A 62-year-old female presented with right upper quadrant pain. Clinical examination and ultrasound scan were consistent with gallstones and acute cholecystitis. She received 3 days of intravenous Co-amoxiclav and was discharged with 5-days of oral antibiotics with arrangements to return for an elective cholecystectomy. This was performed 5 months later which revealed an inflamed gallbladder and a localised abscess secondary to gallbladder perforation. Fluid from the gallbladder was taken which cultured Raoultella planticola, a gram-negative, nonmotile environmental bacteria (Bagley et al.(1981)). This is the first report of biliary sepsis with a primary infection by R. planticola. This patient was treated with a 5-day course of oral Co-amoxiclav and made a full recovery.

A case of Raoultella ornithinolytica bacteremia in an infant with visceral heterotaxy is reported. Physical examination was remarkable for markedly red skin flushing, not unlike that seen during histamine fish poisoning. R. ornithinolytica is a histamine-producing bacterium recently elucidated as a major cause of histamine fish poisoning. Only 2 other cases of human infection by R. ornithinolytica have been reported.

The 16S-23S rRNA gene internal transcribed spacer (ITS) regions of Klebsiella spp., including Klebsiella pneumoniae subsp. pneumoniae, Klebsiella pneumoniae subsp. ozaenae, Klebsiella pneumoniae subsp. rhinoscleromatis, Klebsiella oxytoca, Klebsiella planticola, Klebsiella terrigena, and Klebsiella ornithinolytica, were characterized, and the feasibility of using ITS sequences to discriminate Klebsiella species and subspecies was explored. A total of 336 ITS sequences from 21 representative strains and 11 clinical isolates of Klebsiella were sequenced and analyzed. Three distinct ITS types-ITS(none)(without tRNA genes), ITS(glu)[with a tRNA(Glu (UUC)) gene], and ITS(ile+ala)[with tRNA(Ile (GAU)) and tRNA(Ala (UGC)) genes]-were detected in all species except for K. pneumoniae subsp. rhinoscleromatis, which has only ITS(glu) and ITS(ile+ala). The presence of ITS(none) in Enterobacteriaceae had never been reported before. Both the length and the sequence of each ITS type are highly conserved within the species, with identity levels from 0.961 to 1.000 for ITS(none), from 0.967 to 1.000 for ITS(glu), and from 0.968 to 1.000 for ITS(ile+ala). Interspecies sequence identities range from 0.775 to 0.989 for ITS(none), from 0.798 to 0.997 for ITS(glu), and from 0.712 to 0.985 for ITS(ile+ala). Regions with significant interspecies variations but low intraspecies polymorphisms were identified; these may be targeted in the design of probes for the identification of Klebsiella to the species level. Phylogenetic analysis based on ITS regions reveals the relationships among Klebsiella species similarly to that based on 16S rRNA genes.

Opportunistic pathogens have become increasingly relevant as the causative agents of clinical disease and pathological lesions in laboratory animals. This study was conducted to evaluate the role of Klebsiella oxytoca as an opportunistic pathogen in laboratory rodents. Therefore, K. oxytoca-induced lesions were studied from 2004 to early 2006 in naturally infected rodent colonies maintained at The Jackson Laboratory (TJL), Bar Harbor, USA, the Animal Research Centre (Tierforschungszentrum, TFZ) of the University of Ulm, Germany and the Central Animal Facility (ZTM) of the Hannover Medical School, Germany. K. oxytoca infections were observed in substrains of C3H/HeJ mice, which carry the Tlr4(Lps-d) allele; in LEW.1AR1-iddm rats, the latter being prone to diabetes mellitus; in immunodeficient NMRI-Foxn1(nu) mice; and in mole voles, Ellobius lutescens. The main lesions observed were severe suppurative otitis media, urogenital tract infections and pneumonia. Bacteriological examination revealed K. oxytoca as monocultures in all cases. Clonality analysis performed on strains isolated at the ZTM and TFZ (serotyping, pulse field gel electrophoresis [PFGE], enterobacterial repetitive intergenic consensus (ERIC) polymerase chain reaction, sequencing of 16S rRNA and rpoB genes) revealed that the majority of bacteria belonged to two clones, one in each facility, expressing the capsule type K55 (ZTM) or K72 (TFZ). Two strains, one isolated at the ZTM and one at the TFZ, showed different PFGE and ERIC pattern than all other isolates and both expressed capsule type K35. In conclusion, K. oxytoca is an opportunistic pathogen capable of inducing pathological lesions in different rodent species.

A 45-year-old-male presented with severe pancreatitis. Two bacterial isolates obtained from peritoneal fluid and abdominal purulent secretion were identified to the species level by 15 biochemical tests and four supplementary tests as Raoultella planticola. Identification was confirmed by rpoB gene sequencing. R. planticola is difficult to identify in the clinical laboratory, and the clinical significance of this isolation remains uncharacterized. This is the first report of pancreatitis with a primary infection by R. planticola.

Klebsiella pneumoniae and Klebsiella oxytoca are the two most frequently encountered Klebsiella species giving rise to infections in humans, but other Klebsiella species can also be found in clinical specimens: Klebsiella ozaenae, Klebsiella rhinoscleromatis, Klebsiella terrigena, Klebsiella planticola, Klebsiella ornithinolytica, and Enterobacter aerogenes (Klebsiella mobilis). However, many of these species are indistinguishable by the conventional methods employed routinely in the clinical microbiological laboratory. Several investigators have suggested various additional tests, but as yet there is no standardized test panel for identifying all Klebsiella species and subspecies. In the present study, performed in three national Klebsiella reference laboratories, we have evaluated a test panel consisting of 18 biochemical tests on 242 strains comprising all Klebsiella species and subspecies. The test panel was designed to identify organisms preliminarily identified as belonging to the genus Klebsiella on the basis of conventional methods or automated identification systems. With the described test panel it is possible to find one or more positive test results differentiating any Klebsiella species, except Klebsiella rhinoscleromatis, from its closest relative.

Enterobacterial strains of Raoultella spp. display a penicillinase-related beta-lactam resistance pattern suggesting the presence of a chromosomal bla gene. From whole-cell DNA of Raoultella planticola strain ATCC 33531(T) and Raoultella ornithinolytica strain ATCC 31898(T), bla genes were cloned and expressed into Escherichia coli. Each gene encoded an Ambler class A beta-lactamase, named PLA-1 and ORN-1 for R. planticola and R. ornithinolytica, respectively. These beta-lactamases (291 amino acids), with the same pI value of 7.8, had a shared amino acid identity of 94%, 37 to 47% identity with the majority of the chromosome-encoded class A beta-lactamases previously described for Enterobacteriaceae, and 66 to 69% identity with the two beta-lactamases LEN-1 and SHV-1 from Klebsiella pneumoniae. However, the highest identity percentage (69 to 71%) was found with the plasmid-mediated beta-lactamase TEM-1. PLA-1, which displayed very strong hydrolytic activity against penicillins, also displayed significant hydrolytic activity against cefepime and, to a lesser extent, against cefotaxime and aztreonam, but there was no hydrolytic activity against ceftazidime. Such a substrate profile suggests that the Raoultella beta-lactamases PLA-1 and ORN-1 should be classified into the group 2be of the beta-lactamase classification of K. Bush, G. A. Jacoby, and A. A. Medeiros (Antimicrob. Agents Chemother. 39:1211-1233, 1995). The highly homologous regions upstream of the bla(PLA-1A) and bla(ORN-1A) genes comprised a nucleotide sequence identical to the -35 region and another one very close to the -10 region of the bla(LEN-1) gene. From now on, as the bla gene sequences of the most frequent Raoultella and Klebsiella species are available, the bla gene amplification method can be used to differentiate these species from each other, which the biochemical tests currently carried out in the clinical laboratory are unable to do.

Bacteria of the genus Klebsiella are important opportunistic pathogens responsible for nosocomial infections that are increasingly resistant to antimicrobial agents. Distinctive identification of the species K. oxytoca, K. pneumoniae, K. planticola, K. ornithinolytica and K. terrigena is difficult based on phenotypic tests and misidentifications are frequent in routine clinical microbiology. We developed a specific method to discriminate K. oxytoca from the other species of the genus Klebsiella, based on the PCR amplification of the polygalacturonase (pehX) gene. A PCR amplicon of 344 bp was obtained in all 35 K. oxytoca strains tested, but in none of the 29 K. pneumoniae, 12 K. planticola/K. ornithinolytica and 7 K. terrigena strains tested. The test was also negative for polygalacturonate-degrading species of the genus Erwinia. Analysis of 24 strains designated as K. pneumoniae from international collections (NCTC, PZH) revealed previous misidentification of six K. oxytoca strains. Key biochemical tests fully confirmed the pehX PCR results. The new K. oxytoca identification assay should be useful for both clinical and ecological monitoring of K. oxytoca strains, as well as for controlling the previous identification of collection strains.

Histamine fish poisoning is caused by histamine-producing bacteria (HPB). Klebsiella pneumoniae and Klebsiella oxytoca are the best-known HPB in fish. However, 22 strains of HPB from fish first identified as K. pneumoniae or K. oxytoca by commercialized systems were later correctly identified as Raoultella planticola (formerly Klebsiella planticola) by additional tests. Similarly, five strains of Raoultella ornithinolytica (formerly Klebsiella ornithinolytica) were isolated from fish as new HPB. R. planticola and R. ornithinolytica strains were equal in their histamine-producing capabilities and were determined to possess the hdc genes, encoding histidine decarboxylase. On the other hand, a collection of 61 strains of K. pneumoniae and 18 strains of K. oxytoca produced no histamine.

The new VITEK 2 system (bioMérieux) was evaluated at two independent sites with the identification card for gram-negative bacilli (ID-GNB card). Of the 845 strains tested, which represented 70 different taxa belonging to either the family Enterobacteriaceae or the nonenteric bacilli, 716 (84.7%) were correctly identified at the species level. Thirty-two (3.8%) additional strains were identified to the species level after the performance of simple, rapid manual tests (oxidase, hemolysis, indole reaction, motility, and pigmentation). For 80 (9.5%) strains, these additional tests did not lead to an identification at the species level but the correct species identification was given among the organisms listed. Only 7 (0.8%) strains were misidentified, and 10 (1.2%) were not identified. Mistakes were randomly distributed over different taxa. Due to the new, more sensitive fluorescence-based technology of the VITEK 2 system, final results were available after 3 h. Since our evaluation was mainly a stress test, it is predicted that the VITEK 2 system in conjunction with the ID-GNB card would perform well under conditions of a routine clinical laboratory in identifying members of the family Enterobacteriaceae and selected species of nonenteric bacteria. This system is a promising, highly automated new tool for the rapid identification of gram-negative bacilli from human clinical specimens.

A semi-automated commercial system (ID 32 E, bioMérieux) for 24-hour identification of Enterobacteriaceae and other gram-negative fermentative and nonfermentative bacteria encountered in diagnostic microbiology was evaluated. Overall, the system correctly identified 506 (91.5%) of the 553 strains tested, 94 (17.0%) strains requiring additional tests for complete identification. Six (1.1%) strains were misidentified and 33 (6.0%) strains were not identified. Eight (1.4%) strains were not present in the database and were misidentified or not identified. The system is a suitable alternative to existing systems for the identification of Enterobacteriaceae and other gram-negative bacteria frequently encountered in clinical samples.

Two hundred and four strains of Gram-negative bacteria of clinical origin, initially identified as Klebsiella using the API 20 E system, and 10 reference strains were further analysed with the API 20 EC test system and the API 50 CH, API 50 AO, API 50 AA assimilation systems. Four clusters corresponding to the species Klebsiella pneumoniae subsp. pneumoniae, K. oxytoca, K. planticola, and K. terrigena were formed after numerical analysis of 155 selected tests and the 26 most discriminating tests were determined. A comparison was made between conventional identification using the API 20 E system and the results of the numerical analysis. The conventional method resulted in incorrect identification of 13% of the strains tested, especially for the new species: K. planticola and K. terrigena. After numerical analysis, 17 out of 204 strains (8.3%) of clinical origin were identified as K. planticola. Only 1 strain of clinical origin was identified as K. terrigena, and 1 strain as K. ornithinolytica.

The spread of multiresistant bacteria increases the need for new antibiotics. The observation that some nucleoside analogues have antibacterial activity led us to further investigate the antimicrobial activity and resistance of zidovudine (AZT). We determined the minimum inhibition concentration (MIC), studied time-kill curves, induced resistant bacteria and sequenced the gene for thymidine kinase. We demonstrate that AZT has a bactericidal effect on some enterobacteria. However, AZT could induce resistance in Escherichia coli. These resistances were associated with various modifications in the thymidine kinase gene. In particular, we observed the presence in this gene of an insertion sequence (IS) similar to IS911 of Shigella dysenteriae in two resistant clones. No cross-resistance with classical antibiotics in strains with modified thymidine kinase gene was observed. Finally, an additive or synergistic activity between AZT and the two aminoglycoside antibiotics amikacin and gentamicin was observed. We demonstrate the bactericidal activity of AZT and show synergy in association with gentamicin. Genetic modifications in resistant bacteria were identified. Our results indicate that AZT could potentially be added in the treatment of infections with enterobacteria or represent the basis for the development of derivatives with better activity and inducing less resistance.

This study reports biochemical composition and morphological aspect of gallstones as investigated by spectroscopy IR method. Participants were 24 patients composed of 12 males and 12 females who underwent cholecystectomy with age mean of 44.8 years. The gallstones were classified either as pigments stones (n = 12), cholesterol stones (n = 8) or as mixed stones (n = 4) according to analysis by Fourier transform infrared spectroscopy. The infrared spectroscopy quantification reported eight stones contained 100% of cholesterol, eight of 100% of calcium bilirubinate, four stones were composed of 65% calcium bilirubinate phosphate and 35% calcium carbonate, and four stones contained 65% cholesterol, 30% neutral calcium bilirubinate, 5% protein and traces of calcium bilirubinate acid. Our findings showed that most gallstones were composed of pigment stones with relative large proportion of cholesterol stones, whereas previous study in Caucasian reported predominance of cholesterol stones. These findings indicate the influence of diet and chronic haemolysis in the stones formation in regard to biochemical composition differences between those found in European area and our results. Therefore, Fourier transform infrared spectroscopy method allowed to determine quality and quantity of biochemical components of gallstones. Therefore, a careful survey must allow knowing the nutritional and environmental factors in the occurrence of gallstones to Côte d'Ivoire, in order to prevent this disease.

Cataract surgery is a usually successful procedure that restores vision by replacing the natural lens with an intraocular lens (IOL). Acute postoperative endophthalmitis is still one of the most serious complications of cataract surgery. Its incidence has been reported to be between 0.04% and 0.32%. Precisely why bacteria induce endophthalmitis is not entirely understood. Indeed the risk of its development may be influenced by several factors. Among them, bacterial adhesion to the IOL has been recently emphasized in the ophthalmology literature. Indeed, the ability of an organism to adhere to the IOL surface is believed to be associated with a risk of infection at the implantation site. Several studies have demonstrated that bacterial adhesion is influenced by IOL materials. Ever since, numerous studies have investigated the interactions between bacteria and different types of IOLs to determine which biomaterial would be most permissive to bacterial adherence. This article reviews all the epidemiological and experimental data relating to the study of the relationship between bacterial adhesion, IOL material, and risk of developing postoperative endophthalmitis. Even if discrepancies between these studies exist, mainly stemming from the use of different experimental conditions and protocols, it seems that bacterial adhesion is strongly influenced by IOL material. Epidemiological studies suggest that the implantation of silicone IOLs might be associated with increased rates of endophthalmitis. Experimental studies reach similar conclusions showing that hydrophobic IOLs such as silicone or acrylic hydrophobic IOLs are more permissive to bacterial adhesion and growth than hydrophilic IOLs such as acrylic hydrophilic IOLs. Among the interactions that govern bacterial attachment to the IOLs, it seems that hydrophilic-hydrophobic interactions have the greatest influence. Nevertheless, since bacterial adhesion is a complicated process affected by many factors, the conclusions drawn by these results have to be interpreted with care. Further investigations are still needed to understand the connections between IOL material and endophthalmitis.

The purpose of this study was to evaluate the possibility of using a semi-automated repetitive DNA sequences-based polymerase chain reaction (rep-PCR) for typing Pseudomonas aeruginosa isolates. rep-PCR profiles obtained by the DiversiLab(R) system of 84 P. aeruginosa isolates from distinct epidemiological situations were obtained. rep-PCR groupings were in good agreement with the origin of these isolates. Linked rep-PCR profiles were observed for isolates recovered from a same family of cystic fibrosis (CF) patients, for the etiological agents of clustered cases of nosocomial infections, and for some isolates recovered from a same hospital room. rep-PCR and pulsed-field gel electrophoresis SpeI restricted genomic DNA (PFGE-SpeI) profiles were compared. In a few instances, rep-PCR revealed genetic divergences among isolates of a same group of PFGE-SpeI profiles. These divergences could reflect genetic drifts among closely related isolates, as illustrated by those observed between clinical and environmental isolates of a same group of PFGE-SpeI profiles. The interpretation of such differences will require further studies, but the rep-PCR analysis of P. aeruginosa diversity appeared to be an appropriate method to investigate infra-specific genetic relatedness.

This study aimed at describing the levels of use of oseltamivir in 12 European Union (EU) Member States and European Economic Area (EEA)/European Free Trade Area (EFTA) countries. The data were converted into prescription rates and compared with the national proportions of resistant influenza A(H1N1) viruses through regression analysis.

Efflux pumps located in the bacterial membranes are responsible for low level resistance to antibiotics, considered not to be relevant in the clinic and thus often neglected. However, these pumps contribute to the emergence of high level antibiotic resistance mechanisms, which are responsible for severe complications during the treatment of infectious diseases. Therefore it is necessary to take into account these pumps while developing novel antibacterial agents. Among these new research strategies, the development of efflux pump inhibitors seems to be an attractive approach to restore the activity of some "classical" antibiotics and to limit the emergence of multiresistant strains associated with hospital-acquired infections. In this review, we focalise on Staphylococcus aureus efflux pumps and their potential inhibitors.

Clinical signs of malaria are the combined expression of several biological mechanisms. During this parasite infection, anaemia can be the consequence of several different pathogenic mechanisms. It can be an acute haemolytic anaemia due to a mechanical and immune action of the parasite or an inflammation. Besides, in Africa malaria matches with iron deficiency area. So, malarial anaemia in tropical area can be a characteristic of iron deficiency The purpose of this survey was to define the features of malarial anaemia and elucidate the link of all biological processes involved. A black population living in tropical urban areas, with fever and diagnosed Plasmodium-infection was assessed. Parasitaemia, haemoglobin, hematocrit, average corpuscular volume and average corpuscular haemoglobin were determined. For each patient, iron index status and acute phase protein were assessed with the plasmatic iron, ferritin, haptoglobin, transferrin and C-reactive protein. Regardless of gender and age, the characteristics of malarial anaemia are microcythaemia and hypochromia. Anaemia occurs as frequently as parasitaemia is high. When parasitaemia is low anaemia gets a haemolytic feature. When parasitaemia is high, anaemia gets haemolytic and inflammatory features. Anaemia occurs more often with a good iron index status.

Four phenolic acids, namely 2-[(Z)-heptadec-11-enyl]-6-hydroxybenzoic acid (1), 2-[(6Z,9Z,12Z)-heptadeca-6,9,12-trienyl]-6-hydroxybenzoic acid (2), 2-[(9Z,12Z)-heptadeca-9,12-dienyl]-6-hydroxybenzoic acid (3), and 2-hydroxy-6-(12-phenyldodecyl)benzoic acid (4), and one sesquiterpene, asperpenoid (5), were isolated from the 95% EtOH extract of the roots of Homalomena occulta, among which 1, 2, and 5 represent new compounds. Further, the phenolic acids 1-4 exhibited BACE1 (beta-secretase) inhibitory activity with IC(50) values of 6.23+/-0.94, 6.28+/-0.63, 7.93+/-0.38, and 7.65+/-0.62 muM, respectively.

One hundred four Enterobacter isolates were tested by standard CLSI disk diffusion methods for detecting extended-spectrum beta-lactamases (ESBLs) and with cefepime-clavulanate disk combinations. SHV-12 was produced by 8.7% of isolates. The cefepime-clavulanate combination provided 88% sensitivity and 91% specificity for the detection of SHV-12 ESBL.

Two new butenolide derivatives,(4R,7R*/S*)-2, 4-dimethyl-4-(1'-acetyl-3'-oxobutyl)-2- butenolide and its epimer,(4R,7S*/R*)-2, 4-dimethyl-4-(1'-acetyl-3'-oxobutyl)-2- butenolide were isolated from the acetone extract of the whole plant of Saussurea katochaete. Their structures were established by various spectral methods including IR, 1D-, 2D-NMR and HR-ESIMS. Four known compounds were also isolated and identified as 2, 4-dimethyl-4-hydroxy-2- butenolide, 3-hexene-2, 5-dione, 3-hydroxyhexane-2, 5-dione, 1-hydroxy-(Z)-3-hexene-2, 5-dione.

In leaves, roots and calli of P. dodecandra we found two 28,30-dicarboxyolean-12-enes and their glycosides in addition to the hitherto known four 28-monocarboxyolean-12-enes on which the chemotaxonomy was based. Therefore, P. dodecandra can no longer be considered as chemotaxonomically different from the other Phytolacca species.

A plasmid-encoded class II transposon element was identified in a carbapenem-resistant Pseudomonas putida isolate. Tn1332, closely related to Tn1331, harbored the metallo-beta-lactamase gene bla(VIM-2) in addition to four other antibiotic resistance genes, aacA4, aadA1, bla(OXA-9), and bla(TEM-1), and two novel insertion sequences, ISPpu17 and ISPpu18.

Four putative type IV pilus genes from the toxic, naturally transformable Microcystis aeruginosa PCC7806 were identified. Three of these genes were clustered in an arrangement which is identical to that from other cyanobacterial genomes. Type IV pilus-like appendages were also observed by electron microscopy.

Five fungal species were isolated and identified in food products: Ascotricha chartarum, Leptosphaerulina argentinensis, Veronaea coprophila, Scolecobasidium constrictum and Coremiella cubispora. A. chartarum was isolated from paper bag containing sugar and the other four from tomato sauce. Except for L. argentinensis, the other four were new reports and the five species were isolated for the first time in Argentina.

The genus Saccharum consists of two wild and four cultivated species. Novel interspersed sequences were isolated from cultivated sugar cane S. officinarum. These sequences were accumulated in all four cultivated species and their wild ancestral species S. robustum, but were not detected in the other wild species S. spontaneum and the relative Erianthus arundinaceus. The species-specific accumulation of interspersed sequences would correlate to the domestication of sugar canes.

Four new eudesmanolides and two new guaianolides were isolated from the aerial parts of Anthemis carpatica Willd. and their structures elucidated by spectral methods. In addition, seven known sesquiterpene lactones were identified.

Two new steroids, 3 beta-vanilloyloxy-stigmast-5-ene-7-one, and stigmast-5-ene-3 beta,7 beta,15 alpha-triol were isolated from the roots of Adenophora stenanthina subsp. xifengensis along with four known compounds, stigmast-5-ene-3 beta-ol-7-one, stigmast-5-ene-3 beta,7 beta-diol, syringin, sinapyl alcohol 1, 3'-di-O-beta-D-glucopyranoside. Their structures were elucidated on the basis of spectral methods.