Overleving van Coxiella burnetii in geitenmest : eindrapportage

De afname van het aantal bacteriën in geitenmest is bestudeerd onder omstandigheden, zoals die zich in opgeslagen geitenmest voordoen. De afname van het aantal bacteriën bij verschillende temperaturen wordt weergegeven door middel van de decimale reductietijd (DRT of Dwaarde). De DRT is de tijd die nodig is om het aantal bacteriën met een factor 10 te laten afnemen bij een bepaalde temperatuur. In dit onderzoek is de DRT voor C. burnetii in geitenpotstalmest bij verschillende temperaturen bepaald

Detection of Coxiella burnetii in complex matrices by using multiplex quantitative PCR during a major Q fever outbreak in the Netherlands

Q fever, caused by Coxiella burnetii, is a zoonosis with a worldwide distribution. A large rural area in the southeast of the Netherlands was heavily affected by Q fever between 2007 and 2009. This initiated the development of a robust and internally controlled multiplex quantitative PCR (qPCR) assay for the detection of C. burnetii DNA in veterinary and environmental matrices on suspected Q fever-affected farms. The qPCR detects three C. burnetii targets (icd, com1, and IS1111) and one Bacillus thuringiensis internal control target (cry1b). Bacillus thuringiensis spores were added to samples to control both DNA extraction and PCR amplification. The performance of the qPCR assay was investigated and showed a high efficiency; a limit of detection of 13.0, 10.6, and 10.4 copies per reaction for the targets icd, com1, and IS1111, respectively; and no crossreactivity with the nontarget organisms tested. Screening for C. burnetii DNA on 29 suspected Q fever-affected farms during the Q fever epidemic in 2008 showed that swabs from dust-accumulating surfaces contained higher levels of C. burnetii DNA than vaginal swabs from goats or sheep. PCR inhibition by coextracted substances was observed in some environmental samples, and 10- or 100-fold dilutions of samples were sufficient to obtain interpretable signals for both the C. burnetii targets and the internal control. The inclusion of an internal control target and three C. burnetii targets in one multiplex qPCR assay showed that complex veterinary and environmental matrices can be screened reliably for the presence of C. burnetii DNA during an outbreak. © 2011, American Society for Microbiology

Polycystic ovary syndrome

The document attached has been archived with permission from the editor of the Medical Journal of Australia. An external link to the publisher’s copy is included.Polycystic ovary syndrome (PCOS) affects 5-20% of women of reproductive age worldwide. The condition is characterized by hyperandrogenism, ovulatory dysfunction and polycystic ovarian morphology (PCOM) - with excessive androgen production by the ovaries being a key feature of PCOS. Metabolic dysfunction characterized by insulin resistance and compensatory hyperinsulinaemia is evident in the vast majority of affected individuals. PCOS increases the risk for type 2 diabetes mellitus, gestational diabetes and other pregnancy-related complications, venous thromboembolism, cerebrovascular and cardiovascular events and endometrial cancer. PCOS is a diagnosis of exclusion, based primarily on the presence of hyperandrogenism, ovulatory dysfunction and PCOM. Treatment should be tailored to the complaints and needs of the patient and involves targeting metabolic abnormalities through lifestyle changes, medication and potentially surgery for the prevention and management of excess weight, androgen suppression and/or blockade, endometrial protection, reproductive therapy and the detection and treatment of psychological features. This Primer summarizes the current state of knowledge regarding the epidemiology, mechanisms and pathophysiology, diagnosis, screening and prevention, management and future investigational directions of the disorder.Robert J Norman, Ruijin Wu and Marcin T Stankiewic

State of the Art Review: Emerging Therapies: The Use of Insulin Sensitizers in the Treatment of Adolescents with Polycystic Ovary Syndrome (PCOS)

PCOS, a heterogeneous disorder characterized by cystic ovarian morphology, androgen excess, and/or irregular periods, emerges during or shortly after puberty. Peri- and post-pubertal obesity, insulin resistance and consequent hyperinsulinemia are highly prevalent co-morbidities of PCOS and promote an ongoing state of excess androgen. Given the relationship of insulin to androgen excess, reduction of insulin secretion and/or improvement of its action at target tissues offer the possibility of improving the physical stigmata of androgen excess by correction of the reproductive dysfunction and preventing metabolic derangements from becoming entrenched. While lifestyle changes that concentrate on behavioral, dietary and exercise regimens should be considered as first line therapy for weight reduction and normalization of insulin levels in adolescents with PCOS, several therapeutic options are available and in wide use, including oral contraceptives, metformin, thiazolidenediones and spironolactone. Overwhelmingly, the data on the safety and efficacy of these medications derive from the adult PCOS literature. Despite the paucity of randomized control trials to adequately evaluate these modalities in adolescents, their use, particularly that of metformin, has gained popularity in the pediatric endocrine community. In this article, we present an overview of the use of insulin sensitizing medications in PCOS and review both the adult and (where available) adolescent literature, focusing specifically on the use of metformin in both mono- and combination therapy

Detection of Coxiella burnetii using (q)PCR: a comparison of available assays

Q fever, caused by the bacterium Coxiella burnetii, has become an emerging public health problem in the Netherlands since 2007. Diagnosis of Q fever, both in humans and animals, is mainly based on serology. Serological techniques are less suitable for direct transmission and source-finding studies for C. burnetii infection due to the delayed detection window for serological tests. The last two decades, several PCR based diagnostic assays (conventional PCR or qPCR) have been developed for the detection of C. burnetii DNA. These assays have been applied for the detection of C. burnetii DNA in clinical samples, veterinary samples, and environmental samples. These PCR-based diagnostic tests are often "in-house" developed assays. A number of commercial PCR diagnostic tests, however, have also become available. A drawback of these commercial kits is that information on some of the components is patented. This makes a thorough assessment of assay performance in relation to other "inhouse" developed assays difficult. In the current study, we describe a comparison of various (q)PCR assays used by laboratories both nationally and internationally for the detection of C. burnetii DNA. We compare the results obtained from three ring trials, set up specifically for the detection of C. burnetii DNA in human, veterinary, or environmental matrices. In addition, we compare specific parameters, such as sensitivity, specificity, and reproducibility for each of the (q)PCR assays. In conclusion, most (q)PCR assays developed for C. burnetii include detection of the multicopy insertion element IS1111, in combination with detection of other single copy genes such as icd, com1, sod, or plasmid genes. PCR based detection assays (conventional PCR or qPCR) for C. burnetii DNA preferably target short and multiple target sequences, including an internal process control in multiplex format. Other performance characteristics have not yet been published for these qPCR assays for the detection of C. burnetii, although this would be recommended.VW