Plautus, Menaechmi: Twin Helping Twin

This paper explores how Menaechmus II of Syracuse unintentionally succeeds in removing Menaechmus I, his Epidamnian twin, from a society which has been exploiting him. The process of withdrawal runs through the play, and is achieved in two stages in which Menaechmus II assumes his twin brother’s meta-dramatic role:  in the first play the newly arrived brother alienates the Epidamnian twin from his immediate social and family milieu; and in the second he almost leads the Epidamnians to have his brother isolated within the community on account of the latter’s alleged insanity. As servus bonus, Messenio initially discourages Menaechmus II from comic merrymaking, thus impeding the reunion of the siblings, but later assists in forwarding the plot: he thwarts the Epidmanians’ intention to seclude him and eventually facilitates the recognitio between the twins as well as their final decision to return to their native land. Thus, Menaechmus II’s quest for his twin, which seems to have been deferred when he first arrives in Epidamnus, is prepared for and effectively carried through via the evolution of meta-plots

High throughput detection of Coxiella burnetii by real-time PCR with internal control system and automated DNA preparation

<p>Abstract</p> <p>Background</p> <p><it>Coxiella burnetii </it>is the causative agent of Q-fever, a widespread zoonosis. Due to its high environmental stability and infectivity it is regarded as a category B biological weapon agent. In domestic animals infection remains either asymptomatic or presents as infertility or abortion. Clinical presentation in humans can range from mild flu-like illness to acute pneumonia and hepatitis. Endocarditis represents the most common form of chronic Q-fever. In humans serology is the gold standard for diagnosis but is inadequate for early case detection. In order to serve as a diagnostic tool in an eventual biological weapon attack or in local epidemics we developed a real-time 5'nuclease based PCR assay with an internal control system. To facilitate high-throughput an automated extraction procedure was evaluated.</p> <p>Results</p> <p>To determine the minimum number of copies that are detectable at 95% chance probit analysis was used. Limit of detection in blood was 2,881 copies/ml [95%CI, 2,188–4,745 copies/ml] with a manual extraction procedure and 4,235 copies/ml [95%CI, 3,143–7,428 copies/ml] with a fully automated extraction procedure, respectively. To demonstrate clinical application a total of 72 specimens of animal origin were compared with respect to manual and automated extraction. A strong correlation between both methods was observed rendering both methods suitable. Testing of 247 follow up specimens of animal origin from a local Q-fever epidemic rendered real-time PCR more sensitive than conventional PCR.</p> <p>Conclusion</p> <p>A sensitive and thoroughly evaluated real-time PCR was established. Its high-throughput mode may show a useful approach to rapidly screen samples in local outbreaks for other organisms relevant for humans or animals. Compared to a conventional PCR assay sensitivity of real-time PCR was higher after testing samples from a local Q-fever outbreak.</p

Molecular characterization of Coxiella burnetii isolates by infrequent restriction site-PCR and MLVA typing

BACKGROUND: Coxiella burnetii, the causative agent of Q fever, has a wide host range. Few epidemiological tools are available, and they are often expensive or not easily standardized across laboratories. In this work, C. burnetii isolates from livestock and ticks were typed using infrequent restriction site-PCR (IRS-PCR) and multiple loci variable number of tandem repeats (VNTR) analysis (MLVA). RESULTS: By applying IRS-PCR, 14 C. burnetii isolates could be divided into six groups containing up to five different isolates. Clustering as deduced from MLVA typing with 17 markers provided an increased resolution with an excellent agreement to IRS-PCR, and with the plasmid type of each strain. MLVA was then applied to 28 additional C. burnetii isolates of different origin and 36 different genotypes were identified among the 42 isolates investigated. The clustering obtained is in agreement with published Multiple Locus Sequence Typing (MLST) data. Two panels of markers are proposed, panel 1 which can be confidently typed on agarose gel at a lower cost and in any laboratory setting (10 minisatellite markers with a repeat unit larger than 9 bp), and panel 2 which comprises 7 microsatellites and provides a higher discriminatory power. CONCLUSION: Our analyses demonstrate that MLVA is a powerful and promising molecular typing tool with a high resolution and of low costs. The consistency of the results with independent methods suggests that MLVA can be applied for epidemiological studies. The resulting data can be queried on a dedicated MLVA genotyping Web service

Current perspectives on the transmission of Q fever: Highlighting the need for a systematic molecular approach for a neglected disease in Africa.

Q fever is a bacterial worldwide zoonosis (except New Zealand) caused by the Gram-negative obligate intracellular bacterium Coxiella burnetii (C. burnetii). The bacterium has a large host range including arthropods, wildlife and companion animals and is frequently identified in human and livestock populations. In humans, the disease can occur as either a clinically acute or chronic aetiology, affecting mainly the lungs and liver in the acute disease, and heart valves when chronic. In livestock, Q fever is mainly asymptomatic; however, the infection can cause abortion, and the organism is shed in large quantities, where it can infect other livestock and humans. The presence of Q fever in Africa has been known for over 60 years, however while our knowledge of the transmission routes and risk of disease have been well established in many parts of the world, there is a significant paucity of knowledge across the African continent, where it remains a neglected zoonosis. Our limited knowledge of the disease across the African sub-continent have relied largely upon observational (sero) prevalence studies with limited focus on the molecular epidemiology of the disease. This review highlights the need for systematic studies to understand the routes of C. burnetii infection, and understand the disease burden and risk factors for clinical Q fever in both humans and livestock. With such knowledge gaps filled, the African continent could stand a better chance of eradicating Q fever through formulation and implementation of effective public health interventions

Cell-Free Propagation of Coxiella burnetii Does Not Affect Its Relative Virulence

Q fever is caused by the obligate intracellular bacterium Coxiella burnetii. In vitro growth of the bacterium is usually limited to viable eukaryotic host cells imposing experimental constraints for molecular studies, such as the identification and characterisation of major virulence factors. Studies of pathogenicity may benefit from the recent development of an extracellular growth medium for C. burnetii. However, it is crucial to investigate the consistency of the virulence phenotype of strains propagated by the two fundamentally different culturing systems. In the present study, we assessed the viability of C. burnetii and the lipopolysaccaride (LPS) encoding region of the bacteria in both culture systems as indirect but key parameters to the infection potential of C. burnetii. Propidium monoazide (PMA) treatment-based real-time PCR was used for enumeration of viable C. burnetii which were validated by fluorescent infectious focus forming unit counting assays. Furthermore, RNA isolated from C. burnetiipropagated in both the culture systems was examined for LPS-related gene expression. All thus far known LPS-related genes were found to be expressed in early passages in both culturing systems indicating the presence of predominantly the phase I form of C. burnetii. Finally, we used immune-competent mice to provide direct evidence, that the relative virulence of different C. burnetii strains is essentially the same for both axenic and cell-based methods of propagation

Q fever in pregnant Goats: PAthogenesis and excretion of Coxiella burnetii

Coxiella burnetii is an intracellular bacterial pathogen that causes Q fever. Infected pregnant goats are a major source of human infection. However, the tissue dissemination and excretion pathway of the pathogen in goats are still poorly understood. To better understand Q fever pathogenesis, we inoculated groups of pregnant goats via the intranasal route with a recent Dutch outbreak C. burnetii isolate. Tissue dissemination and excretion of the pathogen were followed for up to 95 days after parturition. Goats were successfully infected via the intranasal route. PCR and immunohistochemistry showed strong tropism of C. burnetii towards the placenta at two to four weeks after inoculation. Bacterial replication seemed to occur predominantly in the trophoblasts of the placenta and not in other organs of goats and kids. The amount of C. burnetii DNA in the organs of goats and kids increased towards parturition. After parturition it decreased to undetectable levels: after 81 days post-parturition in goats and after 28 days post-parturition in kids. Infected goats gave birth to live or dead kids. High numbers of C. burnetii were excreted during abortion, but also during parturition of liveborn kids. C. burnetii was not detected in faeces or vaginal mucus before parturition. Our results are the first to demonstrate that pregnant goats can be infected via the intranasal route. C. burnetii has a strong tropism for the trophoblasts of the placenta and is not excreted before parturition; pathogen excretion occurs during birth of dead as well as healthy animals. Besides abortions, normal deliveries in C. burnetii-infected goats should be considered as a major zoonotic risk for Q fever in humans

Immunologic Response of Unvaccinated Workers Exposed to Anthrax, Belgium

To determine immunologic reactivity to Bacillus anthrax antigens, we conducted serologic testing of workers in a factory that performed scouring of wool and goat hair. Of 66 workers, ≈10% had circulating antibodies or T lymphocytes that reacted with anthrax protective antigen. Individual immunity varied from undetectable to high

Coxiella burnetii (Q fever) seroprevalence in prey and predators in the United kingdom: evaluation of infection in wild rodents, foxes and domestic cats using a modified ELISA

Coxiella burnetii, the agent of Q-fever, is recognized as a worldwide zoonosis with a wide host range and potentially complex reservoir systems. Infected ruminants are the main source of infection for humans, but cats and other mammals, including wild rodents, also represent potential sources of infection. There has been a recent upsurge of reported cases in humans, domestic ruminants and wildlife in many parts of the world, and studies have indicated that wild brown rats may act as true reservoirs for C. burnetii and be implicated in outbreaks in livestock and humans. However, investigation of reservoir systems is limited by lack of validated serological tests for wildlife or other non-target species. In this study, serum samples from 796 wild rodents (180 bank voles, 309 field voles, 307 wood mice) 102 wild foxes and 26 domestic cats from three study areas in the UK were tested for the presence of antibodies to C. burnetii using a commercial indirect ELISA kit modified for use in multiple wildlife species. Test thresholds were determined for each species in the absence of species-specific reference sera using a bi-modal latent class mixture model to discriminate between positive from negative results. Based on the thresholds determined, seroprevalence in the wild rodents ranged from 15.6% to 19.1% depending on species (overall 17.3%) and was significantly higher in both foxes (41.2%) and cats (61.5%) than in rodents. This is the first report to quantify seroprevalence to C. burnetii in bank voles, field voles, wood mice, foxes and cats in the UK and provides evidence that predator species could act as indicators for the presence of C. burnetii in rodents. The study demonstrates that wildlife species could be significant reservoirs of infection for both livestock and humans, and the high seroprevalence in domestic cats highlights the potential zoonotic risk from this species