined by the disk diffusion method. All the isolates were sensitive to chloramphenicol and resistant to lincomycin. The isolates of P haemolytica biotype A were consistently more sensitive to penicillin, ampicillin, oxytetracycline, erythromycin and streptomycin than those of biotype T.

In vitro infection of human fetal membranes with Candida species.

To assess whether Candida species can penetrate intact fetal membranes under in vitro conditions, Candida albicans, Candida tropicalis, Candida guilliermondii, Candida pseudotropicalis and Candida glabrata were inoculated onto the surface of the maternal side of the membranes obtained from 4 pregnant women undergoing repeat cesarean section. After incubation under culture conditions, membranes were evaluated by histological examination. C. albicans inoculated onto the maternal side penetrated and passed to the fetal side and caused some degeneration of the structure of the membrane epithelium. The other four Candida species grew heavily on the maternal surface but did not penetrate and invade the membranes. This effect of C. albicans on fetal membranes may explain the potential mechanism in the development of Candida infections of the amniotic fluid, fetal membranes and possibly the fetus.

Human follicular fluid from pre-ovulatory follicles in patients undergoing in-vitro fertilization inhibits the in-vitro growth of gram-positive micro-organisms.

In this study, we assessed the antibacterial activity of human follicular fluid obtained from patients undergoing oocyte aspiration for in-vitro fertilization and embryo transfer. Gram-positive and Gram-negative microorganisms and Candida albicans were added to the follicular fluid and broth cultures which served as controls. Follicular fluid was shown to be inhibitory against Gram-positive microorganisms; this inhibition was probably due to lysosyme and progesterone. Growth of Gram-negative micro-organisms and C. albicans were, however, supported by follicular fluid incubation.

Reversible expression of flagella in Campylobacter spp.

The in vitro phase variation of flagella and the transition rates between flagellate and aflagellate phenotypes in Campylobacter species including C. jejuni, C. coli, C. lari (thermophilic campylobacters), C. fetus subsp. fetus, C. fetus subsp. venerealis and C. hyointestinalis were investigated. The change from the flagellate to aflagellate phenotype was detected in all of the 12 Campylobacter strains studied. When measured in a motility medium, flagellate to aflagellate transition in thermophilic campylobacters, C. fetus and C. hyointestinalis strains occurred at a rate of 1.8 x 10(-3) to 7.5 x 10(-3), 3.0 x 10(-4) to 7.8 x 10(-4) and 1.8 x 10(-5) to 7.7 x 10(-6) per cell per generation, respectively. Transition from aflagellate to flagellate phenotype occurred at a rate of 5.8 x 10(-6) to 9.3 x 10(-6) per cell per generation in thermophilic campylobacters and 1.0 x 10(-6) to 1.5 x 10(-6) in C. fetus strains. No reversion from aflagellate to flagellate phenotype could be detected in C. hyointestinalis strains. It was concluded that the ability to reversibly express flagella was inherent in the wild-type strains and the transition rates for both directions were consistent for each strain.

Colonization of infant mice with flagellar variants of Campylobacter jejuni.

The role of flagella in the colonization of the intestine by Campylobacter jejuni was investigated by challenging infant mice with two flagellated strains and their nonflagellated variants. The intestinal tracts of infant mice were regularly colonized with motile strains, but not by nonmotile variants. Colonization of mice with motile C. jejuni occurred with as few as 1000 bacteria per mouse.