Identification and Antimicrobial Resistance of Campylobacter Species Isolated from Animal Sources

Background: Campylobacter spp. are together with Salmonella spp. the leading causes of human bacterial gastroenteritis worldwide. The most commonly isolated species in humans are Campylobacter jejuni and C. coli. The isolation, identification, and antimicrobial resistance of Campylobacter spp. from poultry and raw meat from slaughterhouses, has been investigated for the first time in Greece. During the period from August 2005 to November 2008 a total of 1080 samples were collected: (a) 830 fecal samples from five poultry farms, (b) 150 cecal samples from chicken carcasses in a slaughterhouse, and (c) 100 fecal samples from one pig farm near the region of Attica. The identification of the isolates was performed with conventional (sodium hippurate hydrolysis and commercial identification system (Api CAMPY system, bioMerieux, France), as well as with and molecular methods based on 16S rRNA species specific gene amplification by PCR and subsequent sequence analysis of the PCR products. Results: Sixteen Campylobacter strains were isolated, all collected from the poultry farms. None of the strains was identified as C. jejuni. Antimicrobial susceptibility to six antimicrobials was performed and all the strains were susceptible to ciprofloxacin, amoxicillin–clavulanic acid, and gentamicin. Thirteen out of 14 C. coli were resistant to erythromycin and all C. coli strains were resistant to ampicillin. Conclusion: Our results emphasize the need for a surveillance and monitoring system with respect to the prevalence and antimicrobial resistance of Campylobacter in poultry, as well as for the use of antimicrobials in veterinary medicine in Greece

Incidence and antimicrobial susceptibilities of genital mycoplasmas in outpatient women with clinical vaginitis in Athens, Greece

Objectives: The incidence and antimicrobial susceptibilities of Ureaplasma urealyticum and Mycoplasma hominis, isolated from vaginal and endocervical swabs collected from 369 outpatient women, were determined. Methods: Isolation, identification and typing of the pathogens were performed by means of conventional methods. The antimicrobial susceptibilities of the genital mycoplasmas were determined with commercially available kits and evaluated according to the CLSI. Results and conclusions: In 65 (47.44%) out of the 137 positive specimens, U. urealyticum was grown as a single pathogen, in 0.72% M. hominis was grown as a single pathogen and in 2.92% both urogenital mycoplasmas were grown. In the remaining specimens (48.90%), there was a mixed growth with other microbes. Of the isolated U. urealyticum strains, 87.4% and 98.2% were susceptible to tetracycline and doxycycline, respectively, 79.2% were susceptible to josamycin, 48.6% were susceptible to clarithromycin and 91.8% were susceptible to pristinamycin, while erythromycin, azithromycin, ciprofloxacin and ofloxacin proved to be inactive against most of the strains. M. hominis isolates were 100% susceptible to tetracycline, doxycycline and pristinamycin, while susceptibilities to the other antimicrobial agents varied mainly in the range of ‘intermediate’ or ‘resistant’. As results originating from similar studies from various countries are very controversial, the simplest way to avoid therapeutic failures would be the implementation of rational treatment regimens based on culture isolation and the in vitro determination of the antimicrobial susceptibility of genital mycoplasmas in each clinical case

The Telomere/Telomerase System in Chronic Inflammatory Diseases. Cause or Effect?

Telomeres are specialized nucleoprotein structures located at the end of linear chromosomes and telomerase is the enzyme responsible for telomere elongation. Telomerase activity is a key component of many cancer cells responsible for rapid cell division but it has also been found by many laboratories around the world that telomere/telomerase biology is dysfunctional in many other chronic conditions as well. These conditions are characterized by chronic inflammation, a situation mostly overlooked by physicians regarding patient treatment. Among others, these conditions include diabetes, renal failure, chronic obstructive pulmonary disease, etc. Since researchers have in many cases identified the association between telomerase and inflammation but there are still many missing links regarding this correlation, the latest findings about this phenomenon will be discussed by reviewing the literature. Our focus will be describing telomere/telomerase status in chronic diseases under the prism of inflammation, reporting molecular findings where available and proposing possible future approaches

The Telomere/Telomerase System in Chronic Inflammatory Diseases. Cause or Effect?

Telomeres are specialized nucleoprotein structures located at the end of linear chromosomes and telomerase is the enzyme responsible for telomere elongation. Telomerase activity is a key component of many cancer cells responsible for rapid cell division but it has also been found by many laboratories around the world that telomere/telomerase biology is dysfunctional in many other chronic conditions as well. These conditions are characterized by chronic inflammation, a situation mostly overlooked by physicians regarding patient treatment. Among others, these conditions include diabetes, renal failure, chronic obstructive pulmonary disease, etc. Since researchers have in many cases identified the association between telomerase and inflammation but there are still many missing links regarding this correlation, the latest findings about this phenomenon will be discussed by reviewing the literature. Our focus will be describing telomere/telomerase status in chronic diseases under the prism of inflammation, reporting molecular findings where available and proposing possible future approaches

Inquiring into the Gaps of Campylobacter Surveillance Methods

Campylobacter is one of the most common pathogen-related causes of diarrheal illnesses globally and has been recognized as a significant factor of human disease for more than three decades. Molecular typing techniques and their combinations have allowed for species identification among members of the Campylobacter genus with good resolution, but the same tools usually fail to proceed to subtyping of closely related species due to high sequence similarity. This problem is exacerbated by the demanding conditions for isolation and detection from the human, animal or water samples as well as due to the difficulties during laboratory maintenance and long-term storage of the isolates. In an effort to define the ideal typing tool, we underline the strengths and limitations of the typing methodologies currently used to map the broad epidemiologic profile of campylobacteriosis in public health and outbreak investigations. The application of both the old and the new molecular typing tools is discussed and an indirect comparison is presented among the preferred techniques used in current research methodolog

The first database comprised of flagellin gene (flaA) types of Campylobacter jejuni human clinical isolates from Greece

Flagellin subunit A gene (flaA) typing of Campylobacter has been recognized by several groups as a relatively simple and quick genotyping method. The present study aimed to create, for the first time in Greece, a database with flaA restriction patterns, which could be used for future epidemiological and clinical studies. A total of 207 C. jejuni clinical isolates of known serotype were collected from 5 general hospitals of the area of Attica, during the period 2000-2003. The RFLP profiles of each strain were matched in 44 bins of 0 or 1. Thirty nine different flaA types, designated as flaA 1 GR to flaA 39 GR (GR: Greece) were found. There was no significant association of certain genotypes with certain serotypes. However flaA typing showed a remarkable discriminatory ability inside the non-typable (NT) group. Evaluating our results we observed (i) that there was no clonality of a certain flaA type among the strains and the serotypes examined and (ii) that the discriminatory ability of flaA typing was much better than that of serotyping. Giving a simple and detailed description of the data analysis, we are the first who publish the bin patterns for the flaA genotypes found

Acute Bacterial Meningitis Cases Diagnosed by Culture and PCR in a Children's Hospital Throughout a 9-Year Period (2000-2008) in Athens, Greece

Background and Objectives: Acute bacterial meningitis is one of the most severe infectious diseases, affecting mainly infants and, secondarily, older children and adolescents. Diagnosis in the early stages is often difficult and despite treatment with appropriate antibiotic therapy, the case fatality rate remains high. In the present study, the incidence of bacterial meningitis was registered in a general pediatric hospital in Athens, Greece, during a 9-year period (2000-2008), and the use of molecular methods in the diagnosis of bacterial meningitis versus the conventional cultural methods was evaluated. The impact of vaccination against meningitis-causing bacteria on the incidence of bacterial meningitis was also assessed. Methods: From a total of 1833 children hospitalized with suspected clinical symptoms and signs of meningitis, all cerebrospinal fluid (CSF) and blood samples were analyzed by white blood cell (WBC) count, measurement of glucose, protein, and C-reactive protein (CRP) levels, as well as by conventional bacteriologic culture methods. If samples showed altered CSF markers that were consistent with meningitis in general, they were further investigated by PCR for bacterial pathogens. Results: Of the 1833 patients, 289 (15.76%) were found to be positive for meningitis after CSF examination, based on white blood cell count and differentiation, glucose, protein, and CRP. Fifty-six of the 289 (19.37%) had confirmed bacterial meningitis, as diagnosed by either culture and/or PCR. Of these 56 cases, 44(78.6%) were detected only by PCR, and 12 cases (21.4%) were confirmed by PCR and culture. The predominant microorganism was Neisseria meningitidis serogroup B (n = 40; 71.4%), followed by Streptococcus pneumoniae not typed [NT] (n = 7; 12.5%), Streptococcus spp. (n = 4; 7.1%), Haemophilus influenzae NT (n = 2; 3.6%), and S. pneumoniae serotype 3, Streptococcus group B, and S. pneumoniae serotype 18C (each n = 1; 1.8%). Conclusion: In Greece, according to data from the National Meningitis Reference Laboratory, vaccination against N. meningitidis serogroup C since 2001 led to a 10-fold decrease in the incidence of meningitis cases, vaccination against S. pneumoniae serotypes included in the heptavalent conjugate vaccine since 2005 led to a 3.4-fold incidence decrease, and vaccination against H. influenzae type b since 1992 led almost to an absence of cases. In the population of the present study, none of the cases were caused by the above-mentioned vaccine pathogens, except for one S. pneumoniae serotype 18C case with no history of past vaccination. The introduction of vaccination against meningitis-causing bacteria has drastically decreased the emergence of the infection. The improved molecular amplification assays proved to be superior to conventional bacteriologic methods and should be introduced into routine diagnosis, as well as the epidemiologic surveillance of bacterial meningitis